Annual Report. Instituto Carlos III MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD GOBIERNO DE ESPAÑA

Transcription

1 2014 Annual Report GOBIERNO DE ESPAÑA MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD Instituto de Salud Carlos III

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3 Annual Report 2014 Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annua Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annua Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annua Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annua Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annua Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annual Report Annua

4 2015. CIBERNED Centro Investigación Biomédica en Red, Enfermedades Neurodegenerativas C/ Valderrebollo, Madrid Coordination and management of content: Miguel Medina Padilla Jose de Arriba-Enríquez Aina Frontera Sánchez Design and Editorial coordination: Comunicación y Ediciones Sanitarias, S.L. (CYESAN)

5 Index Letter from the Scientific Director... 1 Overview... 2 Aims... 4 Directory of Research Groups And Associated Institutions... 5 Geographic Distribution of CIBERNED Research Groups... 7 Organizational Structure... 8 Organization Chart... 9 External Scientific Advisory Committee Consortium Members Scientific Report Research Programs Program Program Program Cooperative Research International Relations Scientific Productivity and Other Activities Financial Report Principal Investigator Index

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7 2014 Annual Report Letter from the Scientific Director During the second half of this year CIBERNED has undertaken a reorganization of its scientific program structure, so that at the end of 2014, the 54 groups that make up our Center have been grouped together into two programs. Program 1 is called Alzheimer s disease and other dementias and consists of 22 groups, whereas the Program 2 Parkinson s disease and other neurodegenerative motor disorders is composed of the 32 remaining groups. CIBERNED scientific results during this period show the rewards from the projects initiated 4-5 years ago. 588 scientific articles have been published, with an overall accumulates impact of 2,869.2 and an average impact factor of 5.39 per article. There have been four clinical guidelines published and 17 active clinical trials during the year. This indicates our path to consolidate its true nature as a Translational Research Center. More specifically, our center is the one who has published more papers in high impact journals from the Nature, Cell or Science groups, in basic research, or the Lancet or New England Journal of Medicine groups in more clinical research. Several of these publications have been the result of the interaction among CIBERNED groups through the cooperative research program of our Center. Fortunately, these collaborative projects are being efficient and facilitate intramural collaboration, communication and synergy of most of our groups. Additionally, we have continued to collaborate with other groups from other CIBER, inside and outside the InterCIBER program from the Institute of Health Carlos III, the institution on which we depend. Along with the CIEN Foundation and the Queen Sofia Foundation, Institutions with which we collaborate intensively and in the case of CIEN Foundation, shares management, we have carried out the 8 th CIBERNED Scientific Forum, which has been integrated into the Second International Research Conference and Innovation in Neurodegenerative Diseases (CIIIEN). This event took place last September in Barcelona, chaired by HM Queen Sofia, along with the CIBERNED Social Forum, with the participation of different associations of patients with neurodegenerative diseases, mainly from Catalonia. The next edition of CIIIEN, that will include the 9 th CIBERNED Scientific Forum will be held in September 2015, in Malaga. We have continued our international relations, maintaining our active participation in JPND and CoEN activities and projects. It should be noted that our Deputy Director, Dr. Medina, has been appointed to the Ethics Advisory Committee of the Human Brain Project. However, the greatest asset of our Centre is constituted by our researchers who, as already mentioned, have positioned CIBERNED at the head of the Spanish research centers with the highest impact publications. Currently, our Center is very competitive at international level, the level where we have to participate. On the other hand, we hope that with the incipient economic recovery, we can expand our resources and we can accommodate in CIBERNED new research groups of worth, because our intention is making a progressive, but orderly growth, comparable to that performed by good, leading international Centers in our area. We intend that this progressive development will essentially facilitate translational research as a way to improve the lives of those people suffering from neurodegenerative problems in Spain, as this is our ultimate goal. Jesús Ávila de Grado Scientific Director 1

8 Overview The Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED) emerges in 2006 as heir to the Center for Research in Neurological Diseases (CIEN), which together with the CNIO, CNIC or CNMVIS constituted the four centers whose mission is to combat the most prevalent health problems of Spanish society: cancer, cardiovascular diseases, infectious diseases and neurodegeneration. As for the other Centers, a supporting foundation called CIEN Foundation was created in 2003 as the CIEN s management authority, although unlike the other Centers, it was created as the first Network Center in Spain, eventually being renamed as CIBERNED. Both CIBERNED and CIEN Foundation partnered with the Queen Sofia Foundation in creating the Alzheimer Center, located in Vallecas (Madrid) and where CIEN Foundation is headquartered since CIBERNED was founded under the auspices of the Institute of Health Carlos III, under a cooperation agreement signed by the Government and other participating institutions with the idea of creating a Research Center in which basic, clinical and population-based studies are integrated in order to develop a single joint research program, focusing on certain pathologies of great importance for the National Health System either for its prevalence or because of its social impact. It has a multidisciplinary and multiinstitutional character and it is aimed at boosting the impact of cutting research in neurodegenerative disorders through a Network Research Structure, thus contributing to scientifically substantiate the programs and policies of the National Health System in the priority areas of the National R+D+I. CIBERNED is a research organism, endowed with legal status under the Article 6.5 of Law 30/1992 of November 26 on the Legal Regime of Public Administrations and Common Administrative Procedure. It is formed by the association of research groups, with no physical contiguity, belonging to different administrations, institutions and regional governments, from public and private sectors with research lines and goals focused on the specific common area of neurodegenerative diseases and being coordinated to achieve scientific objectives that could hardly be considered in a more restricted execution context. CIBERNED is governed in the internal operating rules, by way of a Regulation. Currently, the CIBERNED-CIEN Foundation partnership is the only research center in Spain (and one of the few in the world) integrated into the international network of Centres of Excellence in Neurodegeneration (CoEN), an initiative arising from the European Union Joint Programme for Research in Neurodegenerative Diseases (JPND). This joint program constitutes an innovative collaborative research initiative created to address the growing challenges posed by this group of diseases. Its goal is to boost the impact of research by aligning existing national research programs and identifying the common objectives whose scope would benefit through joint action. CIBERNED is governed by its own statutes and its activity is currently structured round three Scientific Programs: Program 1: Alzheimer s disease and other degenerative dementias. Program 2: Parkinson s disease, Huntington s chorea and other movement disorders. Program 3: Neuromuscular Diseases. These three Scientific Programs around which the CIBERNED scientific structure was articulated, have become two since the end of The reason for this reduction in the number of programs is due to the voluntary departure of two former Program 3 (Neuromuscular Diseases) groups. Because this exit left at 6 the number of groups in the program, compared to 22 and 26 groups of each of the remaining programs, the CIBERNED Steering Committee decided to request voluntary relocation of these groups in any of the other two programs. All 6 groups considered inclusion in Program 2 as the best option, with the suggestion of a name change for better integration and representation ( Parkinson s disease and other neurodegenerative disorders engine ). This title change would allow the ALS (the third neurodegenerative disease by incidence, after Alzheimer s and Parkinson s) and other neuromuscular diseases (with a cumulative incidence also relevant), be reflected in the CIBERNED scientific program structure. In addition, the Steering Committee decided to launch a reflection period open to all CIBERNED 2

9 2014 Annual Report researchers to discuss whether to establish a new CIBERNED organizational structure, more appropriate to the current situation. Thus, since late 2014, CIBERNED structure is organized around these two main Science Programs: Program 1: Alzheimer s disease and other degenerative dementias. Program 2: Parkinson s disease and other neurodegenerative motor disorders. CIBERNED currently consists of 54 research groups supported by different universities, hospitals and the Research Council (CSIC), each led by a principal investigator or responsible. CIBERNED is a network research center, composed of research groups belonging to different Administrations and affiliated Institutions: researchers members physically work in their parent institutions they belong to while simultaneously and actively participating on CIBERNED s own cooperative research agenda. Thus, is the result of a collaborative partnership between different institutions and the sum of 54 research groups. Depending on the results of the scientific evaluation of the various research groups, there is the possibility to agree on the discontinuation of some groups and, depending on budget availability, it may also consider adding some new research group currently outside CIBERNED, but that satisfied the requirements of scientific quality and translational activity and it is aligned with CIBERNED priorities. The Scientific Director Office has been set since October 2011 at the Center for Molecular Biology Severo Ochoa (CSIC) in Madrid. The headquarters, including the General Manager Office is located in the Alzheimer Center of the Queen Sofia Foundation, 5 Valderrebollo street in Madrid. 3

10 Aims CIBERNED s ultimate goal is to foster scientific and technical research of excellence in the field of human health, with the overall goal of producing results quickly translatable into clinical practice to improve the health and wellbeing of patients with neurodegenerative diseases, as well as their families and caregivers, providing in turn economic and social benefits. Among its objectives the following should be highlighted: Promote and develop cooperative translational research excellence in neurodegenerative diseases. Fostering the impact of science on the health system and on the welfare of patients. Enhance participation in coordinated actions and calls promoted by funding agencies and the international and national level. Encourage the development of new therapies. Implement an infrastructure of strategic value for the development of research on prevention, diagnosis and treatment. Develop plans for specialized training. Promote support cross-platforms (biobanks, brain imaging, etc). To involve society in the enormous medical and socioeconomic impact of neurodegenerative diseases and facilitate their involvement in the fight against neurodegeneration. 4

11 2014 Annual Report Directory of Research Groups and Associated Institutions Group Principal Investigator Institution 301 Alberch Vié, Jordi University of Barcelona 401 Ávila de Grado, Jesús Center for Molecular Biology "Severo Ochoa" CSIC-UAM, Madrid 601 Berciano Blanco, José Ángel Marqués de Valdecilla Foundation, Santander 502 Bermejo Pareja, Félix 12 de Octubre University Hospital, Madrid 510 Bullido Gómez-Heras, María Jesús Autonomous University, Madrid 402 Camins Espuny, Antonio University of Barcelona 201 Canela Campos, Enric Isidre University of Barcelona 511 Cantero Lorente, José Luis Pablo de Olavide University, Sevilla 106 Ceña Callejo, Valentín University of Castilla-La Mancha, Albacete 413 Comella Carnicé, Joan Xavier Vall d'hebron University Hospital, Barcelona 101 Cuadrado Pastor, Antonio Autonomous University of Madrid 403 de Felipe Oroquieta, Javier Cajal Institute CSIC; Technical University, Madrid 509 de Pedro Cuesta, Jesús Institute of Health Carlos III, Madrid 114 del Río Fernández, José Antonio Catalonian Institute de Bioengineering, Barcelona 102 Fariñas Gómez, Isabel University of Valencia 606 Fernández Chacón, Rafael University of Seville 303 Fernández Ruiz, Javier Complutense University of Madrid 503 Ferrer Abizanda, Isidro Bellvitge Institute of Biomedical Research, Barcelona 103 Fuentes Rodríguez, José Manuel University of Extremadura, Cáceres 113 García Verdugo, José Manuel Cavanilles Institute, University of Valencia 104 González Castaño, José Autonomous University of Madrid 415 Gutiérrez Pérez, Antonia University of Málaga 305 Guzmán Pastor, Manuel Complutense University of Madrid 111 Iglesias Vacas, Teresa Institute of Biomedical Research CSIC-UAM, Madrid 603 Illa Sendra, Isabel * Santa Creu i Sant Pau Hospital, Barcelona 202 Kulisevsky Bojarski, Jaime Santa Creu i Sant Pau Hospital, Barcelona 208 Labandeira García, José Luis University of Santiago de Compostela 203 Lanciego Pérez, José Luis Center of Applied Medical Research, Univ. Navarra, Pamplona 504 Lleó Bisa, Alberto Santa Creu i Sant Pau Hospital, Barcelona 105 López Barneo, José Virgen del Rocío University Hospital, University of Sevilla 506 López de Ceballos Lafarga, María Cajal Institute CSIC, Madrid 609 López de Munain Arregui, Adolfo Biodonostia Research Institute 5

12 Group Principal Investigator Institution 306 Lucas Lozano, José Javier Center for Molecular Biology "Severo Ochoa" CSIC-UAM, Madrid 404 Matute Almau, Carlos University of the Basque Country, Bilbao 304 Mena Gómez, María Ángeles ** Ramón y Cajal University Hospital, Madrid 508 Mengod Los Arcos, Guadalupe Institute of Biomedical Research IDIBAPS-CSIC, Barcelona 204 Moratalla Villalba, Rosario Cajal Institute CSIC, Madrid 604 Muñoz Cánoves, Pura Pompeu Fabra University, ICREA, Barcelona 307 Naranjo Orovio, José Ramón National Center of Biotechnology, CSIC, Madrid 607 Navarro Acebes, Xavier Autonomous University of Barcelona 205 Obeso Inchausti, José Ángel *** Center of Applied Medical Research, Univ. Navarra, Pamplona 507 Pastor Muñoz, María Asunción Center of Applied Medical Research, Univ. Navarra, Pamplona 110 Pérez Castillo, Ana María Institute of Biomedical Research CSIC-UAM, Madrid 209 Pérez Tur, Jordi Institute of Biomedicine of Valencia, CSIC 406 Rodríguez Álvarez, José Autonomous University of Barcelona 206 Rodríguez Díaz, Manuel University of La Laguna, Tenerife 407 Sáez Valero, Javier Miguel Hernández University, Elche 408 Soriano García, Eduardo University of Barcelona 207 Tolosa Sarró, Eduardo Hospital Clinic of Barcelona 409 Torres Alemán, Ignacio Cajal Institute CSIC, Madrid 410 Trullás Oliva, Ramón Institute of Biomedical Research IDIBAPS-CSIC, Barcelona 108 Vicario Abejón, Carlos Cajal Institute CSIC, Madrid 109 Vila Bover, Miquel Vall d'hebron University Hospital, Barcelona 605 Vílchez Padilla, Juan Jesús * La Fe University Hospital, Valencia 411 Vitorica Ferrández, Francisco Javier University of Seville 412 Wandosell Jurado, Francisco Center for Molecular Biology "Severo Ochoa" CSIC-UAM, Madrid * Volunteer transfer to CIBERER (June 2014) ** Takes over from Justo García de Yébenes Prous due to retirement *** Hospital de Madrid Foundation (from October 2014) 6

13 2014 Annual Report Geographic Distribution of CIBERNED Research Groups

14 Organizational Structure CIBERNED governing bodies are the Governing Council, the Permanent Commission and the Scientific Director. The Governing Council, the highest body of CIBERNED, consists of three representatives of the Institute of Health Carlos III, that include the legal representative of the collaborating organization, the Foundation for Research in Neurological Diseases (CIEN) and one representative from each of the institutions participating in the consortium. The President of the Executive Council is the Director of the Institute of Health Carlos III. The Secretary of the Governing Council will be the Manager of the consortium The Scientific Director and Manager of CIBERNED are also part of the Governing Council, without a right to vote. The Standing Committee is composed of the Vice President of the Executive Council or his delegate, and four members representing the consortium institutions in the Governing Council. The Scientific Director of the consortium and the Manager, who acts as secretary, are also part of the Permanent Commission, without a right to vote. The Scientific Director is appointed by the President of the Governing Council for a period of four years, renewable by agreement of the parties. The current Scientific Director of CIBERNED is Dr. Jesús Ávila de Grado. CIBERNED Management is entrusted to the Managing Director of the Foundation CIEN, Ms. María Ángeles Pérez Muñoz. Support and advisory bodies to the organs of government: a. The Steering Committee b. The External Scientific Advisory Committee The Steering Committee consists of: Dr. Jesús Ávila de Grado Scientific Director Ms. María Ángeles Pérez Muñoz Manager Dr. Miguel Medina Padilla Deputy Scientific Director and the Program coordinators: Dr. Isidro Ferrer Abizanda Dr. Albert Lleó Bisa Dr. Adolfo López de Munain Dr. José Javier Lucas Lozano Dr. Eduardo Soriano García Dr. Eduardo Tolosa Sarró 8

15 2014 Annual Report Ciberned Organization Chart Institute of Health Carlos III Governing Council Associated Institutions Standing Committee Steering Committee Deputy Scientific Director Scientific Director Research Program Coordinators Manager Research Groups 9

16 External Scientific Advisory Committee Dr. George Perry Dr. Vincenzo Bonifati Dr. Angel Cedazo-Mínguez Dr. Mary Reilly University of Texas, San Antonio, USA (Chairman) Erasmus University, Rotterdam, The Netherlands Karolinska Institute, Stockholm, Sweden National Hospital for Neurology and Neurosurgery and Institute of Neurology, London, United Kingdom 10

17 2014 Annual Report Consortium Members The Central Government represented by: The Institute of Health Carlos III. The Spanish National Research Council (CSIC). The following Autonomous Regions (Comunidades Autónomas): Autonomous Region of Andalusia, through the Andalusian Public Foundation for Health Research Management at Seville, University of Seville, Pablo de Olavide University, and University of Málaga. Autonomous Region of Canarias, through the University of La Laguna. Autonomous Region of Cantabria, through the Marqués de Valdecilla Foundation - IDIVAL. Autonomous Region of Castilla-La Mancha, through the University of Castilla-La Mancha. Autonomous Region of Catalonia, through the Pompeu Fabra University, Research Institute Vall d Hebron Foundation, Bellvitge Biomedical Research Institute Foundation, University of Barcelona and Autonomous University of Barcelona. Autonomous Region of Valencia, through La Fe Hospital Research Foundation, Príncipe Felipe Research Centre, University of Valencia and Miguel Hernández University at Elche. Autonomous Region of Extremadura, through the Foundation for Research and Training of Health Professionals (FUNDESALUD). Autonomous Region of Madrid, through Madrid Health Services (12 de Octubre University Hospital and Ramón y Cajal Hospital), Autonomous University of Madrid and Complutense University of Madrid. Autonomous Region of the Basque Country, through the University of the Basque Country and Biodonostia Research Institute. Autonomous Region of Galicia, through the University of Santiago de Compostela. Other centers and institutions that are not affiliated to the previously-cited public administrations: Santa Creu i Sant Pau Hospital Biomedical Research Institute Foundation. Center for Applied Medical Research (CIMA). Hospital Clinic of Barcelona. 11

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21 2014 Annual Report Research Programs The structure of CIBERNED research groups is organized through three scientific programs, whose goals, composition and structure is maintained as in previous years, and whose budget allocation is based on the scientific results of the internal scientific evaluation. The distribution of these three thematic programs shows CIBERNED commitment to encouraging translational research and obtaining scientific knowledge resulting in improving the prevention, diagnosis and treatment of neurodegenerative diseases. All this without undermining the development of transversal research projects of interest to different areas (see below). The aim of the structure described is to provide a coherent conceptual scheme embodying the real cooperative scientific activity conducting various research groups. As described in the Overview section, these three scientific programs around which the CIBERNED scientific structure has been articulated in recent years, have become two by the end of The reason for this reduction in the number of programs is a consequence of the voluntary departure of two groups from the former program 3 (Neuromuscular Diseases), so that the CIBERNED Steering Committee decided to merge Programs 2 and 3, with the suggestion of a name change for better integration and representation ( Parkinson s disease and other neurodegenerative motor disorders ). In this way, amyotrophic lateral sclerosis (ALS, the third neurodegenerative disease by incidence, after Alzheimer s and Parkinson s) and other neuromuscular diseases (also with a significant cumulative incidence) are reflected in the CIBERNED scientific program structure. The Programs are described below. 15

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23 Alzheimer s Disease and other Degenerative Dementias Program 1 Jesús Ávila de Grado Group Félix Bermejo Pareja Group María Jesús Bullido Gómez-Heras Group Antonio Camins Espun Group José Luis Cantero Lorente Group Joan Xavier Comella Carnicé Group Javier de Felipe Oroquieta Group Jesús d Pedro Cuesta Group Isidro Ferrer Abizanda Group Antonia Gutiérrez Pérez Group Alberto Lleó Bisa Group María Lópe de Ceballos Lafarga Group Carlos Matute Almau Group Guadalupe Mengod Los Arcos Group María Asunción Pasto Muñoz Group José Rodríguez Álvarez Group Javier Sáez Valero Group Eduardo Soriano García Group Ignacio Torre Alemán Group Ramón Trullás Oliva Group Francisco Javier Vitorica Ferrández Group Francisco Wandosell Jurado Group Jesús Ávila de Grado Group Félix Bermejo Pareja Group María Jesús Bullido Gómez-Heras Group Antonio Camins Espun Group José Luis Cantero Lorente Group Joan Xavier Comella Carnicé Group Javier de Felipe Oroquieta Group Jesús d Pedro Cuesta Group Isidro Ferrer Abizanda Group Antonia Gutiérrez Pérez Group Alberto Lleó Bisa Group María Lópe de Ceballos Lafarga Group Carlos Matute Almau Group Guadalupe Mengod Los Arcos Group María Asunción Pasto Muñoz Group José Rodríguez Álvarez Group Javier Sáez Valero Group Eduardo Soriano García Group Ignacio Torre Alemán Group Ramón Trullás Oliva Group Francisco Javier Vitorica Ferrández Group Francisco Wandosell Jurado Group Jesús Ávila de Grado Group Félix Bermejo Pareja Group María Jesús Bullido Gómez-Heras Group Antonio Camins Espun Group José Luis Cantero Lorente Group Joan Xavier Comella Carnicé Group Javier de Felipe Oroquieta Group Jesú de Pedro Cuesta Group Isidro Ferrer Abizanda Group Antonia Gutiérrez Pérez Group Alberto Lleó Bisa Group Marí López de Ceballos Lafarga Group Carlos Matute Almau Group Guadalupe Mengod Los Arcos Group María Asunció Pastor Muñoz Group José Rodríguez Álvarez Group Javier Sáez Valero Group Eduardo Soriano García Group Ignaci Torres Alemán Group Ramón Trullás Oliva Group Francisco Javier Vitorica Ferrández Group Francisco Wandosell Jurado Research Groups Ávila de Grado, Jesús Bermejo Pareja, Félix Bullido Gómez-Heras, María Jesús Camins Espuny, Antonio Cantero Lorente, José Luis Comella Carnicé, Joan Xavier de Felipe Oroquieta, Javier de Pedro Cuesta, Jesús Ferrer Abizanda, Isidro Gutiérrez Pérez, Antonia Lleó Bisa, Alberto López de Ceballos Lafarga, María Matute Almau, Carlos Mengod Los Arcos, Guadalupe Pastor Muñoz, María Asunción Rodríguez Álvarez, José Sáez Valero, Javier Soriano García, Eduardo Torres Alemán, Ignacio Trullás Oliva, Ramón Vitorica Ferrández, Francisco Javier Wandosell Jurado, Francisco

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25 2014 Annual Report Scientific Report Program 1 Increased life expectancy is a reflection of the economic, social and health progress achieved in developed countries during the last century. However, the aging of the population carries some new scientific and public health challenges by increasing the appearance of old age-specific diseases. Dementia is nowadays one of the degenerative conditions more frequently observed in this population. According to OECD estimates, about 15% of those aged between 80 and 84 years, and nearly one in four of those over 85 years suffer from some form of dementia (OECD, 2011). According to WHO, about 35.6 million people are living with dementia. With the aging population and without effective prevention options, this number is expected to double by 2030 and more than triple by 2050 (WHO and Alzheimer s Disease International, 2012). Alzheimer s disease (AD) is the most common form of dementia. It is a syndrome that can result from a series of progressive disorders affecting memory processes, cognition, behavior and ability to perform daily living activities. AD is a relatively poorly understood disorder with few effective treatments and high, increasing economic and social consequences. The disorder primarily affects the elderly, although there is a growing awareness of cases that start before age 65 in many developed countries, is the leading cause of disability in people over 65, and one of the major causes of death after stroke and cardiovascular disease. Although most individuals with AD are no longer within working age, the disease poses a significant economic burden due to lost productivity, mainly in the form of unpaid care and social care. Despite advances in our understanding of the neurobiology of AD, we still have much to learn regarding the molecular basis underlying the deficit of neural systems, which manifests as brain dysfunction or behavior alterations, including dementia. We need a deeper understanding of these biological mechanisms for the development of effective treatments and directed them to slow down or alter the disease progression. AD is a complex and multifactorial disease, which implies the existence of very few, and only partially effective, drugs accessible to patients. Diagnosis of the disease is still difficult, particularly when it is at an early stage. Find only partially effective new disease markers and related therapeutic targets is a critical challenge. AD is characterized by the presence of two aberrant structures within the patients brains, senile plaques and neurofibrillary tangles, synapse loss (it is considered a synaptopathy), especially among hippocampal and cortical neurons, as well as a considerable neurodegeneration. Some aspects of these pathologies may be reproduced in cellular or animal models. At present, dozens of laboratories around the world are actively working to identify new causative and risk genes involved in these pathologies, which could contribute to clarify its pathophysiological basis and identify new therapeutic targets. One of the main current issues in AD is that when diagnosed the brain has already suffered such an extensive and irreparable damage, so that finding biomarkers for an earlier detection becomes an urgent need, even at an asymptomatic stage, when any therapeutic strategy would have a greater chance of success. Advances in genomics and other omics technologies are opening opportunities to discover new diagnostic and therapeutic targets. For instance, advances in sequencing technologies now make it possible to perform association studies and sequencing of the entire genome on a large scale. Comparing data between culturally or genetically different cohorts, as donors of representative samples of genetic material or otherwise, can provide an essential insight into the pathogenic mechanisms underlying the disease and help to identify potential therapeutic targets. Such information is essential for the discovery of potential biomarkers. It is also of interest to get a more accurate risk assessment leading to a more accurate and timely prediction of people at risk of developing the disease, including identifying potential environmental modifiers of disease risk. There are also other technologies such as regenerative medicine, nanotechnology and gene therapy that could, in the medium or long term, provide innovative solutions to deal with the complex pathogenic mechanisms of the disease. 19

26 Within this program, 22 groups of clinical and basic scientists, engaged on Alzheimer s diagnosis and care of patients as well as on basic research in the laboratory, combine their expertise and effort to work in a coordinated way in the search for new genetic factors, disease-related biomarkers and new therapeutic strategies for AD and other degenerative dementias. The main research lines are the following: Genetic Epidemiology Research on disease-related biomarkers Cellular and animal models of Alzheimer s disease and other degenerative dementias Molecular pathology of Alzheimer s disease Mechanisms of neurodegeneration, neuroprotection and design of new therapies. Program 1 20

27 2014 Annual Report Scientific Report Groups Principal Investigator Ávila de Grado, Jesús Bermejo Pareja, Félix Bullido Gómez-Heras, María Jesús Camins Espuny, Antonio Cantero Lorente, José Luis Comella Carnicé, Joan Xavier de Felipe Oroquieta, Javier de Pedro Cuesta, Jesús Ferrer Abizanda, Isidro Gutiérrez Pérez, Antonia Lleó Bisa, Alberto López de Ceballos Lafarga, María Matute Almau, Carlos Mengod Los Arcos, Guadalupe Pastor Muñoz, María Asunción Rodríguez Álvarez, José Sáez Valero, Javier Soriano García, Eduardo Torres Alemán, Ignacio Trullás Oliva, Ramón Vitorica Ferrández, Francisco Javier Wandosell Jurado, Francisco Institution Center of Molecular Biology Severo Ochoa (CSIC-UAM), Madrid 12 de Octubre Hospital, Madrid Center of Molecular Biology Severo Ochoa, Autonomous University of Madrid University of Barcelona Pablo de Olavide University, Seville Foundation Vall d Hebron University Hospital, Research Institute, Barcelona Cajal Institute, CSIC / Center of Biomedical Technology, Technical University of Madrid Institute of Health Carlos III, Madrid IDIBELL, Bellvitge University Hospital / University of Barcelona University of Málaga Santa Creu i Sant Pau Hospital, Barcelona Cajal Institute, CSIC, Madrid University of the Basque Country, Bilbao CSIC, IDIBAPS, Barcelona Foundation of Applied Medical Research, University of Navarra, Pamplona Institute of Neuroscience, Autonomous University of Barcelona Miguel Hernández University, Elche University of Barcelona Cajal Institute, CSIC, Madrid CSIC, IDIBAPS, Barcelona University of Seville Center of Molecular Biology Severo Ochoa (CSIC-UAM), Madrid Program 1 is coordinated by Drs. Isidro Ferrer (IDIBELL - Bellvitge Hospital), Alberto Lleó (Santa Creu i San Pau Hospital) and Eduardo Soriano (University of Barcelona). 21

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29 Group Jesús Ávila de Grado Program 1 CONTACT DETAILS Jesús Ávila de Grado Group Jesús Ávila de Grado Group Jesús Centro Ávila de de Biología Grado Group Molecular Jesús Severo Ávila Ochoa de Grado Group Jesú Ávila de Grado Group Jesús Ávila de Grado Group Jesús Ávila de Grado C/ Nicolás Group Cabrera, Jesús 1 Ávila (Laboratorio de Grado 208) Group Jesús Ávil de Grado Group Jesús Ávila de Grado Group Jesús Ávila de Grado Group Jesús Campus Ávila de de Grado Cantoblanco Group Jesús Ávila d Madrid (Spain) Grado Group Jesús Ávila de Grado Group Jesús Ávila de Grado Group Jesús Ávila Phone: de Grado Group 964 Jesús 564 Ávila de Grad Group Jesús Ávila de Grado Group Jesús Ávila de Grado Group Jesús Ávila de Grado Fax: +34 Group Jesús 420Ávil a de Ávila d javila@cbm.csic.es Grado Group Jesús Ávila de Grado Group Jesús Ávila de Grado Group Jesús Ávila de Grado Ávila de Grado Group Jesú LIST OF PERSONNEL Jesús Ávila de Grado Principal Investigator, Research Professor Félix Hernández Pérez Researcher María Llorens-Martín Researcher Vega García-Escudero Researcher Alberto Gómez Ramos Researcher Almudena Fuster Matanzo Researcher Jerónimo Jurado Arjona Researcher Patricia Martín-Maestro PhD student Noemí Pallas Bazarra PhD student Jesús Merchán Robira PhD student Raquel Cuadros Catalán Technician Esther García García Technician Nuria de la Torre Alonso Administrative Assistant 23

30 Summary It has been indicated a deficit in the function of adult neurogenesis, occurring at dentate gyrus, that can take place in demented patients. To look at this feature, we have used a mouse model having an impaired adult neurogenesis. This analysis has complemented a previous work and, in this year, we have focused on the influence of inflammation on that process. Also, we have used anti-inflammatory compounds to look for a possible therapeutically approach to reverse the inflammatory effect on adult neurogenesis. About tau, we have studied the role of the protein on axonogenesis. Our results have indicated that tau plays an important role at the last step of axonogenesis in a process in which changes in the location of EB1/EB3 on microtubules take place. During this time we have collaborated with other groups of CIBERNED, mainly with that of Dr. Soriano. Our collaborative work has been related to the search of specific somatic mutations in brain cells from Alzheimer disease patients. Most recently, we have collaborated with Dr. Lucas in a work on Huntington disease mainly developed by his group. Keywords Tau, GSK3, Alzheimer disease, Neurogenesis Publications El Kadmiri N, Cuardos R, El Moutawakil B, Slassi I, Avila J, Nadifi S, et al. A Proteomic Approach for the Involvement of the GAPDH in Alzheimer Disease in the Blood of Moroccan FAD Cases. Journal of molecular neuroscience: MN. 2014;54(4): Epub 2014 Jul 15. Rabano A, Rodal I, Cuadros R, Calero M, Hernandez F, Avila J. Argyrophilic grain pathology as a natural model of tau propagation. Journal of Alzheimer s disease: JAD. 2014;40(Suppl 1):S Perez M, Cuadros R, Pallas-Bazarra N, Garcia C, Langa E, Jurado-Arjona J, et al. Boronate-tau mediated uptake in neurons. Journal of Alzheimer s disease: JAD. 2014;40(1): Sayas CL, Avila J. Crosstalk between axonal classical microtubule-associated proteins and end binding proteins during axon extension: possible implications in neurodegeneration. Journal of Alzheimer s disease: JAD. 2014;40(Suppl 1):S Llorens-Martin M, Jurado J, Hernandez F, Avila J. GSK-3β, a pivotal kinase in Alzheimer disease. Frontiers in molecular neuroscience. 2014;7:46. Epub 2014 May 21. Medina M, Avila J. Is tau a prion-like protein? Journal of Alzheimer s disease: JAD. 2014;40(Suppl 1):S1-3. Barbosa DJ, Serrat R, Mirra S, Quevedo M, Gomez de Barreda E, Avila J, et al. MDMA impairs mitochondrial neuronal trafficking in a Tau- and Mitofusin2/Drp1-dependent manner. Archives of toxicology. 2014;88(8): Epub 2014 Feb 13. Medina M, Avila J. New perspectives on the role of tau in Alzheimer s disease. Implications for therapy. Biochemical pharmacology. 2014;88(4): Epub 2014 Jan 22. Llorens-Martin M, Jurado-Arjona J, Avila J, Hernandez F. Novel connection between newborn granule neurons and the hippocampal CA2 field. Experimental neurology. 2015;263: Epub 2014 Nov 1. Llorens-Martin M, Jurado-Arjona J, Fuster-Matanzo A, Hernandez F, Rabano A, Avila J. Peripherally triggered and GSK-3β-driven brain inflammation differentially skew adult hippocampal neurogenesis, Program 1 24

31 2014 Annual Report Jesús Ávila de Grado Group behavioral pattern separation and microglial activation in response to ibuprofen. Translational psychiatry. 2014;4:e463. Epub 2014 Oct 14. Sayas CL, Avila J. Regulation of EB1/3 proteins by classical MAPs in neurons. Bioarchitecture. 2014;4(1):1-5. Epub 2014 Jan 10. Gomez-Ramos A, Sanchez-Sanchez R, Muhaisen A, Rabano A, Soriano E, Avila J. Similarities and differences between exome sequences found in a variety of tissues from the same individual. PloS one. 2014;9(7):e Epub 2014 Jul 1. Parcerisas A, Rubio SE, Muhaisen A, Gomez-Ramos A, Pujadas L, Puiggros M, et al. Somatic signature of brain-specific single nucleotide variations in sporadic Alzheimer s disease. Journal of Alzheimer s disease: JAD. 2014;42(4): Avila J, Simon D, Diaz-Hernandez M, Pintor J, Hernandez F. Sources of extracellular tau and its signaling. Journal of Alzheimer s disease: JAD. 2014;40(Suppl 1):S7-S15. Camero S, Benitez MJ, Barrantes A, Ayuso JM, Cuadros R, Avila J, et al. Tau protein provides DNA with thermodynamic and structural features which are similar to those found in histone-dna complex. Journal of Alzheimer s disease: JAD. 2014;39(3): Martinez-Aguila A, Fonseca B, Hernandez F, Diaz-Hernandez M, Avila J, Pintor J. Tau triggers tear secretion by interacting with muscarinic acetylcholine receptors in New Zealand white rabbits. Journal of Alzheimer s disease: JAD. 2014;40(Suppl 1):S71-7. Barbosa DJ, Serrat R, Mirra S, Quevedo M, de Barreda EG, Àvila J, et al. The mixture of ecstasy and its metabolites impairs mitochondrial fusion/fission equilibrium and trafficking in hippocampal neurons, at in vivo relevant concentrations. Toxicological sciences: an official journal of the Society of Toxicology. 2014;139(2): Epub 2014 Mar 4. Medina M, Avila J. The need for better AD animal models. Frontiers in pharmacology. 2014;5:227. Epub 2014 Oct 24. Medina M, Avila J. The role of extracellular Tau in the spreading of neurofibrillary pathology. Frontiers in cellular neuroscience. 2014;8:113. Epub 2014 Apr 23. Camero S, Benitez MJ, Cuadros R, Hernandez F, Avila J, Jimenez JS. Thermodynamics of the interaction between Alzheimer s disease related tau protein and DNA. PloS one. 2014;9(8):e Epub 2014 Aug 15. Avila J, Gomez-Ramos A, Soriano E. Variations in brain DNA. Frontiers in aging neuroscience. 2014;6:323. Epub 2014 Nov 25. de Cristobal J, Garcia-Garcia L, Delgado M, Perez M, Pozo MA, Medina M. Longitudinal assessment of a transgenic animal model of tauopathy by FDG-PET imaging. Journal of Alzheimer s disease: JAD. 2014;40(Suppl 1):S Llorens-Martin M, Blazquez-Llorca L, Benavides-Piccione R, Rabano A, Hernandez F, Avila J, et al. Selective alterations of neurons and circuits related to early memory loss in Alzheimer s disease. Front Neuroanat. 2014;8:38. Epub 2014 May 27. Fernandez-Nogales M, Cabrera JR, Santos-Galindo M, Hoozemans JJ, Ferrer I, Rozemuller AJ, et al. Huntington s disease is a four-repeat tauopathy with tau nuclear rods. Nature medicine. 2014;20(8): Epub 2014 Jul 20. de Cristobal J, Garcia-Garcia L, Delgado M, Pozo MA, Medina M. A longitudinal FDG-PET study of transgenic mice overexpressing GSK- 3β in the brain. Current Alzheimer research. 2014;11(2): Medina M, Avila J. New insights into the role of glycogen synthase kinase-3 in Alzheimer s disease. Expert opinion on therapeutic targets. 2014;18(1): Epub 2013 Oct 8. Camero S, Benitez MJ, Barrantes A, Ayuso JM, Cuadros R, Avila J, et al. Tau protein provides DNA with thermodynamic and structural features which are similar to those found in histone-dna complex. Journal of Alzheimer s disease: JAD. 2014;39(3):

32 Research Projects Code: SAF Title: Función de GSK3 y la proteína tau en neurogénesis y neurodgeneración. Las consecuencias en para enfermedad de Alzheimer Principal Investigator: Jesús Ávila de Grado CIBERNED s groups: Type: National Funding Agency: MICINN Funding: Duration: Code: S2010_BMD Title: Redes de señalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas Principal Investigator: José González Castaño CIBERNED s collaboration: Yes CIBERNED s groups: G401; G104; G111; G307; G409 Other CIBER s collaboration: CIBERER Type: International Funding Agency: Comunidad de Madrid Funding: Duration: Code: 2013/07A Title: Papel de GSK-3 β en las alteraciones de los circuitos corticales que ocurren en la enfermedad de Alzheimer Principal Investigator: Teresa Iglesias Vacas CIBERNED s collaboration: Yes CIBERNED s groups: G401; G403; G111; G504; G307 Type: International Funding Agency: CIBERNED Funding: Duration: Program 1 26

33 Group Félix Bermejo Pareja Program 1 CONTACT DETAILS Group Félix Bermejo Pareja Group Félix Bermejo Pareja Instituto Group de Félix Investigación Bermejo Pareja Hospital Group 12 de Félix Octubre Bermejo Pareja Grou Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Avda. Group de Félix Córdoba Bermejo s/n Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Madrid Bermejo (Spain) Pareja Group Fél Phone: Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Be Fax: mejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo carroeva@h12o.es Pareja Group Félix Bermej Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo Pareja Group Félix Bermejo P LIST OF PERSONNEL Félix Bermejo Pareja Collaborator Eva María Carro Díaz Principal Investigator Teresa Díaz San Juan PhD student Agnieszka Krzyzanowska Postdoctoral researcher Consuelo Pascual Pérez Lab Technician Desiree Antequera Tienda Research Technician Fernando Bartolomé Postdoctoral researcher José Antonio Molina Arjona Neurologist, Scientific staff Julián Benito León Neurologist, Scientific staff Jesús Hernández Gallego Neurologist, Scientific staff Rocío Trincado Soriano Stadistics Technician Alberto Villarejo Galende Neurologist, Scientific staff Álvaro Sánchez-Ferro Río Hortega Researcher Ignacio J Posada Neurologist, Scientific staff Fernando Sierra Río Hortega Researcher 27

34 Summary The research group of Dr. Carro in 2014 has focused its researcher work on Neuropathological mechanisms and searching for possible therapeutic targets for Alzheimer s disease. In this context she produced several investigations with transgenic mice APP/Ps1, improving cognitive deficits, and reducing cerebral accumulation of beta-amyloid and other aspects of Alzheimer disease biology. Dr. Félix Bermejo-Pareja (now retired of his post of Head of the Neurology Department and working as Consultant Research in the Imas12 Institute) group has continued the publication of the cohort NEDICES, primarily on mortality of neurological illnesses (dementia and Parkinson s) and other works in this field. In addition, the mortality of systemic diseases is being investigated. Collaborations with Dr. MA Martín-Requero (Centro de Investigaciones Biológicas, CSIC) team, and genetic aspects of Alzheimer disease with research group of the Queen Sofia Institute, also with the Dr. I Contador from Psychology in Salamanca University about cognitive aspect of the NEDICES cohort. Keywords Alzheimer disease, Dementia, Biomarkers, Transgenic mice, Choroid plexus, Therapeutic drugs, Amyloid, Neurogenesis, Neurodegenerative disorders mortality, Mild cognitive impairment Publications Sanchez-Ferro A, Rabano A, Catalan MJ, Rodriguez-Valcarcel FC, Diez SF, Herreros-Rodriguez J, et al. In vivo gastric detection of α-synuclein inclusions in Parkinson s disease. Mov Disord. [Epub ahead of print]. Romero JP, Benito-Leon J, Louis ED, Bermejo-Pareja F. Alzheimer s disease is associated with decreased risk of cancer-specific mortality: a prospective study (NEDICES). Journal of Alzheimer s disease: JAD. 2014;40(2): Dietrich M, Antequera D, Pascual C, Castro N, Bolos M, Carro E. Alzheimer s disease-like impaired cognition in endothelial-specific megalin-null mice. Journal of Alzheimer s disease: JAD. 2014;39(4): Höftberger R, Sepulveda M, Armangue T, Blanco Y, Rostasy K, Cobo Calvo A, et al. Antibodies to MOG and AQP4 in adults with neuromyelitis optica and suspected limited forms of the disease. Multiple sclerosis (Houndmills, Basingstoke, England). [Epub ahead of print]. Epub 2014 Oct 24. Contador I, Bermejo-Pareja F, Mitchell AJ, Trincado R, Villarejo A, Sanchez-Ferro A, et al. Cause of death in mild cognitive impairment: a prospective study (NEDICES). European journal of neurology: the official journal of the European Federation of Neurological Societies. 2014;21(2):253-e9. Epub 2013 Oct 15. Labiano-Fontcuberta A, Mitchell AJ, Moreno-Garcia S, Benito-Leon J. Cognitive impairment in patients with multiple sclerosis predicts worse caregiver s health-related quality of life. Multiple sclerosis (Houndmills, Basingstoke, England). 2014;20(13): Epub 2014 Apr 28. Pozuelo-Moyano B, Benito-Leon J. Diet and multiple sclerosis. Revista de neurologia. 2014;58(10): Ibanez J, Gonzalez de la Aleja J, Gallego JA, Romero JP, Saiz-Diaz RA, Benito-Leon J, et al. Effects of alprazolam on cortical activity and tremors in patients with essential tremor. PloS one. 2014;9(3):e Epub 2014 Mar 25. Program 1 28

35 2014 Annual Report Félix Bermejo Pareja Group Benito-Leon J. Essential tremor: a neurodegenerative disease? Tremor and other hyperkinetic movements (New York, N.Y.). 2014;4:252. Epub 2014 Jul 22. Benito-Leon J, Romero JP, Louis ED, Bermejo-Pareja F. Faster cognitive decline in elders without dementia and decreased risk of cancer mortality: NEDICES Study. Neurology. 2014;82(16): Epub 2014 Apr 9. Leon Ruiz M, Benito-Leon J, Garcia-Soldevilla MA, Arranz Caso JA, Garcia-Albea Ristol E. First described case of coma triggered by retrograde venous air embolism: an exceptional but potentially lethal situation. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Jul 8. Velasco SL, Sierra-Hidalgo F, Rodriguez RM, Guerreo AJ, Morales JR. Flecainide-induced myoclonus. Clinical neuropharmacology. 2014;37(2):65-6. Esteras N, Alquezar C, Bartolome F, de la Encarnacion A, Bermejo-Pareja F, Molina JA, et al. G1/S Cell Cycle Checkpoint Dysfunction in Lymphoblasts from Sporadic Parkinson s Disease Patients. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Sep 3. Fernandez-Ruiz M, Guerra-Vales JM, Trincado R, Medrano MJ, Benito-Leon J, Bermejo-Pareja F. Hip fracture in three elderly populations of central Spain: data from the NEDICES study. Internal and emergency medicine. 2014;9(1): Epub 2011 Nov 23. Labiano-Fontcuberta A, Mitchell AJ, Moreno-Garcia S, Puertas-Martin V, Benito-Leon J. Impact of anger on the health-related quality of life of multiple sclerosis patients. Multiple sclerosis (Houndmills, Basingstoke, England). [Epub ahead of print]. Epub 2014 Nov 28. Sierra-Hidalgo F, Romero JP, Bermejo-Pareja F, Sanchez-Ferro A, Hernandez-Gallego J, Posada IJ, et al. Inter-Rater Agreement in the Clinical Diagnosis of Essential Tremor: Data from the NEDICES-2 Pilot Study. Tremor Other Hyperkinet Mov (N Y). 2014;4. Epub 2014 Feb 4. Guerrero AM, Sierra-Hidalgo F, Calleja P, Navia P, Campollo J, Diaz-Guzman J. Intracranial Internal Carotid Artery Angioplasthy and Stenting in Giant Cell Arteritis. Journal of neuroimaging: official journal of the American Society of Neuroimaging. [Epub ahead of print]. Epub 2014 Apr 7. Benito-Leon J, Louis ED, Villarejo-Galende A, Romero JP, Bermejo-Pareja F. Long sleep duration in elders without dementia increases risk of dementia mortality (NEDICES). Neurology. 2014;83(17): Epub 2014 Sep 24. Agundez JA, Garcia-Martin E, Martinez C, Benito-Leon J, Millan-Pascual J, Calleja P, et al. NQO1 gene rs variant is not associated with risk for multiple sclerosis. BMC neurology. 2014;14:87. Epub 2014 Apr 23. Sierra-Hidalgo F, Llamas S, Gonzalo JF, Sanchez Sanchez C. Ocular dipping in creutzfeldt-jakob disease. Journal of clinical neurology (Seoul, Korea). 2014;10(2): Epub 2014 Apr 23. Anitua E, Pascual C, Antequera D, Bolos M, Padilla S, Orive G, Carro E. Plasma rich in growth factors (PRGF-Endoret) reduces neuropathologic hallmarks and improves cognitive functions in an Alzheimer s disease mouse model. Neurobiology of aging. 2014;35(7): Epub 2014 Jan 17. Gomez C, Vega-Quiroga S, Bermejo-Pareja F, Medrano MJ, Louis ED, Benito-Leon J. Polypharmacy in the Elderly: A Marker of Increased Risk of Mortality in a Population-Based Prospective Study (NEDICES). Gerontology. [Epub ahead of print]. Epub 2014 Dec 6. Leon Ruiz M, Lagares Gomez-Abascal A, Fernandez Alen JA, Benito-Leon J, Garcia-Albea Ristol E. Subarachnoid haemorrhage from a ruptured intracranial mirror-like aneurysm. A case report and literature review. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Aug 22. Agundez JA, Garcia-Martin E, Martinez C, Benito-Leon J, Millan-Pascual J, Diaz-Sanchez M, et al. The GSTP1 gene variant rs1695 is not associated with an increased risk of multiple sclerosis. Cellular & molecular immunology. [Epub ahead of print]. Epub 2014 Dec 22. Olazaran Rodriguez J, Bermejo Pareja F. There is no scientific basis for retiring the MMSE. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 May 15. Romero JP, Benito-Leon J, Mitchell AJ, Trincado R, Bermejo-Pareja F. Under reporting of dementia deaths on death certificates using data from a population-based study (NEDICES). Journal of Alzheimer s disease: JAD. 2014;39(4): Romero JP, Benito-Leon J, Louis ED, Bermejo-Pareja F. Under reporting of dementia deaths on death certificates: a systematic review of population-based cohort studies. Journal of Alzheimer s disease: 29

36 JAD. 2014;41(1): Benito-Leon J, Louis ED, Villarejo-Galende A, Romero JP, Bermejo-Pareja F. Under-reporting of Parkinson s disease on death certificates: A population-based study (NEDICES). Journal of the neurological sciences. 2014;347(1-2): Epub 2014 Oct 2. Benito-Leon J, Labiano-Fontcuberta A, Mitchell AJ, Moreno-Garcia S, Martinez-Martin P. Multiple sclerosis is associated with high trait anger: a case-control study. Journal of the neurological sciences. 2014;340(1-2): Epub 2014 Feb 26. Bolos M, Antequera D, Aldudo J, Kristen H, Bullido MJ, Carro E. Choroid plexus implants rescue Alzheimer s disease-like pathologies by modulating amyloid-β degradation. Cellular and molecular life sciences: CMLS. 2014;71(15): Perez-Gonzalez R, Alvira-Botero MX, Robayo O, Antequera D, Garzon M, Martin-Moreno AM, et al. Leptin gene therapy attenuates neuronal damages evoked by amyloid-β and rescues memory deficits in APP/PS1 mice. Gene therapy. 2014;21(3): Research Projects Code: Title: Análisis de cambios en el patrón de tecleado en una muestra de enfermos de Parkinson Principal Investigator: Félix Bermejo Pareja CIBERNED s groups: Type: International Funding Agency: Massachusetts Institute of Technology Funding: N.D. Duration: Code: Fundación Ramón Areces 2012 Title: Estudio de la capacidad neuroprotectora de las células epiteliales de los plexos coroideos como potencial terapia regenerativa de la enfermedad de Alzheimer Principal Investigator: Eva Carro CIBERNED s collaboration: Yes CIBERNED s groups: G509; G502 Type: National Funding Agency: Fundación Ramón Areces Funding: Duration: Code: MUTUA 2012 Title: Estudio de las propiedades terapéuticas de fármacos basados en Melatonina como tratamiento innovador para enfermedades neurodegenerativas Principal Investigator: Mª Isabel Rodríguez-Franco CIBERNED s groups: Type: International Funding Agency: Fundación Investigación Biomédica de la MUTUA Funding: Duration: Code: Madrid-MIT M+Vision Fellow Title: Motor Response to Dopaminergic Therapy in a Population of De Novo Parkinson s Disease Cases Quantified via Typing Analyses-NeuroQWERTY Principal Investigator: Félix Bermejo-Pareja CIBERNED s groups: Type: International Funding Agency: Madrid-MIT m+vision Consortium Funding: N.D. Duration: Program 1 30

37 2014 Annual Report Félix Bermejo Pareja Group PhD Dissertations Author: Juan Pablo Romero Title: Causas de mortalidad en la cohorte NEDICES. Riesgo inverso entre demencia y cáncer Date: 27th March 2014 Supervisor: Félix Bermejo Pareja Author: Adriana Serna Trujillo Title: Utilidad de una batería neuropsicológica breve para la discriminación de pacientes con demencia y alteración cognitiva leve. Datos de la cohorte NEDICES Date: 1st December 2014 Supervisor: Félix Bermejo Pareja 31

38 Group María Jesús Bullido Gómez Program 1 CONTACT DETAILS oup María Jesús Bullido Gómez Group María Centro Jesús Bullido de Biología Gómez Molecular Group Severo María Jesús Ochoa Bullido (CSIC-UAM) Gómez Group María sús Bullido Gómez Group María Jesús Bullido Gómez Group María Jesús Universidad Bullido Gómez Autónoma Group de Madrid María Jesús Bullido mez Group María Jesús Bullido Gómez Group María Jesús Bullido Gómez Group María Jesús Edificio Bullido CBM Gómez Group Phone: María Jesús Bullido Gómez Group María Jesús Bullido Gómez Group María Jesús Bullido Gómez Group María Jesús Fax: llido Gómez Group María Jesús Bullido Gómez Group María Jesús Bullido Gómez mjbullido@cbm.csic.es Group María Jesús Bullido Gómez oup María Jesús Bullido Gómez Group María Jesús Bullido Gómez Group María Jesús Bullido Gómez Group María LIST OF PERSONNEL María Jesús Bullido Principal Investigator Ana Frank García Head of Department, Researcher Jesús Aldudo Soto Researcher María Recuero Vicente Researcher Isabel Sastre Merlín Technician Henrike Kristen PhD student Manuel Manchón Romero Master s student Julia Terreros Roncal Student 32

39 2014 Annual Report María Jesús Bullido Gómez Group Summary To identify genes and mechanisms involved in the neurodegeneration associated with Alzheimer s disease (AD), we developed cellular models of oxidative stress (OS) and HSV 1 infection,that show changes characteristic of AD as those related with autophagy, tau protein phosphorylation and β-amyloid precursor protein (APP) proteolysis. During this period, we have demonstrated that the regulation of APP metabolism by OS involves the two major cellular proteolytic machineries: the ubiquitin/proteasome and the autophagy-lysosome. We have also shown that OS exacerbates the effects of HSV 1 via complex interactions that include the inhibition of viral replication. Both HSV 1 and OS block the fusion of autophagosomes with lysosomes, and can in this way cause the observed alterations. Using gene expression studies we have identified a set of genes activated by OS in infected cells and in a cell model of familial AD. Its pathway enrichment analysis suggests that the interaction of OS with HSV 1 or with mutations causing familial AD provokes alterations of the lysosomal function. These data support the findings mentioned above and those of other authors, highlighting the role of lysosomes in different forms of neurodegeneration. Several data obtained in patients by genetic association studies of our group and of different collaborative projects in which we participate, as the IGAP consortium, also support this implication. Remarkably, a recent IGAP pathway enrichment analysis of genomic data from about 80,000 participants drew endocytosis and protein ubiquitination, processes that converge in the lysosome, as prime therapeutic targets for AD. As for studies in patients, we continue to participate in genetic association projects in collaboration with CIBERNED groups, as well as in the EADI e IGAP consortia, which are showing new factors and functions potentially relevant in the pathogenesis of AD. We have also continued genetic association studies in the Spanish Group for Genetic Study of Dementia (DEGESCO), which have led so far to an article published in 2014, one under review and two in preparation. We have also continued the longitudinal study of patients with mild cognitive impairment, in collaboration with the Foundations CIEN and Reina Sofía. The correlation analysis of the neuropsychological, neuroimaging, biochemical, genetic and clinical variables, still in development, has led in the last year to the first interesting findings, like the description of a biomarker panel that may facilitate the specific detection of early-stage AD through the analysis of plasma samples. Keywords Alzheimer, Herpes virus, Oxidative stress, APP metabolism, Functional genomics, Genetic association, Biomarkers Publications Sanchez-Benavides G, Pena-Casanova J, Casals-Coll M, Gramunt N, Molinuevo JL, Gomez-Anson B, et al. Cognitive and neuroimaging profiles in mild cognitive impairment and Alzheimer's disease: data from the Spanish Multicenter Normative Studies (NEURONORMA Project). Journal of Alzheimer's disease: JAD. 2014;41(3):

40 International Genomics of Alzheimer's Disease Consortium (IGAP), International Genomics of Alzheimer's Disease Consortium IGAP. Convergent genetic and expression data implicate immunity in Alzheimer's disease. Alzheimer's & dementia: the journal of the Alzheimer's Association. [Epub ahead of print]. Epub 2014 Dec 20. Manero RM, Casals-Coll M, Sanchez-Benavides G, Rodriguez-de los Reyes ON, Aguilar M, Badenes D, et al. Diagnostic validity of the Alzheimer's disease functional assessment and change scale in mild cognitive impairment and mild to moderate Alzheimer's disease. Dementia and geriatric cognitive disorders. 2014;37(5-6): Epub 2014 Feb 18. Escott-Price V, Bellenguez C, Wang LS, Choi SH, Harold D, Jones L, et al. Gene-wide analysis detects two new susceptibility genes for Alzheimer's disease. PloS one. 2014;9(6):e Epub 2014 Jun 12. Bolos M, Antequera D, Aldudo J, Kristen H, Bullido MJ, Carro E. Choroid plexus implants rescue Alzheimer's disease-like pathologies by modulating amyloid-β degradation. Cellular and molecular life sciences: CMLS. 2014;71(15): Epub 2013 Dec 17. Casals-Coll M, Sanchez-Benavides G, Meza-Cavazos S, Manero RM, Aguilar M, Badenes D, et al. Spanish multicenter normative studies (NEURONORMA project): normative data and equivalence of four BNT short-form versions. Archives of clinical neuropsychology: the official journal of the National Academy of Neuropsychologists. 2014;29(1): Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodriguez-Rodriguez E, Lopez de Munain A, et al. Assessing the role of the TREM2 p.r47h variant as a risk factor for Alzheimer's disease and frontotemporal dementia. Neurobiology of aging. 2014;35(2):444.e1-4. Program 1 34

41 Group Antonio Camins Espuny Program 1 CONTACT DETAILS Group Antonio Camins Espuny Group Antonio Camins Espuny Group Antonio Camins Facultad Espuny de Farmacia Group Antonio Camin Espuny Group Antonio Camins Espuny Group Antonio Departamento Camins Espuny de Farmacología Group Antonio y Química Camins Terapéutica Espuny Group Antoni Universidad de Barcelona Camins Espuny Group Antonio Camins Espuny Group Antonio Camins Espuny Group Av. Antonio Joan XXIII, Camins s/n Espuny Group Antonio Camins Espuny Group Antonio Camins Espuny Group Antonio Camins Espuny Barcelona Group (Spain) Antonio Camins Espun Phone: Group Antonio Camins Espuny Group Antonio Camins Espuny Group Antonio Camins camins@ub.edu Espuny Group Antonio Camin Espuny Group Antonio Camins Espuny Group Antonio Camins Espuny Group Antonio Camins Espuny Group Antoni LIST OF PERSONNEL Antoni Camins Espuny Principal Investigator, Full Professor Merce Pallàs Lliberia Full Professor Jordi Vilaplana Associate Professor Carme Pelegrí Associate Professor Francesc Sureda Associate Professor Carme Auladell Associate Professor Jaume Folch Associate Professor Anna Maria Canudas Teixidó Associate Professor Andrés Jiménez Associate Professor Ester Verdaguer Cardona Lecturer Mirem Ettecheto Research Fellow Gemma Manich Research Fellow Itsaso Cabezón Rodríguez Research Fellow Ivan Patraca Research Fellow Nacho Pedrós Research Fellow Victor Rimbau Research Fellow Christian Grañe Farre Research Fellow Sonia Abad Research Fellow 35

42 Summary The aims of our research is the study in experimental models of Alzheimer s Disease (APPSwe/PS1dE9) and ageing (SMP8) the molecular pathways involved in neurodegeneration and again. We study potential pathways such as mitogen activated protein kinases (JNK1, JNK2, JNK3), cdk5, etc. involved in cell death and tau phosphorylation. In addition our interest is the development of therapeutic strategies for Alzheimer s disease treatment. Thus, in recent years we have demonstrated in the APPSwe/PS1dE9 mice model of Alzheimer s a relationship between alterations in the metabolism specifically, type 2 diabetes mellitus and an advancement and contributes to development and progression of the Alzheimer s disease. Thus treatment of APPSwe/PS1dE9 mice with a high fat diet favors disease worsening and memory loss. Currently, we have demonstrated changes in transcripts and proteins involved in the insulin pathway in the hippocampus of the APPSwe/PS1dE9 mice a familial model of Alzheimer s disease. Moreover, we have detected disturbances in intracellular signalling pathways downstream of prolactin and leptin receptors, JAK/STAT signalling, as well as abnormalities in cholesterol metabolism in the hippocampi of APPSwe/PS1dE9 mice between the 3 and 6 months of age. Alzheimer s disease is also associated with the decrease in insulin levels in the central nervous system and alterations of insulin receptors and downstream pathway in the brain. Defects in insulin signaling/igf-1 affect PI3K/Akt causing neurodegeneration process. Besides the alteration of glucose transporters at the brain or decrease its expression in the brain of Alzheimer s patients may cause a lower glucose metabolism in the brain and cause lower reduced mitochondrial metabolism and ATP production. Resveratrol is a natural product that is of great interest as anti-aging and neuroprotective therapy based on is properties as an activator of sirtuin 1. The sirtuins are a family of highly conserved proteins in both eukaryotic and prokaryotic organisms that have a very important role in the aging process and in the regulation of fundamental physiological processes. The interest of our group is focused on studying the role of sirtuin 1 (SIRT1) and other sirtuins as SIRT3 in the process of aging and Alzheimer s disease in APPswe/PS1dE9 and accelerated aging mice (SAMP8). In addition we would like to evaluate the role of SIRT1 in cultured neurons to demonstrate the involvement of this enzyme in the process of neuroprotection preventing the neuronal process of apoptosis through regulating p53, ATM and other proteins involved in the process of neuronal death. Another interesting point is the study of the role of exercise in the prevention of aging and neuronal death in mice SAMP8 and SAMR1. We are studding the possible biochemical pathways involved in the beneficial effect of exercise in rodents. Thus, a study has been performed in these mice evaluating the expression of genes modulated by the exercise that could have a potential beneficial effect, as anti-aging and also exerts a possibly neuroprotective role. Characterization of a neo-epitope present in brain granule structures related to aging in mice with accelerated senescence SAMP8. These structures appear more amount and frequency in the hippocampus of aged animals and especially in animals strain SAMP8. These granules contain remains of membranes and cell organelles and signs of degeneration as autophagosomes and large vacuoles. The study of the neo-epitope of the granules can provide information about the formation of these structures during the aging and neurodegenerative processes. Furthermore, we are also working on developing new synthetic molecules for the potential treatment of Alzheimer s disease. Program 1 36

43 2014 Annual Report Antonio Camins Espuny Group Keywords Alzheimer, APPswe/PS1dE9, SAMP8, Resveratrol, Aging, Diabetes Publications Abad S, Junyent F, Auladell C, Pubill D, Pallàs M, Camarasa J, et al. 3,4-Methylenedioxymethamphetamine enhances kainic acid convulsive susceptibility. Progress in neuro-psychopharmacology & biological psychiatry. 2014;54: Epub 2014 Jun 28. Manich G, Cabezon I, Camins A, Pallàs M, Liberski PP, Vilaplana J, et al. Clustered granules present in the hippocampus of aged mice result from a degenerative process affecting astrocytes and their surrounding neuropil. Age (Dordrecht, Netherlands). 2014;36(4):9690. Epub 2014 Jul 30. Bayod S, Felice P, Andres P, Rosa P, Camins A, Pallàs M, et al. Downregulation of canonical Wnt signaling in hippocampus of SAMP8 mice. Neurobiology of aging. 2015;36(2): Epub 2014 Sep 28. Pedros I, Petrov D, Allgaier M, Sureda F, Barroso E, Beas-Zarate C, et al. Early alterations in energy metabolism in the hippocampus of APPswe/PS1dE9 mouse model of Alzheimer's disease. Biochimica et biophysica acta. 2014;1842(9): Epub 2014 Jun 2. Lalanza JF, Caimari A, del Bas JM, Torregrosa D, Cigarroa I, Pallàs M, et al. Effects of a post-weaning cafeteria diet in young rats: metabolic syndrome, reduced activity and low anxiety-like behaviour. PloS one. 2014;9(1):e Epub 2014 Jan 15. Cosin-Tomas M, Alvarez-Lopez MJ, Sanchez-Roige S, Lalanza JF, Bayod S, Sanfeliu C, et al. Epigenetic alterations in hippocampus of SAMP8 senescent mice and modulation by voluntary physical exercise. Frontiers in aging neuroscience. 2014;6:51. Epub 2014 Mar 20. Bayod S, Del Valle J, Pelegri C, Vilaplana J, Canudas AM, Camins A, et al. Macroautophagic process was differentially modulated by long-term moderate exercise in rat brain and peripheral tissues. Journal of physiology and pharmacology: an official journal of the Polish Physiological Society. 2014;65(2): Petrov D, Pedros I, de Lemos ML, Pallàs M, Canudas AM, Lazarowski A, et al. Mavoglurant as a treatment for Parkinson's disease. Expert opinion on investigational drugs. 2014;23(8): Epub 2014 Jun 24. Ettcheto M, Junyent F, de Lemos L, Pallas M, Folch J, Beas-Zarate C, et al. Mice Lacking Functional Fas Death Receptors Are Protected from Kainic Acid-Induced Apoptosis in the Hippocampus. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Aug 15. Porquet D, Grinan-Ferre C, Ferrer I, Camins A, Sanfeliu C, Del Valle J, et al. Neuroprotective role of trans-resveratrol in a murine model of familial Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;42(4): Valverde E, Sureda FX, Vazquez S. Novel benzopolycyclic amines with NMDA receptor antagonist activity. Bioorganic & medicinal chemistry. 2014;22(9): Epub 2014 Mar 24. Ortuno-Sahagun D, Pallàs M, Rojas-Mayorquin AE. Oxidative stress in aging: advances in proteomic approaches. Oxidative medicine and cellular longevity. 2014;2014: Epub 2014 Feb 13. Rivera-Cervantes MC, Castaneda-Arellano R, Castro-Torres RD, Gudino-Cabrera GY, Velasco AI, Camins A, et al. P38 MAPK Inhibition Protects Against Glutamate Neurotoxicity and Modifies NMDA and AMPA Receptor Subunit Expression. Journal of molecular neuroscience: MN. 2015;55(3): Epub 2014 Aug 30. Alvarez-Lopez MJ, Molina-Martinez P, Castro-Freire M, Cosin-Tomas M, Cristòfol R, Parrizas M, et al. Rcor2 underexpression in senescent mice: a target for inflammaging? Journal of neuroinflammation. 2014;11:126. Epub 2014 Jul

44 Pallàs M, Ortuno-Sahagun D, Benito-Andres P, Ponce-Regalado MD, Rojas-Mayorquin AE. Resveratrol in epilepsy: preventive or treatment opportunities? Frontiers in bioscience: a journal and virtual library. 2014;19: Epub 2014 Jun 1. Pereira NA, Sureda FX, Esplugas R, Perez M, Amat M, Santos MM. Tryptophanol-derived oxazolopiperidone lactams: identification of a hit compound as NMDA receptor antagonist. Bioorganic & medicinal chemistry letters. 2014;24(15): Epub 2014 Jun 11. Bayod S, Guzman-Brambila C, Sanchez-Roige S, Lalanza JF, Kaliman P, Ortuno-Sahagun D, et al. Voluntary Exercise Promotes Beneficial Anti-aging Mechanisms in SAMP8 Female Brain. Journal of molecular neuroscience: MN. 2014;55(2): Epub 2014 Jul 16. Mancuso R, Del Valle J, Morell M, Pallas M, Osta R, Navarro X. Lack of synergistic effect of resveratrol and sigma-1 receptor agonist (PRE-084) in SOD1 G93A ALS mice: overlapping effects or limited therapeutic opportunity? Orphanet journal of rare diseases. 2014;9:78. Epub 2014 May 21. Mancuso R, del Valle J, Modol L, Martinez A, Granado-Serrano AB, Ramirez-Nunez O, et al. Resveratrol improves motoneuron function and extends survival in SOD1(G93A) ALS mice. Neurotherapeutics: the journal of the American Society for Experimental NeuroTherapeutics. 2014;11(2): Manich G, del Valle J, Cabezon I, Camins A, Pallàs M, Pelegri C, et al. Presence of a neo-epitope and absence of amyloid beta and tau protein in degenerative hippocampal granules of aged mice. Age (Dordrecht, Netherlands). 2014;36(1): Vilar A, de Lemos L, Patraca I, Martinez N, Folch J, Junyent F, et al. Melatonin suppresses nitric oxide production in glial cultures by pro-inflammatory cytokines through p38 MAPK inhibition. Free radical research. 2014;48(2): Castro-Torres RD, Chaparro-Huerta V, Flores-Soto ME, Banuelos-Pineda J, Camins A, Orozco-Suarez SA, et al. A single dose of pirfenidone attenuates neuronal loss and reduces lipid peroxidation after kainic acid-induced excitotoxicity in the pubescent rat hippocampus. Journal of molecular neuroscience: MN. 2014;52(2): Bayod S, Menella I, Sanchez-Roige S, Lalanza JF, Escorihuela RM, Camins A, et al. Wnt pathway regulation by long-term moderate exercise in rat hippocampus. Brain research. 2014;1543: Abad S, Fole A, del Olmo N, Pubill D, Pallàs M, Junyent F, et al. MDMA enhances hippocampaldependent learning and memory under restrictive conditions, and modifies hippocampal spine density. Psychopharmacologia. 2014;231(5): Ortuno-Sahagun D, Gonzalez RM, Verdaguer E, Huerta VC, Torres-Mendoza BM, Lemus L, et al. Glutamate excitotoxicity activates the MAPK/ERK signaling pathway and induces the survival of rat hippocampal neurons in vivo. Journal of molecular neuroscience: MN. 2014;52(3): Redondo-Castro E, Romero R, Torres-Espin A, Utrera J, Duque D, Junyent F, et al. Dithiocarb (N,Ndiethyldithiocarbamate, DEDTC) decreases levels of biogenic monoamines in the adult mouse brain. Neuropathology and applied neurobiology. 2014;40(6): Research Projects Code: BFU P Title: Caracterización de un neo-epítopo cerebral relacionado con el envejecimiento y estudio de la presencia de anticuerpos naturales anti-neoepítopo Principal Investigator: Carme Pelegrí Gavalda CIBERNED s groups: Type: National Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: T12116S Title: Determination of antagonist activity of new compounds at the nmda receptor Principal Investigator: Francesc X. Sureda CIBERNED s groups: Type: International Program 1 38

45 2014 Annual Report Antonio Camins Espuny Group Funding Agency: Faculdade de Farmacia da Universidade de Lisboa Funding: Duration: Code: SAF Title: Implicación de la isoforma Jnk3 en la enfermedad de Alzheimer. Evaluación de su potencial como diana terapéutica Principal Investigator: Antonio Camins Espuny CIBERNED s groups: Type: National Funding Agency: Ministerio de Ciencia e Innovación Funding: Duration: Code: CIBERNED 2013/05 Title: Identificación y caracterización molecular de subpoblaciones de receptores cannabinoides en poliglutaminopatías Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes CIBERNED s groups: G201; G303; G304 Type: International Funding Agency: CIBERNED Funding: Duration: Code: FC Title: Cofinanciamiento de Becas IRTA-URVConvenio IRTA-URV Banco de Santander Principal Investigator: Francesc X. Sureda CIBERNED s groups: Type: International Funding Agency: Institut de Recerca i Tecnologies Agroalimentàries (IRTA) Funding: Duration: 2014 PhD Dissertations Author: Gemma Manich Title: Caracterització de les estructures granulars patològiques presents a l'hipocamp dels ratolins amb senescència accelerada SAMP8 Date: 25th july 2014 Supervisor: Carmen Pelegrí Author: David Porquet Title: Efecto del resveratrol en modelos murinos de envejecimiento y enfermedad de Alzheimer Date: 1st july2014 Supervisor: Mercedes Pallàs Lliberia 39

46 Group José Luis Cantero Lorente Program 1 CONTACT DETAILS oup José Luis Cantero Lorente Group José Luis Cantero Lorente Laboratorio Group José de Neurociencia Luis Cantero Lorente Funcional Group José Luis ntero Lorente Group José Luis Cantero Lorente Group Servicios José Centralizados Luis Cantero de Lorente Investigación Group (Edificio José Luis 21) Cantero Lorente Universidad Pablo de Olavide oup José Luis Cantero Lorente Group José Luis Cantero Lorente Group José Luis Ctra. Cantero de Utrera, Lorente Km. Group 1 José Luis ntero Lorente Group José Luis Cantero Lorente Group José Luis Cantero Lorente Group Sevilla José (Spain) Luis Cantero Lorente Phone: oup José Luis Cantero Lorente Group José Luis Cantero Lorente Group José Luis Fax: Cantero Lorente Group José Luis jlcanlor@upo.es ntero Lorente Group José Luis Cantero Lorente Group José Luis Cantero Lorente Group José Luis Cantero Lorente LIST OF PERSONNEL José Luis Cantero Lorente Principal Investigator Mercedes Atienza Ruiz Researcher Maité Crespo García Postdoctoral Fellow Gabriel González Escamilla PhD student Mayely Sánchez Espinosa PhD student Sofia Rodríguez Peñuela Technician David García Solís Researcher Eulogio Gil Néciga Researcher 40

47 2014 Annual Report Group José Luis Cantero Lorente Summary In 2014, our group has examined anatomo-functional correlates of sleep deficits in elderly subjects with mild cognitive impairment (MCI). This work has established that slow-wave sleep fragmentation shown by MCI subjects is associated with increased levels of plasma amyloid-beta 42, whereas reduced REM sleep is related to thinning of the precuneus and posterior region of the cingulate cortex (Sanchez- Espinosa et al., 2014). Together, these results suggest that the relationship between amyloid-beta levels and impaired sleep in elderly subjects with MCI is similar to the one recently reported in animal models of Alzheimer s disease (AD). In the view of the close link between sleep, memory and AD, we have further investigated in young subjects whether encoding and consolidation processes are significantly affected by sleep deprivation depending on the degree of hippocampal dependence. Results have shown that acute sleep restriction impairs encoding more than consolidation of hippocampal-dependent memories (Alberca-Reina et al., 2014). On the other hand, we have compared neuropsychiatric symptomatology between dementia with Lewy bodies and AD patients, since these manifestations are one of the main features for the differential diagnosis of these two neurodegenerative diseases. We found that hallucinations and delusions are not only more frequent in dementia with Lewy bodies than in AD patients but also have distinct qualitative characteristics and patterns of progression that can be helpful to enhance differential diagnosis (Suarez- Gonzalez et al., 2014). Finally, we have explored how aging modifies the relationship between structure and function in cortical networks, based on patterns of cortical thickness and glucose intake. Our results showed that the relationship between structural and functional cortical connectivity patterns in aging is mainly constrained by functional connectivity at local level, likely due to aging-related deficits in functional integration. Once aging processes weaken these connections, coupling between structural and cortical networks is unbalanced and topological organization of the neocortex becomes more inefficient (Romero-Garcia et al., 2014). Keywords Cerebral aging, Neuroimaging and cerebral electrophysiology markers of prodromal and asymptomatic stages of Alzheimer s disease, Anatomical and functional connectivity of human neocortex, Computational neuroanatomy techniques applied to incipient neurodegeneration Publications Crespo-Garcia M, Hartmann T. Does illusory flickering result from rhythmic sampling of visual stimuli? The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(2): Garcia-Gomez FJ, Marin-Oyaga VA, Moniche-Alvarez F, Vazquez-Albertino RJ, Garcia-Solis D. In Process Citation. Revista de neurologia. 2014;58(10): Huertas-Fernandez I, Garcia-Gomez FJ, Garcia-Solis D, Benitez-Rivero S, Marin-Oyaga VA, Jesus S, et al. Machine learning models for the differential diagnosis of vascular parkinsonism and Parkinson's disease using (123)IFP-CIT SPECT. European journal of nuclear medicine and molecular imaging. 2015;42(1): Epub 2014 Aug 14. Romero-Garcia R, Atienza M, Cantero JL. Predictors of coupling between structural and functional cortical networks in normal aging. Human brain mapping. 2014;35(6): Epub 2013 Sep

48 Suarez-Gonzalez A, Heredia CG, Savage SA, Gil-Neciga E, Garcia-Casares N, Franco-Macias E, et al. Restoration of conceptual knowledge in a case of semantic dementia. Neurocase. 2015;21(3):1-13. Epub 2014 Mar 5. Alberca-Reina E, Cantero JL, Atienza M. Semantic congruence reverses effects of sleep restriction on associative encoding. Neurobiology of learning and memory. 2014;110: Epub 2014 Jan 24. Sanchez-Espinosa MP, Atienza M, Cantero JL. Sleep deficits in mild cognitive impairment are related to increased levels of plasma amyloid-β and cortical thinning. NeuroImage. 2014;98: Epub 2014 May 16. Suarez-Gonzalez A, Serrano-Pozo A, Arroyo-Anllo EM, Franco-Macias E, Polo J, Garcia-Solis D, et al. Utility of neuropsychiatric tools in the differential diagnosis of dementia with Lewy bodies and Alzheimer's disease: quantitative and qualitative findings. International psychogeriatrics / IPA. 2014;26(3): Epub 2013 Nov 28. Martinez Castillo R, Fernandez Lopez R, Acevedo Banez I, Alvarez Perez RM, Garcia Solis D, Vazquez Albertino R, et al. Utility of single photon emission computed tomography-computed tomography in selective sentinel lymph node biopsy in patients with melanoma. Revista espanola de medicina nuclear e imagen molecular. 2014;33(3): Epub 2013 Oct 2. Arbizu J, Luquin MR, Abella J, de la Fuente-Fernandez R, Fernandez-Torron R, Garcia-Solis D, et al. Functional neuroimaging in the diagnosis of patients with Parkinsonism: Update and recommendations for clinical use. Revista espanola de medicina nuclear e imagen molecular. 2014;33(4): Epub 2014 Apr 14. Research Projects Code: P12-CTS 2327 Title: Caracterización de las fases presintomáticas y preclínicas de la enfermedad de Alzheimer mediante marcadores biológicos y de neuroimagen Principal Investigator: Jose Luis Cantero Lorente CIBERNED s groups: Type: International Funding Agency: Consejería de Economía, Innovación y Ciencia, Junta de Andalucía Funding: 225,419 Duration: Code: SAF Title: Combining beta-amyloid biomarkers with anatomo-functional neuroimaging to predict cognitive decline in presymptomatic stages of Alzheimer s disease Principal Investigator: Jose Luis Cantero Lorente CIBERNED s groups: Type: National Funding Agency: Ministerio de Ciencia e Innovación, Plan Nacional I+D+I, Program Nacional de Biomedicina Funding: Duration: Code: 2013 Title: Deterioro de la conectividad anatómica asociado a las fases presintomáticas de la enfermedad de Alzheimer: De la microestructura de la sustancia blanca cerebral a los cambios en los haces de fibras nerviosas Principal Investigator: Jose Luis Cantero Lorente CIBERNED s groups: Type: Private Funding Agency: Instituto de Especialidades Neurológicas (IENSA) Funding: Duration: Code: PSI Title: Papel de las oscilaciones cerebrales del sueño no REM en la codificación y consolidación de memorias asociativas Principal Investigator: Mercedes Atienza Ruiz CIBERNED s groups: Type: National Funding Agency: MICINN, Plan Nacional I+D+i Funding: Duration: Program 1 42

49 2014 Annual Report Group José Luis Cantero Lorente PhD Dissertations Author: Eva Hita-Yáñez Title: Caracterización de las alteraciones del sueño en personas mayores con deterioro cognitivo leve Date: 17th January 2014 Supervisor: José Luis Cantero Lorente Author: Kelly Cristina Atalaia da Silva Title: Correlatos neuroanatómicos del déficit de memoria episódica en personas mayores con deterioro cognitivo leve Date: 31st January 2014 Supervisor: José Luis Cantero Lorente Author: Rafael Romero-García Title: Organización topológica de la corteza cerebral en el envejecimiento normal y en la enfermedad de Alzheimer Date: 21st November 2014 Supervisor: José Luis Cantero Lorente 43

50 Group Joan Xavier Comella Carnicé Program 1 CONTACT DETAILS oup Joan Xavier Comella Carnicé Group Joan Xavier Comella Carnicé VHIR Group Vall Hebron Joan Xavier Institut Comella de Recerca Carnicé Group Joan vier Comella Carnicé Group Joan Xavier Comella Carnicé Group Joan Passeig Xavier Comella Vall d Hebron Carnicé Group Joan Xavier mella Carnicé Group Joan Xavier Comella Carnicé Group Joan Xavier Comella Carnicé Barcelona Group (Spain) Joan Xavier Comella rnicé Group Joan Xavier Comella Carnicé Group Joan Xavier Comella Carnicé Phone: Group +34 Joan Xavier 807Comella Carnicé oup Joan Xavier Comella Carnicé Group Joan Xavier Comella Carnicé Group Joan Xavier Comella Carnicé Group joan.comella@vhir.org an Xavier Comella Carnicé Group Joan Xavier Comella Carnicé Group Joan Xavier Comella Carnicé Group Joan Xavier LIST OF PERSONNEL Joan Xavier Comella Carnicé Principal Investigator Víctor Yuste Mateos PhD Researcher Joaquín López Soriano PhD Researcher Bruna Barneda Zahonero PhD Researcher Jorge Urresti Ibáñez PhD Researcher Laura Planells Ferrer PhD Researcher María Sánchez Osuna PhD Researcher Koen Galenkamp PhD student Isabel Calleja Yagüe PhD student Elena Coccia Master student 44

51 2014 Annual Report Joan Xavier Comella Carnicé Group Summary Our main interest is to characterize the mechanisms controlling neuronal death induced by a group of receptors collectively known as death receptors, and also the relevance of some of their intracellular antagonists. Among those we are most interested in the antagonists expressed in nervous system, such as FAIM-L and Lifeguard. Our hypothesis is that Alzheimer s disease (AD) and other neurodegenerative diseases have a neuroinflammatory component. In early stages, glial activation plays a neuroprotective role, executed by cytokines such as TNF, which induces survival pathways. In advanced stages, chronic neuroinflammation activates death receptors with a loss of the protective role of their functional antagonists (such as FAIM-L) and thus promoting neuronal death. Thus, our main objective is the molecular characterization of these death receptor antagonists, their role in neuronal differentiation and physiology, and their involvement in different pathologies, particularly neurodegenerative diseases. In addition, we are characterizing molecular partners of these molecules that may explain their mechanism of action. In this sense, during 2013 we described in detail the interaction between FAIM-L and XIAP, in an article published at the Journal of Neuroscience. At present, we are characterizing the interaction of FAIM-L with another protein, Siva-1, which in turn has been implicated in apoptotic processes and which interacts with XIAP. We are studying its likely implications in neuron death, but also in neuron plasticity processes, where non-apoptotic activities of caspase-3 are involved (being its activity inhibited by XIAP). Another of our main research lines, in collaboration with different CIBERNED groups, aims to characterize the role of FAIM-L in both animal models (APP/PS1) and patients of Alzheimer s disease. We have characterized that FAIM-L levels are downregulated in AD patients and in the APP/PS1 model, and that beta-amyloid reduces FAIM-L levels in neurons in vitro. Additionally, TNF is able to protect from the betaamyloid-induced cell death in vitro, only if neurons have enough levels of FAIM-L. Thus, FAIM-L levels can determine the cell fate in a neuroinflammatory process. These results have been recently published at Cell Death & Disease. The knowledge of the molecular mechanisms of action of these death receptor antagonists and how they induce survival signaling pathways could be highly relevant to a better understanding of the etiology and also to open new strategies to improve the treatment of some neurodegenerative illnesses, such as AD or Parkinson s disease, as evidenced with our results on FAIM-L. Our main research lines at present are: To study FAIM-L function in vitro and in vivo models of AD, and its relation with TNF (its duality as a pro-apoptotic molecule and at the same time as a survival promoter) and its signaling pathways. To characterize FAIM-L functional partners and their implications in neuron physiology (neuronal plasticity) and different neurodegenerative diseases (neuronal apoptosis, synaptic degeneration). To study Faim promoter and the functional differences of FAIM isoforms. Keywords Alzheimer, Neuroinflammation, Death receptors, FAIM, Lifeguard, TNF, Fas 45

52 Publications Moreno M, Negrotto L, Rio J, Moubarak R, Martin I, Bustamante MF, et al. Activation-induced cell death in T lymphocytes from multiple sclerosis patients. Journal of neuroimmunology. 2014;272(1-2):51-5. Epub 2014 Apr 16. Carriba P, Comella JX. Amyloid Beta, TNFα and FAIM-L; Approaching New Therapeutic Strategies for AD. Frontiers in neurology. 2014;5:276. Epub 2014 Dec 18. Cornago M, Garcia-Alberich C, Blasco-Angulo N, Vall-Llaura N, Nager M, Herreros J, et al. Histone deacetylase inhibitors promote glioma cell death by G2 checkpoint abrogation leading to mitotic catastrophe. Cell death & disease. 2014;5:e1435. Epub 2014 Oct 2. Planells-Ferrer L, Urresti J, Soriano A, Reix S, Murphy DM, Ferreres JC, et al. MYCN repression of Lifeguard/FAIM2 enhances neuroblastoma aggressiveness. Cell death & disease. 2014;5:e1401. Epub 2014 Sep 4. Research Projects Code: SAF R Title: Neuroinflamación, TNF y antagonistas de los receptores de muerte (FAIM-L): relevancia en neurodegeneración Principal Investigator: Joan Xavier Comella Carnicé CIBERNED s groups: Type: National Funding Agency: MINECO Funding: Duration: Code: 2014 SGR 1609 Title: Apoptosi i Neurodegeneracio (GRC) Principal Investigator: Joan Comella Carnicé CIBERNED s collaboration: Yes CIBERNED s groups: G413; G109 Type: International Funding Agency: AGAUR Funding: Duration: Code: PI2013/01 Title: Propiedades emergentes de la relación neurona-glía que subyacen a neurodegeneración y demencia en la enfermedad de Alzheimer Principal Investigator: Ignacio Torres-Alemán CIBERNED s collaboration: Yes CIBERNED s groups: G413; G204; G409; G108; G411; G415 Type: National Funding Agency: CIBERNED Funding: Duration: Code: PIE13/00027 Title: Preventing cardiovascular ischemic events and arresting their consequences in type 2 diabetic population: a multidisciplinary clinical and experimental approach Principal Investigator: David Garcia-Dorado CIBERNED s groups: Type: National Funding Agency: MINECI/ISCIII Funding: Duration: Program 1 46

53 2014 Annual Report Joan Xavier Comella Carnicé Group PhD Dissertations Author: Laura Planells Ferrer Title: Implication of Death Receptor Antagonists in Neuroblastoma: Role of Lifeguard Date: 7th November 2014 Supervisor: Joan Xavier Comella Carnicé Author: Jorge Urresti Ibáñez Title: Characterization of the death receptor antagonist in the nervous system, Lifeguard Date: 28th November 2014 Supervisor: Joan Xavier Comella Carnicé 47

54 Group Javier de Felipe Oroquieta Program 1 CONTACT DETAILS oup Javier de Felipe Oroquieta Group Javier de Felipe Oroquieta Laboratorio Group Cajal Javier de Circuitos de Felipe Corticales, Oroquieta CTB Group Javier de lipe Oroquieta Group Javier de Felipe Oroquieta Group Javier de Felipe Universidad Oroquieta Politécnica Group Javier de Madrid de Felipe Oroquieta Campus de Montegancedo oup Javier de Felipe Oroquieta Group Javier de Felipe Oroquieta Group Javier de Felipe Pozuelo Oroquieta de Alarcón Group Javier de lipe Oroquieta Group Javier de Felipe Oroquieta Group Javier de Felipe Oroquieta Group Madrid Javier (Spain) de Felipe Oroquieta Phone: oup Javier de Felipe Oroquieta Group Javier de Felipe Oroquieta Group Javier de defelipe@cajal.csic.es Felipe Oroquieta Group Javier de lipe Oroquieta Group Javier de Felipe Oroquieta Group Javier de Felipe Oroquieta Group Javier de Felipe Oroquieta LIST OF PERSONNEL Javier de Felipe Oroquieta Principal Investigator, Research Professor Lidia Alonso-Nanclares Postdoctoral researcher Ruth Benavides-Piccione Postdoctoral researcher Rodrigo Rodríguez Sánchez CSIC researcher Isabel Fernaud Espinosa Postdoctoral researcher Asta Kastanauskaite Postdoctoral researcher Ángel Merchán-Pérez Postdoctoral researcher Alberto Muñoz Postdoctoral researcher Gonzalo León Espinosa Postdoctoral researcher Lorena Valdés Lora Technician Ana Isabel García Ramírez Technician Débora Cano Laiseca Technician Mari Carmen Álvarez Pérez Technician Silvia Tapia González Technician Mirian Marín Horcajada Technician Alejandro Antón Fernández PhD student Liulia Diana Furcila PhD student Andrea Santuy Muñoz PhD student Guillermo Aparicio Torres Research Fellow Marta Álvarez Almazán Research Fellow Sandra Sáez Raspeño Research Fellow Miguel Miguens Vázquez Visiting scientist Juncal González Visiting scientist Pilar Flores Romero Research and Project Manager Montserrat Fernández Bouzo Research and Project Manager 48

55 2014 Annual Report Javier de Felipe Oroquieta Group Summary The two main lines of research that we have developed are: Neurochemical and microanatomical analysis of the cerebral cortex in different mouse models for Alzheimer s disease, with the idea of obtaining data about the substrate and temporal course of the microanatomical alterations that occur in Alzheimer s disease. Neurochemical and microanatomical analysis of the cerebral cortex of patients with Alzheimer s disease. In particular, we are performing a quantitative and qualitative analysis of the synaptic circuits and neurochemical characteristics of the cerebral cortex of these patients. The aim of this objective is to try to know in detail the alterations of the neuronal circuits in relation with the senile plaques and the presence of neurofibrillary tangles. Keywords Cerebral cortex, Microorganization, Neuronal circuits, Electron microscopy, Alzheimer Publications Alonso-Nanclares L, DeFelipe J. Alterations of the microvascular network in the sclerotic hippocampus of patients with temporal lobe epilepsy. Epilepsy & behavior: E&B. 2014;38C: Epub 2014 Jan 6. Jimenez-Mateos EM, Engel T, Merino-Serrais P, Fernaud-Espinosa I, Rodriguez-Alvarez N, Reynolds J, et al. Antagomirs targeting microrna-134 increase hippocampal pyramidal neuron spine volume in vivo and protect against pilocarpine-induced status epilepticus. Brain structure & function. [Epub ahead of print]. Epub 2014 May 30. Lopez-Cruz PL, Larranaga P, DeFelipe J, Bielza C. Bayesian network modeling of the consensus between experts: An application to neuron classification. International Journal of Approximate Reasoning. 2014;55(1):3-22. Bielza C, Benavides-Piccione R, Lopez-Cruz P, Larranaga P, DeFelipe J. Branching angles of pyramidal cell dendrites follow common geometrical design principles in different cortical areas. Scientific reports. 2014;4:5909. Epub 2014 Aug 1. De Felipe J. El Jardin de la Neurologia Toharia P, Morales J, de Juan O, Fernaud I, Rodriguez A, DeFelipe J. Musical representation of dendritic spine distribution: a new exploratory tool. Neuroinformatics. 2014;12(2): Morales J, Benavides-Piccione R, Dar M, Fernaud I, Rodriguez A, Anton-Sanchez L, et al. Random positions of dendritic spines in human cerebral cortex. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(30): Llorens-Martin M, Blazquez-Llorca L, Benavides-Piccione R, Rabano A, Hernandez F, Avila J, et al. Selective alterations of neurons and circuits related to early memory loss in Alzheimer's disease. Front Neuroanat. 2014;8:38. Epub 2014 May 27. Ferreira FR, Nogueira MI, Defelipe J. The influence of James and Darwin on Cajal and his research into the neuron theory and evolution of the nervous system. Front Neuroanat. 2014;8:1. Epub 2014 Jan 29. Lopez-Cruz PL, Larranaga P, DeFelipe J, Bielza C. Bayesian network modeling of the consensus between experts: An application to neuron classification. International Journal of Approximate Reasoning. 2014;55(1):

56 Merchan-Perez A, Rodriguez JR, Gonzalez S, Robles V, Defelipe J, Larranaga P, et al. Three-dimensional spatial distribution of synapses in the neocortex: a dual-beam electron microscopy study. Cerebral cortex (New York, NY: 1991). 2014;24(6): Mellström B, Sahun I, Ruiz-Nuno A, Murtra P, Gomez-Villafuertes R, Savignac M, et al. DREAM controls the on/off switch of specific activity-dependent transcription pathways. Molecular and cellular biology. 2014;34(5): Research Projects Code: Cajal Blue Brain Project Title: Cajal Blue Brain Project. International Blue Brain Proyect Principal Investigator: Javier de Felipe CIBERNED s collaboration: Yes CIBERNED s groups: G403; G204 Type: International Funding Agency: Ecole Polytechnique Federale de Lausanne (Suiza), IBM, Universidad Politecnica de Madrid, Consejo Superior de Investigaciones Científicas, España Funding: N.D. Duration: Code: 2013/07A Title: Papel de GSK-3 β en las alteraciones de los circuitos corticales que ocurren en la enfermedad de Alzheimer Principal Investigator: Teresa Iglesias Vacas CIBERNED s collaboration: Yes CIBERNED s groups: G401; G403; G111; G504; G307 Type: International Funding Agency: CIBERNED Funding: Duration: Program 1 50

57 Group Jesús de Pedro Cuesta Program 1 CONTACT DETAILS Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús Centro de Nacional Pedro Cuesta de Epidemiología Group Jesús de Pedro Cuest Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Instituto Cuesta de Salud Group Carlos Jesús III de Pedro Cuest Monforte de Lemos, 5 Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuest Madrid (Spain) Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Phone: Cuesta +34 Group Jesús 650 de Pedro Cuest Fax: Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuest jpedro@isciii.es Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Group Jesús de Pedro Cuesta Gru LIST OF PERSONNEL Jesús de Pedro Cuesta Principal Investigator, Head of Unit Enrique Alcalde Cabero PhD student Javier Almazán Isla Technician Fuencisla Avellanal Calzadilla Technician Miguel Calero Lara Researcher Lab Head Olga Calero Rueda Postdoctoral Fellow CIBERNED Javier Damián Moreno Postdoctoral Fellow Belén Frades Payo Technician Pablo Martínez Martín Staff Researcher María José Medrano Albero Staff Researcher María del Carmen Rodríguez Blázquez Research Assistant María Ruiz Tovar Postdoctoral Fellow 51

58 Summary The Public Health subgroup reported epidemiological features of Creutzfeldt-Jakob disease (CJD) suggesting that a susceptibility to exposure effects exist determined by juvenile age at exposure. Data for bovine spongiform encephalopathy related diet and routine surgery were implicated with variant CJD and sporadic CJD respectively. Detailed disability prevalence figures were first reported for populations aged > 50 years at Aragon. Patterns for 80 year old with severe limitations in mobility, self-care and domestic activities reveal the subsequent need of help. Increasing detected incidence of multiple sclerosis among Portuguese populations and the value of capture-recapture techniques studies raise concerns about multiple sclerosis incidence trends. New activities were attempted framed on collaboration with IMSERSO (Institute of Aging) related to the new scales for disability assessment. Along 2014, the work on consolidated research lines of the group has continued. Our Group of Research on Quality of Life and Aging followed the activity focused on quality of life in dementia, institutionalized and community-dwelling elderly, and caregiver burden, collaborating with REDYSSEC. In regard to Assessment and Outcomes Research, we can highlight the collaborative study with King s College Hospital, London, UK, on perceptions and expectations in advanced therapies for Parkinson s disease; the descriptive and comparative study on Cameroonian vs. Spanish Parkinson s disease patients; anger in multiple sclerosis; and psychiatric disorders in Parkinson s disease. Most of the studies carried out by this Group were performed in an international setting and some of them were collaborative with CIBERSAM. As Chair of the Committee on Development of Rating Scales of the Movement Disorder Society, we had the opportunity of sharing a relevant research activity on measures and outcomes with the International Group of Restless Legs Syndrome, to be followed in the coming years. Finally, the Non-Motor Symptoms Group, from which Steering Committee we are members, has produced interesting studies on gradation of severity in Parkinson s disease; non-motor symptoms in cervical dystonia; off situation and pain in Parkinson s, and non-motor symptoms in Progressive Supranuclear Palsy. The Molecular Analysis subgroup focused on the analysis of biomarkers and genetic susceptibility markers of different neurodegenerative disorders, including Alzheimer s disease and human prion disorders, as well as in the study of the potential interaction between AD and vascular pathology. These activities have been carried out with the collaboration of different CIBERNED groups, the CIEN Foundation and several international teams, with funding from EU Joint Programme Neurodegenerative Disease Research (JPND), and a grant from the Health Research and Development Strategy. In the 2014 call of grants for Collaborative Projects of CIBERNED, we have been awarded with a grant for the project entitle epigenetic mechanisms involved in the etiology and progression of rapid progressive neurodegenerative dementias, in collaboration with the CIBERNED group from Dr. Isidre Ferrer, José Antonio del Río and José Ángel Berciano (PI for the project Dr. Pascual Sánchez). During this year, the group has also continued performing tasks related to the molecular diagnosis of human prion disease for the Spanish National Health System. Keywords Alzheimer disease, Prion diseases, Creutzfeldt-Jakob, Neurodegenerative disorders, Conformational disorders, Biomarkers, Genetic susceptibility markers, Risk genes, Vascular factors, Aging Program 1 52

59 2014 Annual Report Jesús de Pedro cuesta Group Publications Leon G, Sanchez-Ruiloba L, Perez-Rodriguez A, Gragera T, Martinez N, Hernandez S, et al. Shoc2/Sur8 protein regulates neurite outgrowth. PloS one. 2014;9(12):e Epub 2014 Dec 16. Garatachea N, Emanuele E, Calero M, Fuku N, Arai Y, Abe Y, et al. ApoE gene and exceptional longevity: Insights from three independent cohorts. Exp Gerontol. 2014;53: Rabano A, Rodal I, Cuadros R, Calero M, Hernandez F, Avila J. Argyrophilic grain pathology as a natural model of tau propagation. Journal of Alzheimer's disease: JAD. 2014;40(Suppl 1):S Garcia Lopez F. Cual es el efecto de los quelantes de fósforo basados en calcio en comparación con los quelantes sin calcio en la mortalidad de los pacientes con enfermedad renal crónica? Análisis crítico de Jamal SA, Vandermeer B, Raggi P, et al. Effect of calcium-base. NefroPlus. 2014;6(1):25-8. Garcia Lopez F, Delgado-Cordova M. Cual es el efecto de un aumento en el consumo de potasio sobre la presión arterial y sobre la morbilidad y mortalidad cardiovascular y cuales son sus efectos adversos en adultos y ninos aparentemente sanos? Análisis crítico de Aburto NJ, Hanson S, Gutier. NefroPlus. 2014;6(1):21-4. Garcia Lopez F, Gallegos-Villalobos A. Cual es el efecto sobre la salud de un consumo bajo de sodio? Análisis crítico de Aburto NJ, Ziolkovska A, Hooper L, et al. Effect of lower sodium intake on health: systematic review and meta-analyses. BMJ. 2013;346:f1326. NefroPlus. 2014;6(1):44-6. Martinez-Martin P. Cuánto ha mejorado la calidad de vida de los pacientes con enfermedad de Parkinson? Rev Esp Trast Mov. 2014;6: Lovestone S, Boada M, Dubois B, Hüll M, Rinne JO, Huppertz HJ, et al. A Phase II Trial of Tideglusib in Alzheimer's Disease. Journal of Alzheimer's disease: JAD. [Epub ahead of print]. Epub 2014 Dec 23. Di Gioia MC, Gallar Ruiz P, Cobo G, Garcia Lopez F, Agud Aparicio JL, Oliet A, et al. Body composition changes in hemodialysis patients: implications for prognosis. Nephrology and Renal Studies. 2014;1(1):1-7. Zea-Sevilla MA, Bermejo-Velasco P, Serrano-Heranz R, Calero M. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) associated with a novel C82R mutation in the NOTCH3 gene. Journal of Alzheimer's disease: JAD. 2015;43(2): Epub Castro-Monteiro E, Forjaz MJ, Ayala A, Rodriguez-Blazquez C, Fernandez-Mayoralas G, Diaz-Redondo A, et al. Change and predictors of quality of life in institutionalized older adults with dementia. Quality of life research: an international journal of quality of life aspects of treatment, care and rehabilitation. 2014;23(9): Epub 2014 May 7. Rizos A, Martinez-Martin P, Odin P, Antonini A, Kessel B, Kozul TK, et al.; EUROPAR and the IPDMDS non-motor PD Study Group. Characterizing motor and non-motor aspects of early-morning off periods in Parkinson's disease: an international multicenter study. Parkinsonism & related disorders. 2014;20(11): Epub 2014 Sep 16. Reichmann H, Martinez-Martin P, Stocchi F. Effect of therapeutic interventions on health-related quality of life in Parkinson s Disease. Eur Neurol Rev. 2014;9: Granado-Lorencio F, Lagarda MJ, Garcia-Lopez FJ, Sanchez-Siles LM, Blanco-Navarro I, Alegria A, et al. Effect of β-cryptoxanthin plus phytosterols on cardiovascular risk and bone turnover markers in post-menopausal women: a randomized crossover trial. Nutrition, metabolism, and cardiovascular diseases: NMCD. 2014;24(10): Epub 2014 May 6. Martinez-Martin P, Rodriguez-Blazquez C, Forjaz MJ, Ray Chaudhuri K. Guide to Assessment Scales in Parkinson s Disease van der Heeden JF, Marinus J, Martinez-Martin P, van Hilten JJ. Importance of nondopaminergic features in evaluating disease severity of Parkinson disease. Neurology. 2014;82(5): Epub 2014 Jan 3. de Sa J, Alcalde-Cabero E, Almazan-Isla J, Lopez F, de Pedro-Cuesta J. Incidence of multiple sclerosis in Northern Lisbon, Portugal: BMC neurology. 2014;14(1):249. Epub 2014 Dec

60 Rodriguez-Violante M, Cervantes-Arriaga A, Velazquez-Osuna S, Llorens-Arenas R, Calderon-Fajardo H, Pina-Fuentes D, et al. Independent Validation of the SEND-PD and Correlation with the MDS- UPDRS Part IA. Parkinson's disease. 2014;2014: Epub 2014 Oct 23. Rojo-Perez F, Fernandez-Mayoralas G, Forjaz MJ, Prieto-Flores ME, Martinez-Martin P. La influencia de las condiciones del entorno residencial y personal en la calidad de vida de los adultos mayores españoles Olazaran J, Gonzalez B, Osa-Ruiz E, Felipe-Ruiz S, Boyano I, Fontani V, et al. Motor effects of radio electric asymmetric conveyer in Alzheimer's disease: results from a cross-over trial. Journal of Alzheimer's disease: JAD. 2014;42(1): Marventano S, Ayala A, Gonzalez N, Rodriguez-Blazquez C, Garcia-Gutierrez S, Forjaz MJ; Spanish Research Group of Quality of Life and Ageing. Multimorbidity and functional status in communitydwelling older adults. European journal of internal medicine. 2014;25(7): Epub 2014 Jul 3. Benito-Leon J, Labiano-Fontcuberta A, Mitchell AJ, Moreno-Garcia S, Martinez-Martin P. Multiple sclerosis is associated with high trait anger: a case-control study. Journal of the neurological sciences. 2014;340(1-2): Epub 2014 Feb 26. Klingelhoefer L, Martino D, Martinez-Martin P, Sauerbier A, Rizos A, Jost W,Warner TT, et al. Non motor symptoms and focal cervical dystonia: observations from 102 patients. Basal Ganglia. 2014;4: Garcia-Ruiz PJ, Chaudhuri KR, Martinez-Martin P. Non-motor symptoms of Parkinson's disease A review from the past. Journal of the neurological sciences. 2014;338(1-2):30-3. Epub 2014 Jan 8. Zis P, Rizos A, Martinez-Martin P, Pal S, Silverdale M, Sharma JC, et al. Non-motor symptoms profile and burden in drug naïve versus long-term Parkinson's disease patients. Journal of Parkinson's disease. 2014;4(3): Reddy P, Martinez-Martin P, Brown RG, Chaudhuri KR, Lin JP, Selway R, et al. Perceptions of symptoms and expectations of advanced therapy for Parkinson's disease: preliminary report of a Patient-Reported Outcome tool for Advanced Parkinson's disease (PRO-APD). Health and quality of life outcomes. 2014;12:11. Epub 2014 Jan 24. Borchani H, Bielza C, Martinez-Martin P, Larranaga P. Predicting the EQ-5D from the Parkinson's disease questionnaire PDQ-8 using multi-dimensional Bayesian network classifiers. Biomed Engineering. 2014;26(1): Stocchi F, Martinez-Martin P, Reichmann H. Quality of life in Parkinson s Disease Patient, clinical, and research perspectives. Eur Neurol Rev. 2014;9:12-8. Zea-Sevilla MA, Martinez-Martin P. Rating scales and questionnaires for assessment of sleep disorders in Parkinson's disease: what they inform about? Journal of neural transmission. 2014;121(Suppl 1):S Epub 2014 Apr 23. Walters AS, Frauscher B, Allen R, Benes H, Chaudhuri KR, Garcia-Borreguero D, et al.; members of the MDS Committee on Rating Scales. Review of Diagnostic Instruments for the Restless Legs Syndrome/ Willis-Ekbom Disease (RLS/WED): Critique and Recommendations. Journal of clinical sleep medicine: JCSM: official publication of the American Academy of Sleep Medicine. 2014;10(12): Epub 2014 Dec 15. Walters AS, Frauscher B, Allen R, Benes H, Chaudhuri KR, Garcia-Borreguero D, et al.; Members of the MDS Committee on Rating Scales. Review of Quality of Life Instruments for the Restless Legs Syndrome/Willis-Ekbom Disease (RLS/WED): Critique and Recommendations. Journal of clinical sleep medicine: JCSM: official publication of the American Academy of Sleep Medicine. 2014;10(12): Epub 2014 Dec 15. Walters AS, Frauscher B, Allen R, Benes H, Ray Chaudhuri K, Garcia-Borreguero D, et al.; Members of the MDS Committee on Rating Scales. Review of Severity Rating Scales for the Restless Legs Syndrome: Critique and recommendations. Mov Disord Clin Pract. 2014;1: Martinez-Martin P, Rodriguez-Blazquez C, Kurtis MM. Scales and tests in Movement Disorders. In: Wolters E, Baumann C, Ed. Parkinson s Disease and Other Movement Disorders Rodriguez-Blazquez C, Sauerbier A, Ray Chaudhuri K, Martinez-Martin P. Scales for Measuring Parkinsonism in Demented Patients. In: Merello M, Starktstein SE, Ed Pino L, Gonzalez-Velez AE, Prieto-Flores ME, Ayala A, Fernandez-Mayoralas G, Rojo-Perez F, et al. Self- Program 1 54

61 2014 Annual Report Jesús de Pedro cuesta Group perceived health and quality of life by activity status in community-dwelling older adults. Geriatrics & gerontology international. 2014;14(2): Epub 2013 Jul 26. Giglio RE, Rodriguez-Blazquez C, de Pedro-Cuesta J, Forjaz MJ. Sense of coherence and health of community-dwelling older adults in Spain. International psychogeriatrics / IPA. [Epub ahead of print]1-8. Epub 2014 Nov 25. Rivera-Navarro J, Cubo E, Almazan J. The Impact of Tourette s Syndrome in the School and the Family: Perspectives from Three Stakeholder Groups. International Journal for the Advancement of Counselling. 2014;36(1): Leentjens AF, Dujardin K, Pontone GM, Starkstein SE, Weintraub D, Martinez-Martin P. The Parkinson Anxiety Scale (PAS): development and validation of a new anxiety scale. Movement disorders: official journal of the Movement Disorder Society. 2014;29(8): Epub 2014 May 23. de Pedro-Cuesta J, Mahillo-Fernandez I, Calero M, Rabano A, Cruz M, Siden Å, et al.; EUROSURGYCJD Research Group. Towards an age-dependent transmission model of acquired and sporadic Creutzfeldt-Jakob disease. PloS one. 2014;9(10):e Epub 2014 Oct 3. Gallegos-Villalobos A, Garcia-Lopez F, Escalada C, Ortiz JJ, Cardona J, Medina A, et al. Use of radioactive iodine I-131 and monitoring of radioactivity in patients with chronic kidney disease on haemodialysis. Nefrologia: publicación oficial de la Sociedad Española Nefrología. 2014;34(3): Epub 2014 Feb 10. Forjaz MJ, Rodriguez-Blazquez C. Spain, Personal Well-Being Index. Validation with Older Adults. In: Michalos AC, Ed. Encyclopedia of Quality of Life and Well-Being Research Drutyte G, Forjaz MJ, Rodriguez-Blazquez C, Martinez-Martin P, Breen KC. What impacts on the stress symptoms of Parkinson's carers?: Results from the Parkinson's UK Members' Survey. Disabil Rehabil. 2014;36: Navarro-Gil P, Gonzalez-Velez AE, Ayala A, Martin-Garcia S, Martinez-Martin P, Forjaz MJ; Spanish Research Group on Quality of Life and Ageing. Which factors are associated with mortality in institutionalized older adults with dementia? Archives of gerontology and geriatrics. 2014;59(3): Epub 2014 Jul 14. Granado-Lorencio F, de Las Heras L, Millan CS, Garcia-Lopez FJ, Blanco-Navarro I, Perez-Sacristan B, et al. β-cryptoxanthin modulates the response to phytosterols in post-menopausal women carrying NPC1L1 L272L and ABCG8 A632 V polymorphisms: an exploratory study. Genes & nutrition. 2014;9(5):428. Epub 2014 Aug 28. Diaz-Redondo A, Rodriguez-Blazquez C, Ayala A, Martinez-Martin P, Forjaz MJ; Spanish Research Group on Quality of Life and Aging. EQ-5D rated by proxy in institutionalized older adults with dementia: psychometric pros and cons. Geriatrics & gerontology international. 2014;14(2): Martinez-Martin P, Hernandez B, Ricart J; FAST Study Group. Factors determining when to start levodopa/carbidopa/entacapone treatment in Spanish patients with Parkinson's disease. Neurologia (Barcelona, Spain). 2014;29(3): Starkstein SE, Dragovic M, Dujardin K, Marsh L, Martinez-Martin P, Pontone GM, et al. Anxiety has specific syndromal profiles in Parkinson disease: a data-driven approach. The American journal of geriatric psychiatry: official journal of the American Association for Geriatric Psychiatry. 2014;22(12): Martinez-Martin P. Nonmotor symptoms and health-related quality of life in early Parkinson's disease. Movement disorders: official journal of the Movement Disorder Society. 2014;29(2): Dodel R, Jönsson B, Reese JP, Winter Y, Martinez-Martin P, Holloway R, et al. Measurement of costs and scales for outcome evaluation in health economic studies of Parkinson's disease. Movement disorders: official journal of the Movement Disorder Society. 2014;29(2): Almazan-Isla J, Comin-Comin M, Damian J, Alcalde-Cabero E, Ruiz C, Franco E, et al. Analysis of disability using WHODAS 2.0 among the middle-aged and elderly in Cinco Villas, Spain. Disability and health journal. 2014;7(1): Martinez-Martin P, Rodriguez-Blazquez C, Forjaz MJ, Alvarez-Sanchez M, Arakaki T, Bergareche-Yarza A, et al. Relationship between the MDS-UPDRS domains and the health-related quality of life of Parkinson's disease patients. European journal of neurology: the official journal of the European 55

62 Federation of Neurological Societies. 2014;21(3): Rivera-Navarro J, Cubo E, Almazan J. The Impact of Tourette s Syndrome in the School and the Family: Perspectives from Three Stakeholder Groups. International Journal for the Advancement of Counselling. 2014;36(1): Calero M,Gutierrez L, Salas G, Luengo Y, Lazaro A, Acedo P, et al. Efficient and safe internalization of magnetic iron oxide nanoparticles: two fundamental requirements for biomedical applications. Nanomedicine: nanotechnology, biology, and medicine. 2014;10(4): Research Projects Code: Fundación Ramón Areces 2012 Title: Estudio de la capacidad neuroprotectora de las células epiteliales de los plexos coroideos como potencial terapia regenerativa de la enfermedad de Alzheimer Principal Investigator: Eva Carro CIBERNED s collaboration: Yes CIBERNED s groups: G509; G502 Type: National Funding Agency: Fundación Ramón Areces Funding: Duration: Code: DEMTEST-MPY 990/12 Title: Biomarker based diagnosis of rapid progressive dementias - optimisation of diagnostic protocols (DemTest) Principal Investigator: M Calero CIBERNED s groups: Type: International Funding Agency: Unión Europea FIS. Joint Programming Neurodegenerative Disease Call for transnational research proposals 2011 "Neurodegenerative Diseases Funding: Duration: Program 1 56

63 Group Isidro Ferrer Abizanda Program 1 CONTACT DETAILS Group Isidro Ferrer Abizanda Group Isidro Ferrer Institut Abizanda de Neuropatologia Group Isidro Servei Ferrer Abizanda d Anatomia Group Patológica Isidro Ferrer Abizand Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda IDIBELL-Hospital Group Isidro Universitari Ferrer Abizanda de Bellvitge Group Isidro Ferre Carrer Feixa LLarga sn Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidr Hospitalet de Llobregat (Spain) Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Phone: Abizanda Group 452 / Isidro Ferrer 808 Abizanda Group Fax: Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Grou 8082ifa@gmail.com Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizanda Group Isidro Ferrer Abizand LIST OF PERSONNEL Isidro Ferrer Abizanda Principal Investigator Marta Barrachina Castillo Research staff Ester Aso Research staff Karina Hernández Ortega Research staff Montserrat Olivé Plana Research staff Xavier Altafaj Research staff Mercedes Armand-Ugon Research staff Irene López González PhD student Paula García-Esparcia PhD student Belén Ansoleaga Ávila PhD student Mayelin González PhD student Anusha Koneti PhD student Margarita Carmona Murillo Technician Rosa Blanco Soto Technician María Dolores Moreno León Technician Jesús Moreno Castro Technician Lourdes Castillo Valentín Technician María José Zújar Rubio Technician Sara Berjawi Student Pol Véneto Andrés Student Luis Alberto Escobar Research visitor 57

64 Summary The main focus during the last year has been the use of combined -omics and robust bioinformatics processing of obtained data to identify altered transcriptomics, proteomics and metabolomics in selected brain regions at early stages of the principal neurodegenerative diseases including Alzheimer s disease, Parkinson s disease, tauopathies and Creutzfeld-Jakob s disease and corresponding animal models. Major achievements include the identification of i) early neuroinflammatory deregulation, ii) confluent altered protein synthesis pathways, iii) altered energy metabolism, and iv) altered purine metabolism, all of which are region-, stage-, disease- and species-dependent, and which involve complex responses. These alterations are expressed as particular metabolic abnormalities which can be developed into potential biomarkers. These alterations also reveal potential targets of therapeutic intervention preceding by several years the appearance of clinical symptoms. This applies to human individuals aged between 55 and 70 years. In addition, a number of particular aspects of other diseases have been examined as collateral derivations of the principal lines covering amyotrophic lateral sclerosis and Huntington s disease. Some results have been translated to animal models to verify commonly altered pathways and to design therapeutic assessments; and treatments with particular molecules such as new generation glycodendrimers, cannabinoids, carboxy-erythropoietin and new synthetic compounds have been successfully assayed in mouse models. Yet inter-species differences are reminders of the need for caution in the direct application to human beings of drugs that are beneficial in rodents. Our experience and that of many others shows that rodents are particularly responsive to different drugs that later on do not work in human cases. This scenario strengths the importance of the study of human brain samples to understand diseases which are specific or selective of human beings. Even considering these limitations, a pilot study in amyotrophic lateral sclerosis is ongoing, and another one in individuals with mild cognitive impairment is in preparation. Regarding biomarkers, a series of studies have been carried out to improve understanding of the relationship between certain biomarkers in the CSF and the real situation in the brain in the same group of cases. This approach has been useful in differential diagnosis in the group of rapid dementias. About thirty papers have appeared, most of them in collaboration with other groups of CIBERNED and other CIBERs, and with international teams from a good number of countries. Five doctoral theses are in course, and one patent has been produced. The main strength of the group is the molecular study of brain modifications with aging and pathological states with disease progression using optimal post-mortem brain samples from the Institute of Neuropathology Brain Bank (a branch of the HUB-ICO-IDIBELL Biobank that belongs to the RETICS Biobank, Institute of Health Carlos III) properly processed after several years of experience. These skills make the group an international reference in the field of advanced human neuropathology. Keywords Neuropathology, Epigenetics, Transcriptomics, Proteomics, Metabolomics, Lipidomics, Alzheimer s, Parkinson s, Creutzfeldt-Jakob disease, Frontotemporal lobar degeneration, Amyotrophic lateral sclerosis, Chromosome X-linked adrenoleukodystrophy, Muscle diseases Program 1 58

65 2014 Annual Report Isidro Ferrer Abizanda Group Publications Vergara C, Ordonez-Gutierrez L, Wandosell F, Ferrer I, Del Rio JA, Gavin R. Role of PrPC Expression in Tau Protein Levels and Phosphorylation in Alzheimer's Disease Evolution. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Jun 26. Llorens F, Zafar S, Ansoleaga B, Shafiq M, Blanco R, Carmona M, et al. Subtype and regional regulation of prion biomarkers in sporadic Creutzfeldt-Jakob disease. Neuropathology and applied neurobiology. [Epub ahead of print]. Epub 2014 Aug 18. Porquet D, Grinan-Ferre C, Ferrer I, Camins A, Sanfeliu C, Del Valle J, et al. Neuroprotective role of trans-resveratrol in a murine model of familial Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;42(4): Pfeffer G, Sambuughin N, Olive M, Tyndel F, Toro C, Goldfarb LG, et al. A new disease allele for the p.c30071r mutation in titin causing hereditary myopathy with early respiratory failure. Neuromuscular disorders: NMD. 2014;24(3): Epub 2013 Dec 11. Böhm J, Biancalana V, Malfatti E, Dondaine N, Koch C, Vasli N, et al. Adult-onset autosomal dominant centronuclear myopathy due to BIN1 mutations. Brain: a journal of neurology. 2014;137(Pt 12): Epub 2014 Sep 25. Fabelo N, Martin V, Marin R, Moreno D, Ferrer I, Diaz M. Altered lipid composition in cortical lipid rafts occurs at early stages of sporadic Alzheimer's disease and facilitates APP/BACE1 interactions. Neurobiology of aging. 2014;35(8): Epub 2014 Feb 11. Varani AP, Aso E, Moutinho LM, Maldonado R, Balerio GN. Attenuation by baclofen of nicotine rewarding properties and nicotine withdrawal manifestations. Psychopharmacologia. 2014;231(15): Epub 2014 Feb 20. Launay N, Aguado C, Fourcade S, Ruiz M, Grau L, Riera J, et al. Autophagy induction halts axonal degeneration in a mouse model of X-adrenoleukodystrophy. Acta neuropathologica. [Epub ahead of print]. Epub 2014 Dec 31. Fernandez-Echevarria C, Diaz M, Ferrer I, Canerina-Amaro A, Marin R. Aβ promotes VDAC1 channel dephosphorylation in neuronal lipid rafts. Relevance to the mechanisms of neurotoxicity in Alzheimer's disease. Neuroscience. 2014;278: Epub 2014 Aug 26. Varani AP, Aso E, Maldonado R, Balerio GN. Baclofen and 2-hydroxysaclofen modify acute hypolocomotive and antinociceptive effects of nicotine. European journal of pharmacology. 2014;738: Epub 2014 Jun 2. Aso E, Ferrer I. Cannabinoids for treatment of Alzheimer's disease: moving toward the clinic. Frontiers in pharmacology. 2014;5:37. Epub 2014 Mar 5. Aso E, Sanchez-Pla A, Vegas-Lozano E, Maldonado R, Ferrer I. Cannabis-Based Medicine Reduces Multiple Pathological Processes in AβPP/PS1 Mice. Journal of Alzheimer's disease: JAD. 2015; 43(3): Epub Garcia-Esparcia P, Llorens F, Carmona M, Ferrer I. Complex deregulation and expression of cytokines and mediators of the immune response in Parkinson's disease brain is region dependent. Brain pathology (Zurich, Switzerland). 2014;24(6): Epub 2014 Apr 14. Parra-Damas A, Valero J, Chen M, Espana J, Martin E, Ferrer I, et al. Crtc1 activates a transcriptional program deregulated at early Alzheimer's disease-related stages. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(17): Ansoleaga B, Garcia-Esparcia P, Pinacho R, Haro JM, Ramos B, Ferrer I. Decrease in olfactory and taste receptor expression in the dorsolateral prefrontal cortex in chronic schizophrenia. Journal of psychiatric research. 2015;60: Epub 2014 Oct 2. Cacabelos D, Ayala V, Ramirez-Nunez O, Granado-Serrano AB, Boada J, Serrano JC, et al. Dietary lipid unsaturation influences survival and oxidative modifications of an amyotrophic lateral sclerosis model in a gender-specific manner. Neuromolecular medicine. 2014;16(4): Epub 2014 Jul 1. Hoegg-Beiler MB, Sirisi S, Orozco IJ, Ferrer I, Hohensee S, Auberson M, et al. Disrupting MLC1 and 59

66 GlialCAM and ClC-2 interactions in leukodystrophy entails glial chloride channel dysfunction. Nature communications. 2014;5:3475. Epub 2014 Mar 19. Grau C, Arato K, Fernandez-Fernandez JM, Valderrama A, Sindreu C, Fillat C, et al. DYRK1A-mediated phosphorylation of GluN2A at Ser(1048) regulates the surface expression and channel activity of GluN1/GluN2A receptors. Frontiers in cellular neuroscience. 2014;8:331. Epub 2014 Oct 17. Sepulveda-Falla D, Barrera-Ocampo A, Hagel C, Korwitz A, Vinueza-Veloz MF, Zhou K, et al. Familial Alzheimer's disease-associated presenilin-1 alters cerebellar activity and calcium homeostasis. The Journal of clinical investigation. 2014;124(4): Epub 2014 Feb 24. Ferrer I, Lopez-Gonzalez I, Carmona M, Arregui L, Dalfo E, Torrejon-Escribano B, et al. Glial and neuronal tau pathology in tauopathies: characterization of disease-specific phenotypes and tau pathology progression. Journal of neuropathology and experimental neurology. 2014;73(1): Fernandez-Nogales M, Cabrera JR, Santos-Galindo M, Hoozemans JJ, Ferrer I, Rozemuller AJ, et al. Huntington's disease is a four-repeat tauopathy with tau nuclear rods. Nature medicine. 2014;20(8): Epub 2014 Jul 20. Pinacho R, Valdizan EM, Pilar-Cuellar F, Prades R, Tarrago T, Haro JM, et al. Increased SP4 and SP1 transcription factor expression in the postmortem hippocampus of chronic schizophrenia. Journal of psychiatric research. 2014;58: Epub 2014 Aug 19. Villar-Menendez I, Porta S, Buira SP, Pereira-Veiga T, Diaz-Sanchez S, Albasanz JL, et al. Increased striatal adenosine A2A receptor levels is an early event in Parkinson's disease-related pathology and it is potentially regulated by mir-34b. Neurobiology of disease. 2014;69: Epub 2014 Jun 2. Kang T, Kim JH, Hong I, Park NH, Heinsen H, Lee JY, et al. Large-scale analysis of posttranslational modifications in the hippocampus of patients with Alzheimer's disease using pi shift and label-free quantification without enrichment. Analytical and bioanalytical chemistry. 2014;406(22): Epub 2014 Aug 15. Armand-Ugon M, Aso E, Moreno J, Riera-Codina M, Sanchez A, Vegas E, et al. Memory Improvement in the AβPP/PS1 Mouse Model of Familial Alzheimer's Disease Induced by Carbamylated-Erythropoietin is Accompanied by Modulation of Synaptic Genes. Journal of Alzheimer's disease: JAD. [Epub ahead of print]. Epub 2014 Dec 29. Grison A, Zucchelli S, Urzì A, Zamparo I, Lazarevic D, Pascarella G, et al. Mesencephalic dopaminergic neurons express a repertoire of olfactory receptors and respond to odorant-like molecules. BMC genomics. 2014;15:729. Epub 2014 Aug 27. Jove M, Portero-Otin M, Naudi A, Ferrer I, Pamplona R. Metabolomics of human brain aging and age-related neurodegenerative diseases. Journal of neuropathology and experimental neurology. 2014;73(7): Benseny-Cases N, Klementieva O, Cotte M, Ferrer I, Cladera J. Microspectroscopy (μftir) Reveals Colocalization of Lipid Oxidation and Amyloid Plaques in Human Alzheimer Disease Brains. Analytical chemistry. 2014;86(24): Epub 2014 Dec 4. Fourcade S, Lopez-Erauskin J, Ruiz M, Ferrer I, Pujol A. Mitochondrial dysfunction and oxidative damage cooperatively fuel axonal degeneration in X-linked adrenoleukodystrophy. Biochimie. 2014;98: Epub 2013 Sep 24. Gerschütz A, Heinsen H, Grünblatt E, Wagner AK, Bartl J, Meissner C, et al. Neuron-specific alterations in signal transduction pathways associated with Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;40(1): Alafuzoff I, Pikkarainen M, Neumann M, Arzberger T, Al-Sarraj S, Bodi I, et al. Neuropathological assessments of the pathology in frontotemporal lobar degeneration with TDP43-positive inclusions: an inter-laboratory study by the BrainNet Europe consortium. Journal of neural transmission. [Epub ahead of print]. Epub 2014 Sep 20. Cabello J, Sämann J, Gomez-Orte E, Erazo T, Coppa A, Pujol A, et al. PDR-1/hParkin negatively regulates the phagocytosis of apoptotic cell corpses in Caenorhabditis elegans. Cell death & disease. 2014;5:e1120. Epub 2014 Mar 13. Sanchez-Mut JV, Aso E, Heyn H, Matsuda T, Bock C, Ferrer I, et al. Promoter hypermethylation of the phosphatase DUSP22 mediates PKA-dependent TAU phosphorylation and CREB activation in Program 1 60

67 2014 Annual Report Isidro Ferrer Abizanda Group Alzheimer's disease. Hippocampus. 2014;24(4): Epub 2014 Jan 28. Villar-Menendez I, Diaz-Sanchez S, Blanch M, Albasanz JL, Pereira-Veiga T, Monje A, et al. Reduced striatal adenosine A2A receptor levels define a molecular subgroup in schizophrenia. Journal of psychiatric research. 2014;51: Epub 2014 Jan 7. Ferrer I. Selection of controls in the study of human neurodegenerative diseases in old age. Journal of neural transmission. [Epub ahead of print]. Epub 2014 Aug 12. Villar-Menendez I, Nunez F, Diaz-Sanchez S, Albasanz JL, Taura J, Fernandez-Duenas V, et al. Striatal adenosine A2A receptor expression is controlled by S-adenosyl-L-methionine-mediated methylation. Purinergic signalling. 2014;10(3): Epub 2014 Jun 19. Llorens F, Lopez-Gonzalez I, Thüne K, Carmona M, Zafar S, Andreoletti O, et al. Subtype and regionalspecific neuroinflammation in sporadic creutzfeldt-jakob disease. Frontiers in aging neuroscience. 2014;6:198. Epub 2014 Aug 4. Kalko SG, Paco S, Jou C, Rodriguez MA, Meznaric M, Rogac M, et al. Transcriptomic profiling of TK2 deficient human skeletal muscle suggests a role for the p53 signalling pathway and identifies growth and differentiation factor-15 as a potential novel biomarker for mitochondrial myopathies. BMC genomics. 2014;15:91. Epub 2014 Feb 1. Ferrer I, Mohan P, Chen H, Castellsague J, Gomez-Baldo L, Carmona M, et al. Tubers from patients with tuberous sclerosis complex are characterized by changes in microtubule biology through ROCK2 signalling. The Journal of pathology. 2014;233(3): Epub 2014 Apr 29. Manuel I, Gonzalez de San Roman E, Giralt MT, Ferrer I, Rodriguez-Puertas R. Type-1 cannabinoid receptor activity during Alzheimer's disease progression. Journal of Alzheimer's disease: JAD. 2014;42(3): Lopez-Gonzalez I, Carmona M, Arregui L, Kovacs GG, Ferrer I. αb-crystallin and HSP27 in glial cells in tauopathies. Neuropathology: official journal of the Japanese Society of Neuropathology. 2014;34(6): Epub 2014 Jul 2. Kalia LV, Lang AE, Hazrati LN, Fujioka S, Wszolek ZK, Dickson DW, et al. Clinical Correlations With Lewy Body Pathology in LRRK2-Related Parkinson Disease. JAMA neurology. 2015;72(1): Epub Gomez-Suaga P, Rivero-Rios P, Fdez E, Blanca Ramirez M, Ferrer I, Aiastui A, et al. LRRK2 delays degradative receptor trafficking by impeding late endosomal budding through decreasing Rab7 activity. Human molecular genetics. 2014;23(25): Epub 2014 Jul 30. Llorens F,Ferrer I,del Rio JA. Gene expression resulting from PrPC ablation and PrPC overexpression in murine and cellular models. Molecular neurobiology. 2014; 49(1): Sanmillan JL, Plans G, Vidal N, Acebes JJ. Supratentorial brain schwannomas: an uncommon location for a common tumour. British journal of neurosurgery. 2014;28(1):25-8. Suarez-Calvet M, Dols-Icardo O, Llado A, Sanchez-Valle R, Hernandez I, Amer G, et al. Plasma phosphorylated TDP-43 levels are elevated in patients with frontotemporal dementia carrying a C9orf72 repeat expansion or a GRN mutation. Journal of neurology, neurosurgery, and psychiatry. 2014;85(6): Robledo P, Martin-Garcia E, Aso E, Maldonado R. Genetically modified mice as tools to understand the neurobiological substrates of depression. Current pharmaceutical design. 2014;20(23): Research Projects Code: CD10/00121 Title: Ayudas para contratos postdoctorales de perfeccionamiento Sara Borrell Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: National Funding Agency: Instituto Carlos III Funding: Duration: Code: FI12/00410 Title: Ayudas Predoctorales de formación en investiga- 61

68 ción (PFIS) Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: National Funding Agency: FIS - Instituto de Salud Carlos III Referencia Funding: Duration: Code: CAD-BIO-04 Title: Banc Teixits Neurològics de Catalunya Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: Others Funding Agency: Farmaindustria Funding: Duration: Code: XVII Premi àmbit de la gent gran Title: Cannabinoides y Alzheimer: potencial efecto terapéutico en un modelo animal de la enfermedad Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: National Funding Agency: Fundació Agrupació Mutua Funding: Duration: Code: Title: DEVELAGE Principal Investigator: Ferrer I. Medizinische Universität Wien CIBERNED s groups: Type: International Funding Agency: Comisión Europea Funding: Duration: Code: PI Title: Estudio clínico, patológico y genético en miopatías cardioesqueléticas hereditarias Principal Investigator: Montserrat Olive Plana CIBERNED s groups: Type: National Funding Agency: Instituto de Salud Carlos III Funding: ,88 Duration: Code: PI Title: Identificación de un transcriptoma, proteoma y lipidoma de riesgo en los primeros estadios de la enfermedad de Parkinson Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: National Funding Agency: Instituto Carlos III, Fondo de Investigación Sanitaria Funding: Duration: Code: Althia-Fundació La Caixa Conveni Col Laboració Title: Identificació d'un model predictiu d'opció terapèutica en gbm mitjançant l'aplicació de tècniques de patologia de sistemes. Extrapolació d'anàlisi a altres tipus de tumors Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: CCAA Funding Agency: Althia - Fundació La Caixa Funding: Duration: Code: PT13/0010/0013 Title: Plataforma de Biobancos Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: International Funding Agency: FIS - Instituto de Salud Carlos III Funding: Duration: 2014 Code: BFI Title: Program de becas para la formación del personal investigador (modalidad AE) Principal Investigator: Isidro Ferrer CIBERNED s groups: Type: International Funding Agency: Gobierno Vasco Funding: Duration: 2014 Code: RD09/0076/00056 Title: RETIC Biobancos Principal Investigator: Isidro Ferrer Program 1 62

69 2014 Annual Report Isidro Ferrer Abizanda Group CIBERNED s groups: Type: National Funding Agency: Instituto de Salud Carlos III Funding: N.D. Duration: 2014 Code: PI11/03028 Title: DEMTEST: Biomarker based diagnosis of rapid progressive dementias Principal Investigator: Pascual Sánchez CIBERNED s collaboration: Yes CIBERNED s groups: G601; G114; G503; G609 Type: Others Funding Agency: Instituto de Salud Carlos III - Comunidad Económica Europea Funding: Duration:

70 Group Antonia Gutiérrez Pérez Program 1 CONTACT DETAILS oup Antonia Gutiérrez Pérez Group Dpto. Antonia Biología Gutiérrez Celular, Pérez Genética Group y Fisiología Antonia Gutiérrez (Área de Biología Pérez Group Celular) Antonia Gutiérrez rez Group Antonia Gutiérrez Pérez Group Antonia Gutiérrez Facultad Pérez Group de Ciencias, Antonia Universidad Gutiérrez de Pérez Málaga Group Antonia Campus de Teatinos tiérrez Pérez Group Antonia Gutiérrez Pérez Group Antonia Gutiérrez Pérez Group Antonia Málaga Gutiérrez (Spain) Pérez Group agutierrez@uma.es tonia Gutiérrez Pérez Group Antonia Gutiérrez Pérez Group Antonia Gutiérrez Pérez Group Antonia Gutiérrez Pérez Phone: oup Antonia Gutiérrez Pérez Group Antonia Gutiérrez Pérez Group Antonia Gutiérrez Pérez Group Antonia Gutier LIST OF PERSONNEL Antonia Gutiérrez Pérez Principal Investigator, Full Professor José Carlos Dávila Cansino Full Professor Zafaruddin Khan Lecturer Manuel Romero Acebal Chief of Neurology Department, Assistant Professor David Baglietto Vargas Postdoctoral Fellow Inés Moreno González Postdoctoral Fellow Raquel Sánchez Varo CIBERNED Postdoctoral Researcher Elisabeth Sánchez Mejías PhD student Laura Trujillo Estrada PhD student Vanessa De Castro Carratalá CIBERNED Staff Researcher Mercedes Aneiros Ferrer CIBERNED Staff Researcher 64

71 2014 Annual Report Antonia Gutiérrez Pérez Group Summary During 2014 we have continued with our study on the inflammatory response (microglial activation) in the hippocampus of patients with Alzheimer s disease (AD), from initial stages of pathology (Braak II) to advanced ones (Braak V-VI). The expression of specific markers of microglial activation/differentiation has been analyzed by immunohistochemical, stereological and image analysis techniques, in parallel with biochemical/molecular techniques in collaboration with the group of Dr. J. Vitorica (University of Sevilla). Results show that the microglial response in patients is in clear contrast with the observed in PS1/APP transgenic models. Therefore, the current models of AD, based in Abeta production, do not reproduce the microglial pathology seen in AD patients, which might explain the failure obtained in translating anti-inflammatory therapies from the preclinical studies in models to the human clinic. Furthermore, and in collaboration with Dr. J. Comella (Institut of Recerca, Hospital Vall d Hebron), we have analyzed the expression of FAIM-L in the hippocampus and entorrinal cortex of PS1/APP models. There is a significant decrease in FAIM-L expression in the models as in the brains of AD patients, and this reduction is associated with the progression of the neurodegeneration by changing the inflammatory response mediated by TNFalfa in neurons. This research on the inflammatory process in patients and models was integrated within the aims of the collaborative CIBERNED project that we initiated in Continuing with this research line, we are now trying to determine the role of the microglial activation, by pharmacological or genetic manipulation of APP or PS1/APP models, on the progression of the pathology (Abeta plaques, phospho-tau, dystrophic neurites, synaptic loss and neuronal death). Also in relation with the inflammatory response, at the end of 2014 we initiated a new collaborative project, funded by the Foundation La Marató of TV3, with several groups from CIBERNED, and coordinated by Dr. Elena Galea (University of Barcelona), on the properties of astrocytes and their role in the development of the AD pathology. On the other hand, this year we continued with the neuropathological characterization of transgenic PS1/APP models, focusing on the hippocampal subiculum region. This cerebral region is the first to be affected by the amyloid pathology and the neurodegenerative process including death of pyramidal neurons and interneurons. Results indicate that our PS1/APP model exhibits a degenerative phenotype similar to that described in patients in highly vulnerable cerebral regions, and thus represents a very valuable model for assessing the therapeutic capacity of neuroprotective strategies and reduce symptoms of the disease. Finally, we would like to highlight our translational research activity, and in this sense this year our group has conducted a project with the company Neuron Bio focused on preclinical study of potentials compounds with neuroprotective activity for AD. As a result of this collaboration and previous completed successfully, we have established a partnership with this company and other national groups for the development of first-in-class molecules as modifiers of the evolution of neurological/ degenerative diseases. Keywords Alzheimer, Neurodegeneration, Inflammation, Abeta, Hippocampus, Transgenic models, Neuropathology, Preclinical studies Publications Jimenez S, Navarro V, Moyano J, Sanchez-Mico M, Torres M, Davila JC, et al. Disruption of amyloid plaques integrity affects the soluble oligomers content from Alzheimer disease brains. PloS one. 2014;9(12):e Epub 2014 Dec 8. 65

72 Trujillo-Estrada L, Davila JC, Sanchez-Mejias E, Sanchez-Varo R, Gomez-Arboledas A, Vizuete M, et al. Early neuronal loss and axonal/presynaptic damage is associated with accelerated amyloid-β accumulation in AβPP/PS1 Alzheimer's disease mice subiculum. Journal of Alzheimer's disease: JAD. 2014;42(2): Cardona C, Sanchez-Mejias E, Davila JC, Martin-Rufian M, Campos-Sandoval JA, Vitorica J, et al. Expression of Gls and Gls2 glutaminase isoforms in astrocytes. Glia. 2015;63(3): Epub 2014 Oct 9. Galvan A, Hu X, Rommelfanger KS, Pare JF, Khan ZU, Smith Y, et al. Localization and function of dopamine receptors in the subthalamic nucleus of normal and parkinsonian monkeys. Journal of neurophysiology. 2014;112(2): Epub 2014 Apr 23. Khan ZU, Martin-Montanez E, Navarro-Lobato I, Muly EC. Memory deficits in aging and neurological diseases. Progress in molecular biology and translational science. 2014;122:1-29. Muly EC, Khan ZU. Preface. Molecular basis of memory. Progress in molecular biology and translational science. 2014;122:xv. Jimenez AJ, Rodriguez-Perez LM, Dominguez-Pinos MD, Gomez-Roldan MC, Garcia-Bonilla M, Ho- Plagaro A, et al. Increased levels of tumour necrosis factor alpha (TNFα) but not transforming growth factor-beta 1 (TGFβ1) are associated with the severity of congenital hydrocephalus in the hyh mouse. Neuropathology and applied neurobiology. 2014;40(7): Research Projects Code: 8.06/ Title: Evaluación in vivo de dos compuestos de interés. Utilización de estatinas neuroprotectoras y estrategias clínicas para enfermedades del sistema nervioso central Principal Investigator: Antonia Gutiérrez Pérez CIBERNED s groups: Type: CCAA Funding Agency: Neuron Biopharma Funding: Duration: 2014 Code: PI12/01431 Title: Oligómeros tóxicos del Abeta como agentes causantes de la disfunción del citoesqueleto y los procesos proteolíticos en la Principal Investigator: Antonia Gutiérrez Pérez CIBERNED s collaboration: Yes CIBERNED s groups: G411; G415 Type: International Funding Agency: Instituto de Salud Carlos III Funding: ,5 Duration: Code: PI2013/01 Title: Propiedades emergentes de la relación neurona-glía que subyacen a neurodegeneración y demencia en la enfermedad de Alzheimer Principal Investigator: Ignacio Torres-Alemán CIBERNED s collaboration: Yes CIBERNED s groups: G413; G204; G409; G108; G411; G415 Type: National Funding Agency: CIBERNED Funding: Duration: Code: CTS-1694 Title: Una estrategia para recuperar la memoria en el envejecimiento y la enfermedad de Alzheimer Principal Investigator: Zafaruddin Khan CIBERNED s groups: Type: CCAA Funding Agency: Junta de Andalucía. Proyectos de Excelencia Funding: Duration: 2014 Code: BFU R Title: Una estrategia para recuperar y prevenir la pérdida de la memoria Principal Investigator: Zafaruddin Khan CIBERNED s groups: Type: National Funding Agency: Ministerio de Economía y Competitividad Program 1 66

73 2014 Annual Report Antonia Gutiérrez Pérez Group Funding: Duration: Code: CTS 2035 Title: Oligomerización y toxicidad de los péptidos de Abeta: búsqueda de nuevas dianas de interes terapéutico en la enfermedad de Alzheimer Principal Investigator: Javier Vitorica CIBERNED s collaboration: Yes CIBERNED s groups: G415; G411 Type: CCAA Funding Agency: Junta de Andalucía Funding: Duration: Code: PI12/01439 Title: Oligómeros tóxicos del Abeta como agentes causantes de la disfunción del citoesqueleto y los procesos proteolíticos en la Principal Investigator: Javier Vitorica CIBERNED s collaboration: Yes CIBERNED s groups: G415; G411 Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration:

74 Group Alberto Lleó Bisa Program 1 CONTACT DETAILS oup Alberto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Lleó BisaGroup Unidad Alberto de Lleó Memoria Bisa Group Alberto ó Bisa Group Alberto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Lleó Bisa Servicio Group de Neurología Alberto Lleó Bisa Group Hospital de la Santa Creu i Sant Pau berto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Sant Antoni Lleó Bisa Mª Claret, Group 167 Alberto Lleó Bisa oup Alberto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Lleó BisaGroup Alberto Barcelona Lleó (Spain) BisaGroup Alberto Phone: ó Bisa Group Alberto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Lleó BisaGroup Fax: Alberto Lleó Bisa Group berto Lleó Bisa Group Alberto Lleó Bisa Group Alberto Lleó BisaGroup Alberto Lleó alleo@santpau.es BisaGroup Alberto Lleó Bisa LIST OF PERSONNEL Alberto Lleó Bisa Principal Investigator, Clinical Head, Neurology Rafael Blesa González Head of Department Jordi Clarimón Echavarria Postdoctoral Fellow Juan Fortea Ormaechea Researcher Staff Olivia Belvin Postdoctoral Fellow Oriol Dols Icardo PhD student Martí Colom Cadena PhD student Daniel Alcolea Rodríguez PhD student Isabel Sala Matavera PhD student Mª Belén Sánchez Saudinós PhD student María Carmona Iragui PhD student Sofía Antón Aguirre PhD student Laia Muñoz Llahuma Lab Technician Eduard Vilaplana Martínez PhD student Laura Cervera PhD student Marta Querol PhD student Estrella Morenas PhD student Roser Ribosa PhD student María Carmona-Iragui Río Hortega Researcher Victor Montal PhD student Isabel Sala PhD student Belén Sánchez-Saudinós PhD student 68

75 2014 Annual Report Group Alberto Lleó Bisa Summary Our group is composed of a multidisciplinary team of neurologists, neuropsychologists, biologists, engineers and lab technicians. In the last 5 years, the activities of the group have focused on translational and clinical aspects of neurodegenerative dementias. In particular, the group has been working on novel genetics factors and novel biomarkers of the most common neurodegenerative dementias. We have investigated new genetic risk variants in Alzheimer s disease (AD) and frontotemporal dementia (FTD). In parallel, the group has conducted different investigations in cerebrospinal fluid biomarkers along the continuum of AD, dementia with Lewy bodies, FTD and Down syndrome. The group is also involved in different imaging studies, using magnetic resonance imaging and amyloid positron emission tomography (PET) to determine the structural correlates of different biomarkers along the AD continuum, Down syndrome and other neurodegenerative dementias. Finally, the group also has uncovered important aspects of the molecular basis of neurodegenerative dementias by using cell culture models or human brain tissue. The group offers a unique environment for clinicians and investigators to tackle basic and translational aspects of neurodegeneration. Keywords Alzheimer, Dementia, Amyloid, Imaging Publications Podlesniy P, Figueiro-Silva J, Llado A, Antonell A, Sanchez-Valle R, Alcolea D, et al. Characterization of mtdna detection in cerebrospinal fluid. Reply: To PMID Annals of neurology. 2014;75(3): Epub 2014 Mar 7. Campanari ML, Garcia-Ayllon MS, Belbin O, Galceran J, Lleo A, Saez-Valero J. Acetylcholinesterase modulates presenilin-1 levels and γ-secretase activity. Journal of Alzheimer's disease: JAD. 2014;41(3): Gkampeta A, Fidani L, Clarimon J, Kalinderi K, Katopodi T, Zafeiriou D, et al. Association of brain-derived neurotrophic factor (BDNF) and elongator protein complex 4 (ELP4) polymorphisms with benign epilepsy with centrotemporal spikes in a Greek population. Epilepsy research. 2014;108(10): Epub 2014 Sep 22. Lleo A, Cavedo E, Parnetti L, Vanderstichele H, Herukka SK, Andreasen N, et al. Cerebrospinal fluid biomarkers in trials for Alzheimer and Parkinson diseases. Nature reviews. Neurology. 2015;11(1): Epub 2014 Dec 16. Fortea J, Vilaplana E, Alcolea D, Carmona-Iragui M, Sanchez-Saudinos MB, Sala I, et al. Cerebrospinal fluid β-amyloid and phospho-tau biomarker interactions affecting brain structure in preclinical Alzheimer disease. Annals of neurology. 2014;76(2): Epub 2014 Jun 13. Sanchez-Benavides G, Pena-Casanova J, Casals-Coll M, Gramunt N, Molinuevo JL, Gomez-Anson B, et al. Cognitive and neuroimaging profiles in mild cognitive impairment and Alzheimer's disease: data from the Spanish Multicenter Normative Studies (NEURONORMA Project). Journal of Alzheimer's disease: JAD. 2014;41(3): International Genomics of Alzheimer's Disease Consortium (IGAP), International Genomics of Alzheimer's Disease Consortium IGAP. Convergent genetic and expression data implicate immunity in Alzheimer's disease. Alzheimer's & dementia: the journal of the Alzheimer's Association. [Epub ahead of print]. Epub 2014 Dec

76 Compta Y, Valente T, Saura J, Segura B, Iranzo A, Serradell M, et al. Correlates of cerebrospinal fluid levels of oligomeric- and total-α-synuclein in premotor, motor and dementia stages of Parkinson's disease. Journal of neurology. 2014;262(2): Epub 2014 Nov 8. Manero RM, Casals-Coll M, Sanchez-Benavides G, Rodriguez-de los Reyes ON, Aguilar M, Badenes D, et al. Diagnostic validity of the Alzheimer's disease functional assessment and change scale in mild cognitive impairment and mild to moderate Alzheimer's disease. Dementia and geriatric cognitive disorders. 2014;37(5-6): Epub 2014 Feb 18. Suarez-Calvet M, Camacho V, Gomez-Anson B, Anton S, Vives-Gilabert Y, Dols-Icardo O, et al. Early Cerebellar Hypometabolism in Patients With Frontotemporal Dementia Carrying the C9orf72 Expansion. Alzheimer disease and associated disorders. [Epub ahead of print]. Epub 2014 Sep 8. Spataro N, Calafell F, Cervera-Carles L, Casals F, Pagonabarraga J, Pascual-Sedano B, et al. Mendelian genes for Parkinson's disease contribute to the sporadic forms of the disease. Human molecular genetics. [Epub ahead of print]. Epub 2014 Dec 11. Tajes M, Eraso-Pichot A, Rubio-Moscardo F, Guivernau B, Ramos-Fernandez E, Bosch-Morato M, et al. Methylglyoxal produced by amyloid-β peptide-induced nitrotyrosination of triosephosphate isomerase triggers neuronal death in Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;41(1): Querol L, Nogales-Gadea G, Rojas-Garcia R, Diaz-Manera J, Pardo J, Ortega-Moreno A, et al. Neurofascin IgG4 antibodies in CIDP associate with disabling tremor and poor response to IVIg. Neurology. 2014;82(10): Epub 2014 Feb 12. De la Torre GG, Suarez-Llorens A, Caballero FJ, Ramallo MA, Randolph C, Lleo A, et al. Norms and reliability for the Spanish version of the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) Form A. Journal of clinical and experimental neuropsychology. 2014;36(10): Epub 2014 Nov 3. Ramos-Fernandez E, Tajes M, Palomer E, Ill-Raga G, Bosch-Morato M, Guivernau B, et al. Posttranslational nitro-glycative modifications of albumin in Alzheimer's disease: implications in cytotoxicity and amyloid-β peptide aggregation. Journal of Alzheimer's disease: JAD. 2014;40(3): Garcia-Ayllon MS, Campanari ML, Montenegro MF, Cuchillo-Ibanez I, Belbin O, Lleo A, et al. Presenilin-1 influences processing of the acetylcholinesterase membrane anchor PRiMA. Neurobiology of aging. 2014;35(7): Epub 2014 Feb 6. Alcolea D, Carmona-Iragui M, Suarez-Calvet M, Sanchez-Saudinos MB, Sala I, Anton-Aguirre S, et al. Relationship between β-secretase, inflammation and core cerebrospinal fluid biomarkers for Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;42(1): Medway C, Combarros O, Cortina-Borja M, Butler HT, Ibrahim-Verbaas CA, de Bruijn RF, et al. The sexspecific associations of the aromatase gene with Alzheimer's disease and its interaction with IL10 in the Epistasis Project. European journal of human genetics: EJHG. 2014;22(2): Epub 2013 Jun 5. Bocchetta M, Galluzzi S, Kehoe PG, Aguera E, Bernabei R, Bullock R, et al. The use of biomarkers for the etiologic diagnosis of MCI in Europe: An EADC survey. Alzheimer's & dementia: the journal of the Alzheimer's Association. [Epub ahead of print]. Epub 2014 Aug 20. Olde Rikkert MG, Verhey FR, Blesa R, von Arnim CA, Bongers A, Harrison J, et al. Tolerability and Safety of Souvenaid in Patients with Mild Alzheimer's Disease: Results of Multi-Center, 24-Week, Open-Label Extension Study. Journal of Alzheimer's disease: JAD. 2015;44(2): Epub 2014 Oct 16. Ferrari R, Hernandez DG, Nalls MA, Rohrer JD, Ramasamy A, Kwok JB, et al. Frontotemporal dementia and its subtypes: a genome-wide association study. Lancet neurology. 2014;13(7): Escott-Price V, Bellenguez C, Wang LS, Choi SH, Harold D, Jones L, et al. Gene-wide analysis detects two new susceptibility genes for Alzheimer's disease. PloS one. 2014;9(6):e Epub 2014 Jun 12. Thelen M, Razquin C, Hernandez I, Gorostidi A, Sanchez-Valle R, Ortega-Cubero S, et al. Investigation of the role of rare TREM2 variants in frontotemporal dementia subtypes. Neurobiology of aging. 2014;35(11):2657.e13-9. Epub 2014 Jun 20. van der Zee J, Van Langenhove T, Kovacs GG, Dillen L, Deschamps W, Engelborghs S, et al. Rare mutations in SQSTM1 modify susceptibility to frontotemporal lobar degeneration. Acta neuropathologica. 2014;128(3): Epub 2014 Jun 5. Program 1 70

77 2014 Annual Report Group Alberto Lleó Bisa Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agundez JA, Jimenez-Jimenez FJ, Ross OA, et al. TREM2 R47H variant and risk of essential tremor: A cross-sectional international multicenter study. Parkinsonism & related disorders. [Epub ahead of print]. Epub 2014 Dec 24. Dols-Icardo O,Garcia-Redondo A, Rojas-Garcia R, Sanchez-Valle R, Noguera A, Gomez-Tortosa E, et al. Characterization of the repeat expansion size in C9orf72 in amyotrophic lateral sclerosis and frontotemporal dementia. Human molecular genetics. 2014;23(3): Casals-Coll M, Sanchez-Benavides G, Meza-Cavazos S, Manero RM, Aguilar M, Badenes D, et al. Spanish multicenter normative studies (NEURONORMA project): normative data and equivalence of four BNT short-form versions. Archives of clinical neuropsychology: the official journal of the National Academy of Neuropsychologists. 2014;29(1): Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodriguez-Rodriguez E, Lopez de Munain A, et al. Assessing the role of the TREM2 p.r47h variant as a risk factor for Alzheimer's disease and frontotemporal dementia. Neurobiology of aging. 2014;35(2):444.e1-4. Marti-Fàbregas J, Delgado-Mederos R, Marin R, de la Ossa NP, Alonso de Lecinana M, Rodriguez- Yanez M, et al. Prognostic value of plasma β-amyloid levels in patients with acute intracerebral hemorrhage. Stroke: a journal of cerebral circulation. 2014;45(2): Suarez-Calvet M, Belbin O, Pera M, Badiola N, Magrane J, Guardia-Laguarta C, et al. Autosomaldominant Alzheimer's disease mutations at the same codon of amyloid precursor protein differentially alter Aβ production. Journal of neurochemistry. 2014;128(2): Suarez-Calvet M, Dols-Icardo O, Llado A, Sanchez-Valle R, Hernandez I, Amer G, et al. Plasma phosphorylated TDP-43 levels are elevated in patients with frontotemporal dementia carrying a C9orf72 repeat expansion or a GRN mutation. Journal of neurology, neurosurgery, and psychiatry. 2014;85(6): Alcolea D, Martinez-Lage P, Izagirre A, Clerigue M, Carmona-Iragui M, Alvarez RM, et al. Feasibility of lumbar puncture in the study of cerebrospinal fluid biomarkers for Alzheimer's disease: a multicenter study in Spain. Journal of Alzheimer's disease: JAD. 2014;39(4): Research Projects Code: PI11/02425 Title: Estudios multimodales de líquido cefalorraquídeo y resonancia magnética en la enfermedad de Alzheimer preclínica Principal Investigator: Juan Fortea CIBERNED s groups: Type: International Funding Agency: ISCIII Funding: N.D. Duration: Code: 2013/07A Title: Papel de GSK-3 β en las alteraciones de los circuitos corticales que ocurren en la enfermedad de Alzheimer Principal Investigator: Teresa Iglesias Vacas CIBERNED s collaboration: Yes CIBERNED s groups: G401; G403; G111; G504; G307 Type: International Funding Agency: CIBERNED Funding: Duration: Code: PI12/01311 Title: Variantes genéticas raras y su implicación en la enfermedad de Alzheimer: uso de nuevas tecnologías de ultrasecuenciación para el estudio de genes implicados en la enfermedad Principal Investigator: Jordi Clarimon CIBERNED s groups: Type: International Funding Agency: ISCIII Funding: N.D. Duration: Code: PI11/ Title: Biomarkers for Alzheimer s disease and Parkinson s disease (BIOMARKAPD) 71

78 Principal Investigator: Javier Sáez Valero CIBERNED s collaboration: Yes CIBERNED s groups: G504; G407 Type: European Funding Agency: ISCIII Funding: Duration: Code: PI13/01532 Title: Enfermedad de Alzheimer y síndrome de Down. Estudios multimodales de líquido cefalorraquídeo, resonancia magnética y PET de amiloide Principal Investigator: Rafael Blesa CIBERNED s groups: Type: International Funding Agency: Fondo de Investigación Sanitario. ISCIII Funding: Duration: Program 1 72

79 Group María López de Ceballos Lafarga Program 1 CONTACT DETAILS Group María López de Ceballos Lafarga Group María López de Ceballos Lafarga Group María Instituto López Cajal, de Ceballos CSIC Lafarga Group María López de Ceballos Lafarga Group María López de Ceballos Lafarga Group María Avenida López Doctor de Ceballos Arce, Lafarga 37 Group Mar Madrid (Spain) López de Ceballos Lafarga Group María López de Ceballos Lafarga Group María López Phone: de Ceballos Lafarga 854 Group 716 María López d Fax: Ceballos Lafarga Group María López de Ceballos Lafarga Group María López de Ceballos Lafarga Group María López de Ceballo mceballos@cajal.csic.es Lafarga Group María López de Ceballos Lafarga Group María López de Ceballos Lafarga Group María López de Ceballos Lafarg LIST OF PERSONNEL María López de Ceballos Lafarga Principal Investigator, Tenure Scientist, CSIC Silvia de Vidania Ballesteros PhD student Zayra Elisa Hernández Núñez Student Nathaniel Wilson Student Nora Kadiri Moreno Student Sahba Seddhigi Student Clara González Martínez Student 73

80 Summary Cannabinoids exert different beneficial effects on several neurodegenerative diseases, including Alzheimer s disease (AD). Furthermore, CB2 receptor agonists, which lack psychoactive effects, may be interesting for their translation into the Clinics. In a previous work (Martín-Moreno et al., J Neuroinflamm. 2012;9:8) we have shown that prolonged oral administration of the CB2 selective agonist JWH-133 diminished microglial activation, the enhanced inflammatory parameters and β-amyloid levels, and normalized the cognitive deficits of Tg APP mice, an AD model. In addition, such treatment reversed the decreased in 18F-deoxyglucose uptake in hippocampus and cortical regions assessed by PET. The brain relies upon glucose as its primary fuel, and increased glucose supply facilitates cognitive functions. Therefore we have studied the effects of CB2 receptor activation on glucose uptake. The additionof both selective as well as non-selective CB2R agonists rapidly stimulate glucose uptake in vitro and in vivo, in a manner that is sensitive to the selective CB2R antagonist, AM630. Glucose uptake is enhanced: a) in cultures of neocortical astrocytes and neurons, b) in acute hippocampal slices, c) in different brain areas of young adult male C57Bl/6j or CD-1 mice and Wistar rats, as well as in middle-aged C57Bl/6j mice. Inhibiting cyclooxygenase-2 (COX-2) with DuP697, an enzyme that metabolizes the endogenous CB2R agonist anandamide in brain, also stimulates the uptake of glucose in hippocampal slices of middle-aged mice, an affect that was again prevented by AM630. However, this was not the case in middle-aged TgAPP 2576 mice where DuP697 had no such effect, consistent with the reduction in anandamide that we found in the hippocampus of these mice, which explains why COX-2 inhibition failed to stimulate glucose uptake via CB2Rs. An inhibitor of fatty acid amidohydrolase (FAAH) lacked effect, likely because the expression of that enzyme was unaffected in Tg APP mice. Together, these results reveal a novel general glucoregulatory role for CB2Rs in the brain, raising therapeutic interest in CB2R agonists as nootropic agents. On the other hand, we have recently described the similarities and differences between FAAH activity and its modulation in normal and pathological ageing. FAAH activity was analysed in an independent cohort of human cortical membrane samples from control and AD patients, and in membrane and synaptosomes from adult and aged (28 months) rat cerebral cortex. Our results demonstrate that FAAH activity: (a) Decreases in frontal cortex from human patients with AD and this effect is mimicked by Aβ1-40 peptide; (b) Increases and decreases in aged rat cerebrocortical membrane and synaptosomes, respectively; (c) In the presence of JWH-133 (a CB2 selective agonist), c.i) increases slightly in human controls; c.ii) decreases in adults and it is completely abolished in aged rat cerebrocortical membrane; and c.iii) decreases in adult and aged rat cerebrocortical synaptosomes; (d) In the presence of WIN 55,212-2 (a mixed CB1/ CB2 agonist); d.i) decreases in human controls and increases in AD patients; and d.ii) decreases in adult and aged rat cerebrocortical membrane and synaptosomes. Although a similar profile is observed in fatty acid amide hydrolase activity between aged rat synaptic endings and human Alzheimer s disease brains, it is differently modulated by CB1/CB2 agonists. This modulation leads to a reduced availability of anandamide in AD and to an increased availability of this endocannabinoid in ageing. Keywords Alzheimer s disease, β-amyloid, Cannabinoids, Cyclooxygenase-2, Fatty acid amydohydrolase, Glucoregulation, Glucose uptake Program 1 74

81 2014 Annual Report María López de Ceballos Lafarga Group Publications Pascual AC, Martin-Moreno AM, Giusto NM, de Ceballos ML, Pasquare SJ. Normal aging in rats and pathological aging in human Alzheimer's disease decrease FAAH activity: Modulation by cannabinoid agonists. Experimental gerontology. 2014;60:92-9. Epub 2014 Oct 18. Perez-Gonzalez R, Alvira-Botero MX, Robayo O, Antequera D, Garzon M, Martin-Moreno AM, et al. Leptin gene therapy attenuates neuronal damages evoked by amyloid-β and rescues memory deficits in APP/PS1 mice. Gene therapy. 2014;21(3): Research Projects Code: S2010/BMD-2349 Title: Imagen Molecular Multimodal de la Inflamación Principal Investigator: S Cerdan CIBERNED s groups: Type: CCAA Funding Agency: CAM Funding: Duration: Code: BFU C03-01 Title: Señalización de NOTCH/NGN3 en las acciones neuroprotectoras de compuestos Principal Investigator: Luis Miguel García Segura CIBERNED s groups: Type: National Funding Agency: MCINN Funding: Duration:

82 Group Carlos Matute Almau Program 1 CONTACT DETAILS oup Carlos Matute Almau Group Carlos Matute Almau Group Carlos Matute Department Almau of Group Neurosciences Carlos Matute Almau atute Almau Group Carlos Matute Almau Group Carlos Matute Almau Facultad Group de Medicina Carlos Matute y Odontología Almau Group Carlos Universidad del País Vasco rlos Matute Almau Group Carlos Matute Almau Group Carlos Matute Almau Group Carlos Leioa Matute (Spain) Almau Group oup Carlos Matute Almau Group Carlos Matute Almau Group Carlos Matute Phone: Almau +34 Group Carlos 244 Matute Almau Fax: atute Almau Group Carlos Matute Almau Group Carlos Matute Almau Group Carlos carlos.matute@ehu.es Matute Almau Group Carlos rlos Matute Almau Group Carlos Matute Almau Group Carlos Matute Almau Group Carlos Matute Almau Group LIST OF PERSONNEL Carlos Matute Almau Principal Investigator, Full Professor Alberto Pérez Samartín Associate Professor Fernando Pérez Cerdá Associate Professor Elena Alberdi Alfonso Assistant Professor María Domercq García Assistant Professor María Victoria Sánchez Gómez Assistant Professor Susana Mato Santos Ramón y Cajal Researcher Fabio Cavaliere Research Associate Asier Ruiz Núñez Postdoctoral Fellow Abraham Cisneros Mejorado Postdoctoral Fellow Ane Wissenbach PhD student Tania Quintela PhD student Andrea Manterola Juaristi PhD student Hazel Gómez Manrique Technician CIBERNED Saioa Marcos Ezquerro Technician CIBERNED 76

83 2014 Annual Report Carlos Matute Almau Group Summary The main goal of our laboratory is to unveil the pathological mechanisms of glial cells in Alzheimer disease (AD) and their contribution to disease progression. Recently, we have described novel mechanism by which the toxic soluble amyloid beta (Aβ1-42) peptide modulates integrin β1 activity and triggers astrogliosis by reactive oxygen species (ROS) generation through NOX-2 activation in vitro and in vivo. In this year, we have defined that these markers of oxidative and astroglial stress as well as integrin β1 are upregulated in Aβ1-42-treated astrocytes in culture, hippocampus of triple transgenic mice that models EA, and in postmortem brains of individuals with AD (Braak V and VI/C). Moreover, overexpression of integrin β1, NOX2 and GFAP levels correlates with amyloid burden in hippocampus of 18 month-old triple transgenic mice, which develop a dramatic acumulation of Aβ1-42. Immunohistochemical analysis showed that integrin β1 and NOX-2 levels were significantly higher in reactive astrocytes in triple-transgenic mice than in wild type mice, as well as in AD brains as compared to controls. These data suggest that Aβ1-42 may directly cause and exacerbate astrocyte reactivity in AD by enhancing integrin β1 and NOX activity via ROS-dependent mechanisms. On the other hand, we have observed that Aβ1-42 has a dual effect on the oligodendroglial lineage as it upregulates the levels of myelin proteins (MBP, CNPase and PLP) and increases PDGFα receptor expression in progenitors. In the last year, we have found that Aβ1-42 causes a sustained Src and CREB protein phosphorylation in primary cultures of oligodendrocytes as well as in organotypic cerebellar slices, and that specific pharmacological inhibition of both pathways reduces the Aβ-induced MBP upregulation. In addition, MBP levels in hippocampus and corpus callosum of 18-month old triple transgenic mice are increase in both regions and correlate with Aβ oligomer burden. Consistent with those findings, MBP levels in frontal cortex of AD were significantly higher than in brains of non-demented controls. Together, these date support a role of Aβ1-42 and Src-like kinases in the pathophysiology of myelin in AD. Keywords β-amyloid toxicity, Astrocytes, Oligodendrocytes, Myelin, Oxidative stress Publications Moreno B, Lopez I, Fernandez-Diez B, Gottlieb M, Matute C, Sanchez-Gomez MV, et al. Differential neuroprotective effects of 5'-deoxy-5'-methylthioadenosine. PloS one. 2014;9(3):e Epub 2014 Mar 5. Cisneros-Mejorado A, Perez-Samartin A, Gottlieb M, Matute C. ATP signaling in brain: release, excitotoxicity and potential therapeutic targets. Cellular and molecular neurobiology. 2015;35(1):1-6. Epub 2014 Aug 6. Bernal-Chico A, Canedo M, Manterola A, Sanchez-Gomez MV, Perez-Samartin A, Rodriguez-Puertas R, et al. Blockade of monoacylglycerol lipase inhibits oligodendrocyte excitotoxicity and prevents demyelination in vivo. Glia. 2015;63(1): Epub 2014 Aug 8. Alberdi E, Ruiz A, Matute C. Calcium dyshomeostasis in white matter injury. Springer Series in Translational Stroke Research Epub Ruiz A, Alberdi E, Matute C. CGP37157, an inhibitor of the mitochondrial Na+/Ca2+ exchanger, protects neurons from excitotoxicity by blocking voltage-gated Ca2+ channels. Cell death & disease. 2014;5:e1156. Epub 2014 Apr

84 Matute C, Stys PK. Editors' preface: the colourful white matter. Glia. 2014;62(11): Soria FN, Perez-Samartin A, Martin A, Gona KB, Llop J, Szczupak B, et al. Extrasynaptic glutamate release through cystine/glutamate antiporter contributes to ischemic damage. The Journal of clinical investigation. 2014;124(8): Epub 2014 Jul 18. Cipriani R, Domercq M, Matute C. Ischemia and stroke. In: Tremblay ME, Sierra A, eds. Microglia in health and disease Epub Butt AM, Fern RF, Matute C. Neurotransmitter signaling in white matter. Glia. 2014;62(11): Epub 2014 Apr 21. Gutierrez-Fernandez A, Palomino A, Gonzalez-Pinto A, Ugarte A, Hernanz M, Mendibil B, et al. Novel association of Neuregulin 1 gene with bipolar disorder but not with schizophrenia. Schizophrenia research. 2014;159(2-3): Epub 2014 Sep 20. Sanchez-Gomez MV, Perez-Cerda F, Matute C. White matter damage in multiple sclerosis. Springer Series in Translational Stroke Research Epub Baltan S, Carmichael ST, Matute C, Xi G, Zhang JH. White matter injury in stroke and in CNS diseases. Springer Series in Translational Stroke Research Epub Fern RF, Matute C, Stys PK. White matter injury: Ischemic and nonischemic. Glia. 2014;62(11): Epub 2014 Jul 18. Vazquez-Villoldo N, Domercq M, Martin A, Llop J, Gomez-Vallejo V, Matute C. P2X4 receptors control the fate and survival of activated microglia. Glia. 2014;62(2): Gonzalez-Fernandez E, Sanchez-Gomez MV, Perez-Samartin A, Arellano RO, Matute C. A3 Adenosine receptors mediate oligodendrocyte death and ischemic damage to optic nerve. Glia. 2014;62(2): Research Projects Code: Title: Assessment of the neuroprotective activity of FTY720 (fingolimod) in experimental models of brain damage Principal Investigator: Carlos Matute Almau CIBERNED s groups: Type: International Funding Agency: Novartis Funding: Duration: Code: IT Title: Bases moleculares y celulares de la neurodegeneración Principal Investigator: Carlos Matute Almau CIBERNED s groups: Type: CCAA Funding Agency: Gobierno Vasco Funding: Duration: Code: Merck KGaA Title: Could reeducating microglia by P2X4R manipulation constitute an alternative therapy for MS? Principal Investigator: María Domercq García CIBERNED s groups: Type: International Funding Agency: Merck KGaA Funding: Duration: Code: PRI-PIMNEU Title: Nanostroke Principal Investigator: Carlos Matute Almau CIBERNED s groups: Type: International Funding Agency: ERANET Neuron Funding: Duration: Code: Title: Neurogenic potential of dimethyl-fumarate Principal Investigator: Carlos Matute Almau CIBERNED s groups: Program 1 78

85 2014 Annual Report Carlos Matute Almau Group Type: International Funding Agency: Biogen Pharma Funding: Duration: Code: ARSEP2013 Title: Therapeutic potential of monoacylglycerol lipase inhibition for the treatment of multiple sclerosis Principal Investigator: Carlos Matute Almau CIBERNED s groups: Type: International Funding Agency: ARSEP Foundation (France) Funding: Duration: Code: 2014/06 Title: Inicio y Progresión de la Enfermedad de Parkinson: Papel de la Activacion Glial Principal Investigator: María Cruz Rodríguez Oroz CIBERNED s collaboration: Yes CIBERNED s groups: G206; G404; G205 Type: International Funding Agency: N.D. Funding: Duration: PhD Dissertations Author: Ana Bernal Chico Title: Modulation of brain endocannabinoid system and therapeutic potential of monoacylglycerol lipase blockade during demyelination in vivo Date: 22nd July 2014 Supervisor: Susana Mato Santos 79

86 Group Guadalupe Mengod de los Arcos Program 1 CONTACT DETAILS oup Guadalupe Mengod de los Arcos Group Guadalupe Mengod de los Arcos IIBB-CSIC-IDIBAPS-CIBERNED Group Guadalupe Mengod de los Arcos oup Guadalupe Mengod de los Arcos Group Guadalupe Mengod de los Arcos Phone: Group Guadalupe Mengod de los cos Group Guadalupe Mengod de los Arcos Group Guadalupe Mengod de los Arcos Fax: +34 Group Guadalupe 301 Mengod de guadalupe.mengod@iibb.csic.es s Arcos Group Guadalupe Mengod de los Arcos Group Guadalupe Mengod de los Arcos Group Guadalupe Mengod LIST OF PERSONNEL Guadalupe Mengod Principal Investigator, Full Professor, CSIC Roser Cortés Tenure Scientist, CSIC Mª Teresa Vilaró Tenure Scientist, CSIC Giuseppe Mocci PhD student Núria Paúl-Fernández PhD student Rocío Martín-Álvarez Lab Technician CIBERNED Silvia Serrano Advanced Technician CIBERNED Giovanni Di Pinto Erasmus Researcher Samantha Sperduti PhD student Daniel Castro Engstrom Masters Researcher Estela Núñez Manchón Masters Researcher Marina De San Agustín Márquez Student Belén Gago Calderón Postdoctoral Fellow 80

87 2014 Annual Report Guadalupe Mengod de los Arcos Group Summary Our group is composed of three research sublines, which have the main and common research theme of neurodegeneration. In one of the sublines, we are studying the expression of neuronal markers in dementia, argyrophilic grain disease and Alzheimer s disease by histochemical methods and RT-PCR, and analyzing the differential expression of proteins in these two diseases by proteomics. We identified several proteins that are differentially expressed in patients with argyrophilic grain disease, Alzheimer s disease and control cases by proteomic analysis of brain samples. We are now validating these alterations. In particular we have analyzed the expression of protein zeta in relation to the presence of neurofibrillary tangles, and annexin A5 in the hippocampus and entorhinal and perirhinal cortices. We observed that annexin A5 is increased in Alzheimer s cases and, to a lesser extent, in argyrophilic grain disease, in association with astrocytes, blood vessels and amyloid plaques. Part of this study has been the research work of the Master in Biomedicine of the UB presented by Estela Nuñez. On the other hand, we investigated the activation of 5-HT4 serotonin receptors as an approach to interfere with Aß amyloid peptide deposition in the 3xTg-AD mouse model of Alzheimer s disease. We observed that continued stimulation of these receptors resulted in very marked improvements in learning and memory, in parallel with a decrease of both intraneuronal Aß and extracellular amyloid load. The treatment attenuated neophobia and anxiety and increased cell proliferation in the two neurogenic areas of the adult brain. We have also determined that 5-HT4 receptors present in forebrain cholinergic regions of the rat brain are expressed in GABAergic and glutamatergic neurons present in those regions but were not detected in cholinergic neurons (Peñas-Cazorla R et al., 2014 [Epub ahead of print]). These results indicate that the known increase in acetylcholine release mediated by stimulation of 5-HT4 receptors does not occur through a direct action on cholinergic neurons as had been postulated in the literature, but rather through an action on other neuronal types that do express these receptors. And in the third subline of work, neuroinflammation, we are analyzing the molecular regulation of camp signaling in the murine experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis and looking for innovative drugs for multiple sclerosis treatment through the selective inhibition of camp-specific phosphodiesterases. Chronic treatment with the specific inhibitors roflumilast (PDE4) and C33 (PDE4B) of EAE mice produced a significant improvement in clinical symptoms in animals. We studied the effect of the treatment on lymphocyte infiltration, axonal damage and demyelination in the spinal cord as well as microglial and astrocytic reactivity. This work constituted the doctoral thesis of Paul Nuria Fernandez directed by the PI. We have also studied the effect of treatment with PDE7 inhibitors in EAE mice on the above parameters. The results are part of a manuscript submitted to publish and is under review. We analyzed the effects of stimulation of cultured microglia cell line Bv-2 and its treatment with the various PDE inhibitors on the polarization toward M1 (proinflammatory) or M2 (anti-inflammatory) phenotypes. We have observed, for example, that treatment with rolipram (PDE4 inhibitor) of Bv-2 cells stimulated with LPS promotes a shift of the cells towards an M2 spectrum population. This has been part of a Master in Translational Medicine, UB, presented by Daniel Castro. Keywords Neurodegeneration, Neuroinflammation, Argyrophilic gran disease, Human brain, Molecular Neuropharmacology 81

88 Publications Penas-Cazorla R, Vilaro MT. Serotonin 5-HT4 receptors and forebrain cholinergic system: receptor expression in identified cell populations. Brain structure & function. [Epub ahead of print]. Epub 2014 Sep 3. Marin C, Bonastre M, Mengod G, Cortes R, Giralt A, Obeso JA, et al. Early L-dopa, but not pramipexole, restores basal ganglia activity in partially 6-OHDA-lesioned rats. Neurobiology of disease. 2014;64: Mocci G, Jimenez-Sanchez L, Adell A, Cortes R, Artigas F. Expression of 5-HT2A receptors in prefrontal cortex pyramidal neurons projecting to nucleus accumbens. Potential relevance for atypical antipsychotic action. Neuropharmacology. 2014;79: Research Projects Code: PI10/01874 Title: El papel de las fosfodiesterasas del AMPc en la evolución y tratamiento de la esclerosis múltiple en el modelo de EAE en ratón. Nuevas aproximaciones terapéuticas Principal Investigator: Guadalupe Mengod CIBERNED s groups: Type: National Funding Agency: Instituto de Salud Carlos III, FIS Funding: Duration: Code: 2013/R/25 Title: Relationship between cholinergic dysfuntion and inflammation: study in multiple sclerosis; studies in EAE mice and RRMS patients Principal Investigator: Ada María Tata CIBERNED s groups: Type: International Funding Agency: Fondazione Italiana Sclerosis Multipla (FISM) Funding: Duration: PhD Dissertations Author: Giuseppe Mocci Title: Dianas de fármacos antipsicóticos en neuronas de proyección de la corteza prefrontal Date: 7th February 2014 Supervisor: Roser Cortés Colomé Program 1 82

89 Group María Asunción Pastor Muñoz Program 1 CONTACT DETAILS Group María Asunción Pastor Muñoz Group María Asunción Pastor Center Muñoz for Group Applied María Medical Asunción Research Pastor Muñoz Group María Asunción Pastor Muñoz Group María Asunción Pastor Muñoz Group María Universidad Asunción Pastor de Navarra Muñoz Group Mar Asunción Pastor Muñoz Group María Asunción Pastor Muñoz Group María Asunción Pastor Pamplona Muñoz (Spain) Group María Asunció Phone: Pastor Muñoz Group María Asunción Pastor Muñoz Group María Asunción Pastor Muñoz Group María Asunción Pasto Fax: Muñoz Group María Asunción Pastor Muñoz Group María Asunción Pastor Muñoz Group mapastor@unav.es María Asunción Pastor Muño Group María Asunción Pastor Muñoz Group María Asunción Pastor Muñoz Group María Asunción Pastor Muñoz Group LIST OF PERSONNEL María Asunción Pastor Muñoz Principal Investigator Pau Pastor Muñoz Co-Principal Investigator María Asunción Fernández Seara PhD Elisa Mengual Poza PhD Elkin Luis García PhD student Martin Martínez Villar PhD student Gabriel García Castellanos PhD student Luis Eudave Ramos PhD student Silvia Zarraluqui López PhD student Sara Ortega Cubero PhD student Gonzalo Arrondo PhD student Miquel Aguilar Barberá PhD Laura Molina Porcel PhD Juan Pablo Tartari Specialist Silvia Romero Contreras Nurse 83

90 Summary The research of Neuroimaging/Neurogenetics group at the CIMA is focused in the study of genetic markers of neurodegenerative diseases such as dementia or parkinsonisms either familial or sporadic. We are currently performing last generation sequencing of the human exome and transcriptome to identify novel genes involved in such disorders. As a result of this work, and from national and international collaborations in the genomics core, we have published 13 articles with many groups of CIBERNED, most of them in peer-reviewed International journals located at the first 25% of the IF ranking. Most publications report new knowledge on the genetic risk linked to different kinds of dementia and parkinsonism. In three of the article we reported the association of certain genetic variants with specific neuroimaging degenerative patterns. In fact, as an example of our activity, we identified frototemporal regions specially impaired among TREM2 R47H variant carriers, as well as the accurate characterization of loss of substantia nigra neural cells among subjects with sporadic and monogenic Parkinson disease, that can be used as a diagnostic marker. With regards to most modern neuroimaging sequences, we have updated with success perfusion MRI Arterial Spin Labeling (ASL) and BOLD sequences. The result of this work resulted on four articles detailing the optimization of an ASL MRI protocol in the mark of EU COST program. Many of the publication are fruit of a close collaboration with international groups and CIBERNED Spanish groups in the mark of DEGESCO Consortium, which groups many leader national groups expert in neurogenetics. The DEGESCO Consortium has already published two articles and one more is under review. The lines of our group will continue focusing on neuroimaging and genetic markers with diagnostic practical utility that will allow to design more specific clinical trials and hopefully new effective therapies. Keywords Neuroimaging, Functional neuroimaging, Genetics, SNPs, Gene Publications International Genomics of Alzheimer's Disease Consortium (IGAP), International Genomics of Alzheimer's Disease Consortium IGAP. Convergent genetic and expression data implicate immunity in Alzheimer's disease. Alzheimer's & dementia: the journal of the Alzheimer's Association. [Epub ahead of print]. Epub 2014 Dec 20. Zamurs LK, Idoate MA, Hanssen E, Gomez-Iba Ntildeez A, Pastor P, Lamand Eacute SR. Aberrant Mitochondria in a Bethlem Myopathy Patient with a Homozygous Amino Acid Substitution that Destabilises the Collagen VI α2(vi) chain. The Journal of biological chemistry. [Epub ahead of print]. Epub 2014 Dec 22. Lorenzo-Betancor O, Ogaki K, Soto-Ortolaza A, Labbe C, Vilarino-Güell C, Rajput A, et al. Analysis of nuclear export sequence regions of FUS-Related RNA-binding proteins in essential tremor. Plos One. 2014;9(11):e Epub 2014 Nov 6. Program 1 84

91 2014 Annual Report María Asunción Pastor Muñoz Group Ortega-Cubero S, Pagola I, Luquin MR, Viteri C, Pastor P, Gallego Perez-Larraya J, et al. Clinical and neuroimaging characteristics of 14 patients with prionopathy: A descriptive study. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Feb 25. Marta Vidorreta, Evelyne Balteau, Ze Wang, Enrico De Vita, Maria A. Pastor, David L. Thomas, et al. Evaluation of Segmented 3D Acquisition Schemes for Whole-brain High-resolution Arterial Spin Labeling at 3T. NMR Biomed. 2014;27(11): Cancela J, Pastorino M, Arredondo MT, Nikita KS, Villagra F, Pastor MA. Feasibility study of a wearable system based on a wireless body area network for gait assessment in Parkinson's disease patients. Sensors. 2014;14: Luis EO, Ortega-Cubero S, Lamet I, Razquin C, Cruchaga C, Benitez BA, et al. Frontobasal gray matter loss is associated with the TREM2 p.r47h variant. Neurobiology of aging. 2014;35(12): Epub 2014 Jun 17. Ferrari R, Hernandez DG, Nalls MA, Rohrer JD, Ramasamy A, Kwok JB, et al. Frontotemporal dementia and its subtypes: a genome-wide association study. Lancet neurology. 2014;13(7): Bras J, Guerreiro R, Darwent L, Parkkinen L, Ansorge O, Escott-Price V, et al. Genetic analysis implicates APOE, SNCA and suggests lysosomal dysfunction in the etiology of dementia with Lewy bodies. Human molecular genetics. 2014;23(23): Epub 2014 Jun 27. Escott-Price V, Bellenguez C, Wang LS, Choi SH, Harold D, Jones L, et al. Gene-wide analysis detects two new susceptibility genes for Alzheimer's disease. PloS one. 2014;9(6):e Epub 2014 Jun 12. Theuns J, Verstraeten A, Sleegers K, Wauters E, Gijselinck I, Smolders S, et al. Global investigation and meta-analysis of the C9orf72 (G4C2)n repeat in Parkinson disease. Neurology. 2014;83(21): Epub 2014 Oct 17. Thelen M, Razquin C, Hernandez I, Gorostidi A, Sanchez-Valle R, Ortega-Cubero S, et al. Investigation of the role of rare TREM2 variants in frontotemporal dementia subtypes. Neurobiology of aging. 2014;35(11):2657.e13-9. Epub 2014 Jun 20. Martinez M, Villagra F, Loayza F, Vidorreta M, Arrondo G, Louis E, et al. MRI-compatible Device for Examining Brain Activation Related to Stepping. IEEE Transactions on Medical Imaging. 2014;33(5): Sharma M, Wenning G, Krüger R;European Multiple-System Atrophy Study Group (EMSA-SG). Mutant COQ2 in multiple-system atrophy. The New England journal of medicine. 2014;371(1):80-1. Fachal L, Mosquera-Miguel A, Pastor P, Ortega-Cubero S, Lorenzo E, Oterino-Duran A, et al. No evidence of association between common European mitochondrial DNA variants in Alzheimer, Parkinson, and migraine in the Spanish population. American journal of medical genetics Part B Neuropsychiatric genetics: the official publication of the International Society of Psychiatric Genetics. 2015;168(1): Epub 2014 Oct 28. van der Zee J, Van Langenhove T, Kovacs GG, Dillen L, Deschamps W, Engelborghs S, et al. Rare mutations in SQSTM1 modify susceptibility to frontotemporal lobar degeneration. Acta neuropathologica. 2014;128(3): Epub 2014 Jun 5. Bernacer J, Balderas G, Martinez-Valbuena I, Pastor MA, Murillo JI. The problem of consciousness in habitual decision making. The Behavioral and brain sciences. 2014;37(1):21-2. Epub 2014 Jan 24. Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agundez JA, Jimenez-Jimenez FJ, Ross OA, et al. TREM2 R47H variant and risk of essential tremor: A cross-sectional international multicenter study. Parkinsonism & related disorders. [Epub ahead of print]. Epub 2014 Dec 24. Dols-Icardo O, Garcia-Redondo A, Rojas-Garcia R, Sanchez-Valle R, Noguera A, Gomez-Tortosa E, et al. Characterization of the repeat expansion size in C9orf72 in amyotrophic lateral sclerosis and frontotemporal dementia. Human molecular genetics. 2014;23(3): Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodriguez-Rodriguez E, Lopez de Munain A, et al. Assessing the role of the TREM2 p.r47h variant as a risk factor for Alzheimer's disease and frontotemporal dementia. Neurobiology of aging. 2014;35(2):444.e1-4. Cruchaga C, Karch CM, Jin SC, Benitez BA, Cai Y, Guerreiro R, et al. Rare coding variants in the phospholipase D3 gene confer risk for Alzheimer's disease. Nature. 2014;505(7484): Heckman MG, Elbaz A, Soto-Ortolaza AI, Serie DJ, Aasly JO, Annesi G, et al. Protective effect of LRRK2 p.r1398h on risk of Parkinson's disease is independent of MAPT and SNCA variants. Neurobiology of aging. 2014;35(1):266.e

92 Research Projects Code: Title: Follow-up genotyping and functional analisis of PSP H1 haplotype variants Principal Investigator: Pau Pastor CIBERNED s collaboration: CIBERNED s groups: Type: Others Funding Agency: CurePSP's Research Program Funding: Duration: Code: RYC Title: Medida de perfusión cerebral en patología usando la técnica de imagen por resonancia magnética Principal Investigator: María A. Fernández Seara CIBERNED s groups: Type: National Funding Agency: MICCIN - Ramón y Cajal Funding: Duration: Code: SAF R Title: Identificación de genes implicados en enfermedad de Parkinson y temblor esencial mediante secuenciación de nueva generación y análisis de haplotipos Principal Investigator: Pau Pastor CIBERNED s groups: Type: National Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: PI 13/02211 Title: Marcadores de neuroimagen de la enfermedad de Parkinson geneticamente determinada Principal Investigator: María A. Pastor CIBERNED s groups: Type: National Funding Agency: Fondo de Investigaciones de la Seguridad Social FIS Funding: Duration: 2014 PhD Dissertations Author: Marta Vidorreta Díaz de Cerio Title: Medida de perfusión cerebral en la enfermedad de Parkinson usando la técnica de imagen de perfusión por resonancia magnética arterial spin labeling Date: 27th June 2014 Supervisor: María Asunción Fernández Seara Program 1 86

93 Group José Rodríguez Álvarez Program 1 CONTACT DETAILS Group José Rodríguez Álvarez Group José Rodríguez Álvarez Group José Instituto Rodríguez de Neurociencias ÁlvarezGroup José Rodrígue Álvarez Group José Rodríguez Álvarez Group José Rodríguez Álvarez Universidad Group Autónoma José Rodríguez de Barcelona Álvarez Group Jos Rodríguez Álvarez Group José Rodríguez Álvarez Group José Rodríguez Álvarez Group José Edificio Rodríguez M Álvarez Grou Campus de Bellaterra José Rodríguez Álvarez Group José Rodríguez Álvarez Group Cerdanyola José Rodríguez del Valles, Álvarez Barcelona Group (Spain) José Rodríguez Álvare Group José Rodríguez Álvarez Group José Rodríguez Álvarez Group José Rodríguez Phone: +34 Álvarez Group 861 José Rodrígue jose.rodriguez@uab.es Álvarez Group José Rodríguez Álvarez Group José Rodríguez Álvarez Group José Rodríguez Álvarez Group José Rod LIST OF PERSONNEL José Rodríguez Álvarez Principal Investigator, Associate Professor José Aguilera Ávila Associate Professor Muriel Arimón Bedós PhD student Ana María Candalija Iserte PhD student Meng Chen PhD student Wenwen Cheng Postdoctoral Fellow Lilian Enriquez Barreto Postdoctoral Fellow Rut Fadó Andrés Postdoctoral Fellow Míriam Javier Torrent Postdoctoral Fellow Sergi Marco Martín Postdoctoral Fellow Alfredo Jesús Miñano Molina Postdoctoral Fellow Laura Ortega Hernandez Postdoctoral Fellow Guillem Sanchez Opazo PhD student Carlos Saura Antolín Associate Professor 87

94 Summary Multiple evidences have supported a role for beta amyloid (Aβ) in the etiology and pathogenesis of AD. It is believed that the progressive accumulation of self-aggregates of Aβ as oligomers (oaβ) would mediate synaptic dysfunction, leading to the initial cognitive deficits observed in MCI and earlier AD stages. Changes in gene expression programs and synaptic receptors and/or scaffolding proteins function could be underlying the early synaptic dysfunction associated to cognitive impairment in MCI and early AD. In the last 5 years we have reported that synaptic activity and memory training activate a transcriptional program dependent on the camp-responsive element binding protein (CREB)-regulated transcription coactivator-1 (CRTC1). During the last year we have reported that altered CRTC1 activation due to reduced dephosphorylation (Ser151) and nuclear translocation of CRTC1 results in transcriptional gene changes in transgenic mice expressing the β-amyloid precursor protein (APPSw,Ind) at early amyloid pathology and cognitive stages. In human brain CRTC1-dependent transcriptional changes are observed independently of CRTC1 protein levels at intermediate AD pathological stages. Finally, adenoviral-mediated CRTC1 gene transfer reverses Aβ-induced memory deficits by restoring the expression of a specific subset of CRTC1 target genes. On the other hand, we have been also investigating the role of oaβ on the levels of AKAP150, a synaptic scaffolding protein that also binds to GluA1 subunit-ampar. AKAP79/150 (human 79/rodent 150) is a scaffold protein encoded by the AKAP5 gene that targets PKA, PKC and CaN to the PSD to regulate AMPAR phosphorylation, trafficking, and activity associated with synaptic strength. Our data suggests that oaβ affects the AKAP79/150 anchoring ability that regulates CaN functions and could be related to a deregulation of synaptic proteins like GluA1 that are crucial for the synaptic function during learning and memory processes in the hippocampus. Keywords Alzheimer disease, Memory, Intracelular signaling, Gene regulation, Glutamate receptors, CRTC1, Early synaptic dysfunction Publications Blancas S, Fado R, Rodriguez-Alvarez J, Moran J. Endogenous XIAP, but not other members of the inhibitory apoptosis protein family modulates cerebellar granule neurons survival. International journal of developmental neuroscience: the official journal of the International Society for Developmental Neuroscience. 2014;37: Epub 2014 Jun 21. Xifro X, Minano-Molina AJ, Saura CA, Rodriguez-Alvarez J. Ras protein activation is a key event in activity-dependent survival of cerebellar granule neurons. The Journal of biological chemistry. 2014;289(12): Epub 2014 Feb 12. Candalija A, Cubi R, Ortega A, Aguilera J, Gil C. Trk receptors need neutral sphingomyelinase activity to promote cell viability. FEBS letters. 2014;588(1): Epub 2013 Dec 5. Parra-Damas A, Valero J, Chen M, Espana J, Martin E, Ferrer I, et al. Crtc1 activates a transcriptional program deregulated at early Alzheimer's disease-related stages. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(17): Balmaceda V, Cuchillo-Ibanez I, Pujadas L, Garcia-Ayllon MS, Saura CA, Nimpf J, et al. ApoER2 processing by presenilin-1 modulates reelin expression. FASEB journal: official publication of the Federation of American Societies for Experimental Biology. 2014;28(4): Program 1 88

95 2014 Annual Report José Rodríguez Álvarez Group Research Projects Code: 2014 SGR 0984 Title: Recerca Biomédica en Neurodegeneració (REBINE) Principal Investigator: José Rodríguez Álvarez CIBERNED s groups: Type: CCAA Funding Agency: AGAUR-Generalitat Catalunya Funding: Duration: Code: SAF Title: Regulación de la señalización mediada por los receptores con dominios de muerte por antagonistas y mirnas como estrategia protectora en la isquemia cerebral Principal Investigator: José Rodríguez Álvarez CIBERNED s groups: Type: International Funding Agency: Ministerio de Ciencia e Innovación Funding: Duration: Code: A S Title: Transcriptional mechanisms of memory loss in Alzheimer s disease Principal Investigator: Carlos A. Saura CIBERNED s groups: Type: International Funding Agency: BrightFocus Foundation Funding: Duration: Code: SAF Title: Transcriptional mechanisms underlying memory loss in Alzheimer s disease transgenic mouse models (TRANSMECAD) Principal Investigator: Carlos A. Saura CIBERNED s groups: Type: National Funding Agency: MINECO Funding: Duration: PhD Dissertations Author: Wen wen Cheng Title: Amyloid-? oligomers reduces glutamatergic transmission and inhibits synaptic activity with underlying mechanisms suggestting akap150 modulates diverse synaptic functions Date: 9th October 2014 Supervisor: José Rodríguez Álvarez Author: Guillem Sánchez Opazo Title: Modulació de la supervivencia i la diferenciació neuronals per les proteïnes inhibidores de l apoptosi Date: 18th September 2014 Supervisor: José Rodríguez Álvarez Author: Meng Chen Title: Role of the transcriptional coactivator CRTC1 on hippocampal-dependent associative memory Date: 1st October 2014 Supervisor: Carlos A. Saura Antolín 89

96 Group Javier Sáez Valero Program 1 CONTACT DETAILS oup Javier Sáez Valero Group Javier Sáez Valero Group Javier Instituto Sáez Valero de Neurociencias Group Javier de Sáez Alicante Valero Group Javier ez Valero Group Javier Sáez Valero Group Javier Sáez Valero Group Universidad Javier Sáez Miguel Valero Hernández-CSIC Group Javier Sáez Valero oup Javier Sáez Valero Group Javier Sáez Valero Group Javier Sáez Valero Avenida Group Ramón Javier y Sáez Cajal, Valero s/n Group Javier Sant Joan d Alacant (Spain) ez Valero Group Javier Sáez Valero Group Javier Sáez Valero Group Javier Phone: Sáez Valero Group Javier Sáez Valero oup Javier Sáez Valero Group Javier Sáez Valero Group Javier Sáez Valero Group Fax: +34 Javier 965 Sáez 919 Valero 561 Group Javier j.saez@umh.es ez Valero Group Javier Sáez Valero Group Javier Sáez Valero Group Javier Sáez Valero Group Javier Sáez Valero LIST OF PERSONNEL Javier Sáez Valero Principal Investigator, Associate Proffesor Inmaculada Cuchillo Ibáñez Postdoctoral Fellow Inmaculada López Font Postdoctoral Fellow Trinidad Mata Balaguer Postdoctoral Fellow María Salud García Ayllón Postdoctoral Fellow Jordi Alom Poveda Head of Neurology department Esther Llorens Álvarez Lab Technician Aitana Sogorb Esteve PhD student Valeria Balmaceda Valdez PhD student Maria Letizia Campanari PhD student 90

97 2014 Annual Report Javier Sáez Valero Group Summary The aim of our research group stills focus into Alzheimer s disease (AD) from a basic point of view, but also regarding translational benefits including clinical-diagnostic applications. In both, basic and translational, research lines we are focusing in the study of presenilin 1 (PS1). PS1 is the active component of the γ-secretase complex, which processes the amyloid precursor (APP) generating the β-amyloid or Aβ peptide, ultimately responsible for the development of AD. Many other substrates, besides APP, are potential substrates of PS1/γ-secretase, being relevant for the design of an appropriate therapeutic strategy. Recently, we have demonstrate that the Reelin receptor, ApoER2 (Balmaceda et al., 2014), and the acetylcholinesterase membrane-anchoring subunit PRiMA, are also substrates for PS1 (García-Ayllon et al., 2014). Reelin is a signaling protein increasingly associated with the pathogenesis of AD that relevantly modulates tau phosphorylation. We have previously demonstrated that the Aβ peptide alters Reelin expression. The interaction of Reelin with its membrane receptor ApoER2 (also receptor for ApoE) transduces the signal intracellular, and ultimately controls tau phosphorylation. The binding of Reelin to ApoER2 induces the cleavage of ApoER2 by γ-secretase generating an intracellular ApoER2 C-terminal fragment which, in turns, modulates the transcriptional activity of the Reelin promotor, revealing a complex regulation of Reelin signaling. Regarding the cholinergic enzyme acetylcholinesterase (AChE) we also demonstrated that cleavage of the membrane anchor PRiMA by γ-secretase results in a C-terminal PRiMA fragment. In this context, despite the overall decrease in AChE activity levels in AD, its protein content does not accord with the decrease in enzymatic activity level (Campanari et al., 2014). The potential significance of these unexpected levels of inactive AChE protein in the AD brain is intriguing, particularly in the context of proteinprotein interactions between AChE and PS1, being AChE able to modulate PS1 levels and γ-secretase activity (Campanari et al., 2014). Keywords Alzheimer s β-amyloid, Presenilin, Secretase, P-tau, Reelin, Acetylcholinesterase, Biomarker Publications Bi CW, Luk WK, Campanari ML, Liu YH, Xu L, Lau KM, et al. Quantification of the transcripts encoding different forms of AChE in various cell types: real-time PCR coupled with standards in revealing the copy number. Journal of molecular neuroscience: MN. 2014;53(3): Epub 2014 Jan 3. de-madaria E, del Mar Frances M, Gea-Sorli S, Gutierrez LM, Viniegra S, Perez-Mateo M, et al. Role of protease-activated receptor 2 in lung injury development during acute pancreatitis in rats. Pancreas. 2014;43: Epub Hallaq R, Volpicelli F, Cuchillo-Ibanez I, Hooper C, Mizuno K, Uwanogho D, et al. The Notch intracellular domain represses CRE-dependent transcription. Cellular signalling. [Epub ahead of print]. Epub 2014 Dec 3. Campanari ML, Garcia-Ayllon MS, Belbin O, Galceran J, Lleo A, Saez-Valero J. Acetylcholinesterase modulates presenilin-1 levels and γ-secretase activity. Journal of Alzheimer's disease: JAD. 2014;41(3): Garcia-Ayllon MS, Campanari ML, Montenegro MF, Cuchillo-Ibanez I, Belbin O, Lleo A, et al. Presenilin-1 influences processing of the acetylcholinesterase membrane anchor PRiMA. Neurobiology of aging. 2014;35(7): Epub 2014 Feb 6. 91

98 Balmaceda V, Cuchillo-Ibanez I, Pujadas L, Garcia-Ayllon MS, Saura CA, Nimpf J, et al. ApoER2 processing by presenilin-1 modulates reelin expression. FASEB journal: official publication of the Federation of American Societies for Experimental Biology. 2014;28(4): Campanari ML, Garcia-Ayllon MS, Blazquez-Llorca L, Luk WK, Tsim K, Saez-Valero J. Acetylcholinesterase protein level is preserved in the Alzheimer's brain. Journal of molecular neuroscience: MN. 2014;53(3): Research Projects Code: PI12/00593 Title: Alteración en la vía de señalización de Reelina en la enfermedad de Alzheimer: implicaciones patológicas Principal Investigator: Javier Sáez Valero CIBERNED s groups: Type: International Funding Agency: ISC-III Funding: Duration: Code: FPA/2014/048 Title: Ayuda para la contratación de personal de apoyo en centros de investigación de la Comunitat Valenciana Principal Investigator: Javier Sáez Valero CIBERNED s groups: Type: International Funding Agency: Dirección General de Universidad, Estudios Superiores y Ciencia, GVA Funding: Duration: 2014 Code: PI11/ Title: Biomarkers for Alzheimer s disease and Parkinson s disease (BIOMARKAPD) Principal Investigator: Javier Sáez Valero CIBERNED s collaboration: Yes CIBERNED s groups: G504; G407 Type: European Funding Agency: ISC-III Funding: Duration: Code: 23/14 Lote 1.1 Title: Equipo de Imagen en infrarrojo para detección simultánea de dos señales de fluorescencia Principal Investigator: Jorge Manzanares CIBERNED s groups: Type: Others Funding Agency: Universidad Miguel Hernández Funding: Duration: 2014 Code: Fundación Ramón Areces Title: Regulación epigenética de Reelina en la enfermedad de Alzheimer Principal Investigator: Javier Sáez Valero CIBERNED s groups: Type: International Funding Agency: Fundación Ramón Areces (XVI Concurso Nacional para la Adjudicación de Ayudas a la Investigación Científica y Técnica) Funding: Duration: Code: RGC Title: The trafficking of PRiMA-linked tetrameric AChE: Probing roles of glycosylation in determining the degradation of AChE in the brain Principal Investigator: Karl Tsim CIBERNED s groups: Type: International Funding Agency: Resarch Grant Council, Hong Kong Funding: Duration: Code: SEP Title: A natural compensation ovine model of a RELNknockout, opening new insights into compensation mechanisms to gain knowledge into mental disorders Principal Investigator: Beatriz Gutierrez-Gil (Universidad de León) CIBERNED s groups: Type: International Funding Agency: Horizon 2020, Excellent Science Funding: N.D. Duration: 2014 Program 1 92

99 2014 Annual Report Javier Sáez Valero Group PhD Dissertations Author: Valeria Balmaceda Valdez Title: La ruta de señalización de la reelina y su alteración en la enfermedad de Alzheimer Date: 14th April 2014 Supervisor: Javier Sáez Valero Author: Maria Letizia Campanari Title: Cross-talk between acetylcholinesterase and presenilin 1, implications for Alzheimer s disease Date: 29th April 2014 Supervisor: Javier Sáez Valero Author: Asunción Candela Gomis Title: Alteraciones de las colinesterasas en un modelo experimental de encefalopatía hepática en la rata cirrótica Date: 27th June 2014 Supervisor: Javier Sáez Valero 93

100 Group Eduardo Soriano García Program 1 CONTACT DETAILS oup Eduardo Soriano García Group Eduardo Soriano García Group Eduardo Soriano Universitat García de Group Barcelona Eduardo Soriano rcía Group Eduardo Soriano García Group Eduardo Soriano García Group Eduardo Parc Científic Soriano de Barcelona García Group Eduardo riano García Group Eduardo Soriano García Group Eduardo Soriano García Group Baldiri Eduardo Reixac, Soriano García Group Barcelona (Spain) Eduardo Soriano García Group Eduardo Soriano García Group Eduardo Soriano Phone: García Group Eduardo Soriano rcía Group Eduardo Soriano García Group Eduardo Soriano García Group Eduardo Soriano García Group Eduardo riano García Group Eduardo Soriano García Group Eduardo Soriano García Group esoriano@ub.edu Eduardo Soriano García LIST OF PERSONNEL Eduardo Soriano García Principal investigator, Full Professor Ferrán Burgaya Márquez Associate Professor, Researcher Jesús Ureña Bares Associate Professor, Researcher Marta Pascual Pérez Associate Professor, Researcher Fausto Ulloa Darquieta Postdoctoral Fellow Ashraf Mulhaisen Lab Technician Lluís Pujadas Coll Postdoctoral Fellow Tiziana Cotrufo Postdoctoral Fellow Daniela Rossi Postdoctoral Fellow Nuria Masachs Jahoner Postdoctoral Fellow Yasmina Manso Sanz Postdoctoral Fellow Serena Mirra Postdoctoral Fellow Antoni Parcerisas Ferrer PhD student Irene Lobon Pérez PhD student Marc Hernaiz Flores PhD student 94

101 2014 Annual Report Group Eduardo Soriano García Summary The main goal of the laboratory is to understand how key developmental genes play a fundamental role in neuronal plasticity in the adult brain, which is crucial for complex neural functions (eg., learning and memory). The rationale is that adult plasticity (adult neurogenesis and synaptic plasticity) is reminiscent of developmental processes. Because dysregulation of adult neurogenesis and synaptic plasticity are implicated in neurological, neurodegenerative and psychiatric disorders, we aim also to understand how these genes contribute to the pathology of these diseases and whether modelling developmental genes in the adult brain ameliorate these neural disorders. Reelin is an extracellular protein that is critical for neural migration and synaptogenesis. To unravel the function of this developmental gene specifically in the adult forebrain, we have generated conditional transgenic mice that overexpress Reelin. Using this tool, we have demonstrated that this gene is essential in adult plasticity (both adult neurogenesis and synaptic plasticity) and that its overexpression prevents amyloid burden and structural and cognitive deficits in Alzheimer Disease, and the development of psychiatric-like phenotypes, including cocaine addiction (Pujadas, 2010, 2014; Teixeira, 2011, 2012, 2013). Recently, we have recently generated conditional (floxed) reelin KO mice that are under analysis. Keywords Neural plasticity, Adult neurogenesis, Alzheimer Disease, Synaptogenesis Publications Barbosa DJ, Serrat R, Mirra S, Quevedo M, Gomez de Barreda E, Avila J, et al. MDMA impairs mitochondrial neuronal trafficking in a Tau- and Mitofusin2/Drp1-dependent manner. Archives of toxicology. 2014;88(8): Epub 2014 Feb 13. Gomez-Ramos A, Sanchez-Sanchez R, Muhaisen A, Rabano A, Soriano E, Avila J. Similarities and differences between exome sequences found in a variety of tissues from the same individual. PloS one. 2014;9(7):e Epub 2014 Jul 1. Parcerisas A, Rubio SE, Muhaisen A, Gomez-Ramos A, Pujadas L, Puiggros M, et al. Somatic signature of brain-specific single nucleotide variations in sporadic Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;42(4): Barbosa DJ, Serrat R, Mirra S, Quevedo M, de Barreda EG, Avila J, et al. The mixture of "ecstasy" and its metabolites impairs mitochondrial fusion/fission equilibrium and trafficking in hippocampal neurons, at in vivo relevant concentrations. Toxicological sciences: an official journal of the Society of Toxicology. 2014;139(2): Epub 2014 Mar 4. Avila J, Gomez-Ramos A, Soriano E. Variations in brain DNA. Frontiers in aging neuroscience. 2014;6:323. Epub 2014 Nov 25. Serra-Vidal B, Pujadas L, Rossi D, Soriano E, Madurga S, Carulla N. Hydrogen/deuterium exchangeprotected oligomers populated during Aβ fibril formation correlate with neuronal cell death. ACS Chem Biol. 2014;9(11): Epub September 29, Senkov O, Andjus P, Radenovic L, Soriano E, Dityatev A. Neural ECM molecules in synaptic plasticity, learning, and memory. Prog Brain Res. 2014;214: Epub Pujadas L, Rossi D, Andres R, Teixeira CM, Serra-Vidal B, Parcerisas A, et al. Reelin delays amyloidbeta fibril formation and rescues cognitive deficits in a model of Alzheimer's disease. Nat Commun. 2014;5:3443. Epub 06 March

102 Armendariz BG, Masdeu M, Soriano E, Urena JM, Burgaya F. The diverse roles and multiple forms of focal adhesion kinase in brain. Eur J Neurosci. 2014;40(11): Epub September 29, Serrat R, Mirra S, Figueiro-Silva J, Navas-Perez E, Quevedo M, Lopez-Domenech G, et al. The Armc10/ SVH gene: genome context, regulation of mitochondrial dynamics and protection against Aβ-induced mitochondrial fragmentation. Cell death & disease. 2014;5:e1163. Epub 2014 Apr 10. Balmaceda V, Cuchillo-Ibanez I, Pujadas L, Garcia-Ayllon MS, Saura CA, Nimpf J, et al. ApoER2 processing by presenilin-1 modulates reelin expression. FASEB journal: official publication of the Federation of American Societies for Experimental Biology. 2014;28(4): Vega-Flores G, Rubio SE, Jurado-Parras MT, Gomez-Climent MA, Hampe CS, Manto M, et al. The GABAergic septohippocampal pathway is directly involved in internal processes related to operant reward learning. Cerebral cortex (New York, N.Y.: 1991). 2014;24(8): Teixeira CM, Masachs N, Muhaisen A, Bosch C, Perez-Martinez J, Howell B, et al. Transient downregulation of Dab1 protein levels during development leads to behavioral and structural deficits: relevance for psychiatric disorders. Neuropsychopharmacology: official publication of the American College of Neuropsychopharmacology. 2014;39(3): Research Projects Code: PI11/0704 Title: Alteraciones en la conexión GABAérgica septohipocámpica en la enfermedad de Alzheimer. Implicaciones terapéuticas Principal Investigator: Marta Pascual CIBERNED s groups: Type: National Funding Agency: Ministerio de Sanidad y Consumo (IS- CIII) Funding: Duration: Code: PI2013_08-1 Title: La dinámica mitocondrial y mitofagia como dianas terapéuticas en las Enfermedades de Parkinson y Huntington Principal Investigator: Eduardo Soriano CIBERNED s collaboration: Yes CIBERNED s groups: G301; G207; G410; G109; G408 Type: National Funding Agency: ISCIII Funding: Duration: Code: PI13/02641 Title: La tirosina quinasa Ack1 en neurodegeneración y muerte neuronal Principal Investigator: Jesús Ureña Bares CIBERNED s groups: Type: International Funding Agency: ISCIII Funding: Duration: 2014 Code: 613/C/2013 Title: Lipotoxicity, hepatic steatosis and hepatocarcinoma: role of the ARMC10/ARMCX family of mitochondrial proteins Principal Investigator: Eduardo Soriano CIBERNED s groups: Other CIBER s collaboration: CIBER Fisiopatología de la Obesidad y Nutrición Type: National Funding Agency: Fundació la Marató de TV3 Funding: Duration: Code: PI12/02108 Title: Participación de las Semaforinas transmembranales y la cinasa Fak en patologías relacionadas con la sinapsis y la actividad neuronal Principal Investigator: Ferrán Burgaya Márquez CIBERNED s groups: Type: International Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: COST Action BM1001: ECMNET Title: Brain Extracellular Matrix in Health and Disease Program 1 96

103 2014 Annual Report Eduardo Soriano García Group Principal Investigator: Alexander Dityatev CIBERNED s collaboration: Yes CIBERNED s groups: G408; G111 Type: International Funding Agency: COST (European Coordination in Science and Technology) European Office Funding: N.D. Duration: Code: SAF R Title: Nuevas Aproximaciones a la Patogénesis de la Enfermedad de Alzheimer Principal Investigator: Eduardo Soriano García CIBERNED s groups: Type: International Funding Agency: Ministerio de Economía y Competitividad Funding: N.D. Duration: PhD Dissertations Author: Carles Bosch Piñol Title: Role of Reelin in synaptogenesis and synaptic stabilization in the adult brain Date: 25th September 2014 Supervisor: Eduardo Soriano García Author: Maria del Mar Masdeu Camara Title: Papel de la tirosina quinasa Ack1 en la neuritogénesis y señalización regulada por neurotrofinas y moléculas de guía axonal Date: 30th May 2014 Supervisor: Eduardo Soriano García Author: Núria Masachs Janoher Title: Implicació de la via de senyalització de la Reelina en els transtorns psiquiàtrics i la neurogènesi adulta Date: 22nd October 2014 Supervisor: Eduardo Soriano García Author: Pablo José Barrecheguren Manero Title: Papel de las proteínas SNARE en guía axonal y progresión tumoral Date: 18th December 2014 Supervisor: Eduardo Soriano García 97

104 Group Ignacio Torres Alemán Program 1 CONTACT DETAILS oup Ignacio Torres Alemán Group Ignacio Torres Alemán Group Ignacio Torres Alemán Group Instituto Ignacio Cajal Torres Alemán oup Ignacio Torres Alemán Group Ignacio Torres Alemán Group Ignacio Avenidada Torres Alemán Doctor Arce, Group 37 Ignacio Torres emán Group Ignacio Torres Alemán Group Ignacio Torres Alemán Group Ignacio Torres Madrid Alemán (Spain) Group Ignacio Phone: rres Alemán Group Ignacio Torres Alemán Group Ignacio Torres Alemán Group Fax: +34 Ignacio 915 Torres Alemán Group nacio Torres Alemán Group Ignacio Torres Alemán Group Ignacio Torres Alemán Group torres@cajal.csic.es Ignacio Torres Alemán Grup emán Group Ignacio Torres Alemán Group Ignacio Torres Alemán Group Ignacio Torres Alemán Group Ignacio Tor LIST OF PERSONNEL Ignacio Torres Alemán Principal Investigator Manuel Domínguez Researcher Ana M Fernández Researcher Miguel García Researcher Laura Genis Researcher Dolores Guinea IResearcher Edwin Hernández Researcher Victor Munive Researcher Andrea Santi Researcher Angel Trueba Researcher 98

105 2014 Annual Report Ignacio Torres Alemán Group Summary The lab is focused on the role of insulin factors in the healthy and diseased brain. In 2014 we continued studying a little explored concept: the role of insulin and IGF-I in regulating brain glucose metabolism and underlying mechanisms that include glucose transporter 1. This new role of these hormones will in all probability modify our understanding of the biological significance of insulin peptides in the adult brain. Also within a physiological context, we are analyzing the role of IGF-I in neuroprotection by physical exercise and the remarkable gender differences observed due to the modulatory actions of estrogen. In addition, we are interested in unveiling mechanisms of serum-to-brain traffic of circulating IGF-I under physiological and pathological conditions. We are currently investigating the effect of diet on this traffic and plan to incorporate the analysis of stress in the near future. Regarding pathology we are interested in neuro-inflammation and its links with Alzheimer s disease within a CIBERNED collaborative program. We are also analyzing the role of IGF-I in the responses to oxidative stress by astrocytes. In this regard, we are now determining a new ligand-independent role of the IGF-I receptor. Recently we have incorporated posttraumatic stress syndrome as a pathology of great interest because of its current clinical impact and because it possibly involves many of the known neuroprotective actions of IGF-, from protection against physicial injury and oxidative stress to modulation of mood and cognition. Keywords Role of Insulin like growth factors in neuronal plasticity and degenerative diseases, Therapeutic targets in neurodegeneration, The aging brain and cognition, Blood brain barriers Publications Genis L, Davila D, Fernandez S, Pozo-Rodrigalvarez A, Martinez-Murillo R, Torres-Aleman I. Astrocytes require insulin-like growth factor I to protect neurons against oxidative injury. F1000Research. 2014;3:28. Epub 2014 Jan 28. Sainz-Gallego I, Rodriguez de Rivera FJ, Pulido I, Torres-Aleman I, Arpa J. IGF-1 in autosomal dominant cerebellar ataxia- open-label trial. Cerebellum and Ataxias. 2014;1(13):1-9. Sainz-Gallego I, Torres-Aleman I, Arpa J. IGF-1 in Friedreich s ataxia: proof of concept Trial. Cerebellum and Ataxias. 2014;1(10):1-8. Fernandez S, Genis L, Torres-Aleman I. A phosphatase-independent gain-of-function mutation in PTEN triggers aberrant cell growth in astrocytes through an autocrine IGF-1 loop. Oncogene. 2014;33(32): Research Projects Code: S2010_BMD Title: Redes de señalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas Principal Investigator: José González Castaño CIBERNED s collaboration: Yes CIBERNED s groups: G401; G104; G111; G307; G409; G412 Other CIBER s collaboration: Yes (CIBERER) Type: International Funding Agency: Comunidad de Madrid Funding: Duration:

106 Code: PI2013/01 Title: Propiedades emergentes de la relación neurona-glía que subyacen a neurodegeneración y demencia en la enfermedad de Alzheimer Principal Investigator: Ignacio Torres Alemán CIBERNED s collaboration: Yes CIBERNED s groups: G413; G204; G409; G108; G411; G415 Type: National Funding Agency: CIBERNED Funding: Duration: Code: SAF R Title: Interacción del receptor IGF-I con co-receptores y su impacto en la demencia de Alzheimer Principal Investigator: Ignacio Torres Alemán CIBERNED s groups: Type: National Funding Agency: MINECO Funding: Duration: 2014 Code: Inter-CIBERs Title: Molecular links between diabetes and neurodegenerative disorders Principal Investigator: Angel Raya CIBERNED s collaboration: Yes CIBERNED s groups: G409; G604 Other CIBER s collaboration: Yes (CIBERDEM) Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: 2014 PhD Dissertations Author: Edwin Hernández Garzon Title: Regulación de la captura de glucosa por astrocitos por factores insulínicos Date: 12nd December 2014 Supervisor: Ignacio Torres Alemán Program 1 100

107 Group Ramón Trullás Oliva Program 1 CONTACT DETAILS Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Unidad Oliva Group de Neurobiología Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás IIBB-CSIC Oliva Group Ramó IDIBAPS Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Rosselló Ramón 161, 7ª Trullás Planta, Oliva Lab. Group 711 Ramón Trullá Barcelona (Spain) Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Phone: Trullás +34 Oliva 659 Group Ramón Trullás Oliv Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Fax: Group Ramón Trullás Oliva Grou ramon.trullás@iibb.csic.es Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group Ramón Trullás Oliva Group LIST OF PERSONNEL Ramón Trullás Oliva Principal Investigator, CSIC Research Professor Nicole Mahy Professor of Biochemistry Manuel Rodríguez Assistant Professor Petar Podlesniy Research Scientist Nuria Serra Technician Laura Mañas Technician Margalida Puigròs PhD student 101

108 Summary During the past year we have investigated the influence of Neuronal Pentraxin 1 (NP1) on mitochondrial dynamics and synaptogenesis. We have shown that NP1, a pro-apoptotic protein that is increased in dystrophic neurites in brains with Alzheimer s disease (AD), decreases the number of excitatory synapses and inhibits neuronal plasticity and long term potentiation. At the same time we have observed that NP1 alters the balance between mitochondrial fusion and fission processes. We are also studying the mechanisms by which NP1 alters mitochondrial dynamics and replication of mitochondrial DNA (mtdna) in models of AD. In addition, we have continued the studies investigating the relationship between the content of circulating extracellular mtdna in cerebrospinal fluid (CSF) and various neurodegenerative diseases. We have confirmed that a decrease of mtdna in the CSF occurs in clinical and pre-clinical forms of Alzheimer s disease, but not in other neurodegenerative disorders such as Creutzfeld-Jakob disease or other dementias associated with various neurodegenerative disorders different from AD. We have also confirmed that a decrease of mtdna content in CSF precedes the onset of clinical signs of sporadic AD. These results provide new evidence for the hypothesis that mtdna depletion is a fundamental pathophysiological mechanism of AD and that restoration of mtdna copy number to normal levels should have a therapeutic effect and prevent AD neurodegeneration. Keywords Synaptic Neurodegeneration, Alzheimer s Disease Biomarkers, Apoptosis, Mitochondrial dynamics, Mitochondrial transport in neurodegeneration, Neuronal Death, Neuronal Pentraxins, Molecular mechanisms of Alzheimer s disease Publications Popik P, Holuj M, Nikiforuk A, Kos T, Trullás R, Skolnick P. 1-Aminocyclopropanecarboxylic acid (ACPC) produces procognitive but not antipsychotic-like effects in rats. Psychopharmacologia. [Epub ahead of print]. Epub 2014 Sep 27. Podlesniy P, Figueiro-Silva J, Llado A, Antonell A, Sanchez-Valle R, Alcolea D, et al. Characterization of mtdna detection in cerebrospinal fluid. Reply: To PMID Annals of neurology. 2014;75(3): Epub 2014 Mar 7. Lopez-Lopez A, Gamez J, Syriani E, Morales M, Salvado M, Rodriguez MJ, et al. CX3CR1 is a modifying gene of survival and progression in amyotrophic lateral sclerosis. PloS one. 2014;9(5):e Epub 2014 May 7. Gimeno-Bayon J, Lopez-Lopez A, Rodriguez MJ, Mahy N. Glucose pathways adaptation supports acquisition of activated microglia phenotype. Journal of neuroscience research. 2014;92(6): Epub 2014 Feb 7. Serrat R, Mirra S, Figueiro-Silva J, Navas-Perez E, Quevedo M, Lopez-Domenech G, et al. The Armc10/ SVH gene: genome context, regulation of mitochondrial dynamics and protection against Aβ-induced mitochondrial fragmentation. Cell death & disease. 2014;5:e1163. Epub 2014 Apr 10. Espinosa-Parrilla JF, Martinez-Moreno M, Gasull X, Mahy N, Rodriguez MJ. The L-type voltagegated calcium channel modulates microglial pro-inflammatory activity. Molecular and cellular neurosciences. 2015;64: Epub 2014 Dec 12. Lamarca A, Gella A, Martianez T, Segura M, Figueiro-Silva J, Grijota-Martinez C, et al. Uridine 5'-triphosphate promotes in vitro Schwannoma cell migration through matrix metalloproteinase-2 activation. PloS one. 2014;9(6):e Epub 2014 Jun 6. Program 1 102

109 2014 Annual Report Ramón Trullás Oliva Group Research Projects Code: SAF Title: Mechanisms of synapse elimination and mitochondrial dynamics in neurodegeneration: role of Neuronal Pentraxin 1 Principal Investigator: Ramón Trullás Oliva CIBERNED s groups: Type: National Funding Agency: MICINN Funding: Duration: Code: PI2013_08-1 Title: La dinámica mitocondrial y mitofagia como dianas terapéuticas en las Enfermedades de Parkinson y Huntington Principal Investigator: Eduardo Soriano CIBERNED s collaboration: Yes CIBERNED s groups: G301; G207; G410; G109; G408 Type: National Funding Agency: ISCIII Funding: Duration:

110 Group Francisco Javier Vitorica Ferrández Program 1 CONTACT DETAILS oup Francisco Javier Vitorica Ferrández Group Francisco Javier Dpto. Vitorica Bioquímica Ferrández y Biología Group Molecular Francisco Javier Vitorica rrández Group Francisco Javier Vitorica Ferrández Group Francisco Javier Vitorica Facultad Ferrández Group Farmacia Francisco Javier Universidad de Sevilla torica Ferrández Group Francisco Javier Vitorica Ferrández Group Francisco Javier Vitorica Ferrández Group Francisco Instituto de Biomedicina de Sevilla vier Vitorica Ferrández Group Francisco Javier Vitorica Ferrández Hospital Group Universitario Francisco Javier Virgen Vitorica del Rocío Ferrández Group ancisco Javier Vitorica Ferrández Group Francisco Javier Vitorica Ferrández Group Francisco Sevilla Javier (Spain) Vitorica Ferrández Phone: oup Francisco Javier Vitorica Ferrández Group Francisco Javier Vitorica Ferrández Fax: Group Francisco Javier Vitorica vitorica@us.es rrández Group Francisco Javier Vitorica Ferrández Group Francisco Javier Vitorica Ferrández Group Francisco Javier LIST OF PERSONNEL Francisco Javier Vitorica Ferrández Principal Investigator, Full Professor Marisa Vizuete Chacón University Professor Victoria Navarro Garrido Fellow FPI Francisco Javier Moyano Fellow FPU Mª Virtudes Sánchez Mico Fellow FPU Sebastian Jiménez Muñoz Technician Mª Luisa García-Calvo Technician 104

111 2014 Annual Report Francisco Javier Vitorica Ferrández Group Summary During 2014 our group has continued with the characterization of the inflammatory response in the hippocampus, using human postmortem samples with different Braak stages; from BraakII to Braak V-VI, the latter are patients diagnosed as Alzheimer type of dementia. To perform this characterization, we have first determined, the load of both Abeta and phosphorylated tau, as well as the neurodegeneracion in this cohort. Second, the inflammatory response was characterized by determining the expression of mrna/protein levels or by immunohistochemistry (in collaboration with the group of Dr. A. Gutierrez, University of Málaga) of genes considered key for microglial activation and/or differentiation. This activity is part of the objectives of a collaborative project from CIBERNED, developed during Also within this context, we isolated the so-called soluble fractions (extracellular and cytosolic) of different human samples. The content of soluble Abeta and/or tau forms was analyzed by western blots, immunoprecipitations and ELISAs. Furthermore, we have analyzed, in vitro, the toxicity and/or immunogenicity of these fractions. Finally, these soluble fractions were characterized using proteomic techniques (in collaboration with Dr. J Vazquez, CNIC). On the other hand, we are also determining the role of the microglial activation in the evolution of the pathology of Alzheimer, using APP or PS1xAPP models. Specifically, using transgenic models, the microglial response is being pharmacological/genetically manipulated (inhibited or enhanced) and the effect on the pathology (accumulation of Abeta, tau phosphorylation, neurodegeneration) was analyzed. In late 2014 we have initiated a new collaborative project (coordinated by Dr. E Galea) investigating the properties of active astrocytes and their role in the development of Alzheimer pathology. This project is funded by the Marató from TV3. Moreover, for the characterization of the PS1xAPP transgenic models, we have established a new collaboration with Dr. JL Gómez-Ariza (University of Huelva) to study, from a metabolomic approach, the evolution of the pathology in the transgenic mice. Finally, we have also requested, together with the company Neuron Biopharma, a collaborative project from Retos-Colaboración call, promoted by the Ministry of Economy and Finance. Our collaboration in this project is based on the preclinical characterization of first-in-class molecules as modifiers of neurological/degenerative diseases. This project has a marked clinical translational character. Keywords Alzheimer, Neurodegeneration, Neuroprotection, Inflammation, Abeta, Oligomers, Transgenic models, Neuropathology Publications Gonzalez-Dominguez R, Garcia-Barrera T, Vitorica J, Gomez-Ariza JL. Metabolomic screening of regional brain alterations in the APP/PS1 transgenic model of Alzheimer's disease by direct infusion mass spectrometry. Journal of pharmaceutical and biomedical analysis. 2014;102C: Epub 2014 Oct 22. Gonzalez-Dominguez R, Garcia-Barrera T, Vitorica J, Gomez-Ariza JL. Metabolomics reveals significant impairments in the immune system of the app/ps1 transgenic mice of alzheimer's disease. Electrophoresis. [Epub ahead of print]. Epub 2014 Nov

112 Gonzalez-Dominguez R, Garcia-Barrera T, Vitorica J, Gomez-Ariza JL. Region-specific metabolic alterations in the brain of the APP/PS1 transgenic mice of Alzheimer's disease. Biochimica et biophysica acta. 2014;1842(12 Pt A): Epub 2014 Oct 2. Jimenez S, Navarro V, Moyano J, Sanchez-Mico M, Torres M, Davila JC, et al. Disruption of amyloid plaques integrity affects the soluble oligomers content from Alzheimer disease brains. PloS one. 2014;9(12):e Epub 2014 Dec 8. Trujillo-Estrada L, Davila JC, Sanchez-Mejias E, Sanchez-Varo R, Gomez-Arboledas A, Vizuete M, et al. Early neuronal loss and axonal/presynaptic damage is associated with accelerated amyloid-β accumulation in AβPP/PS1 Alzheimer's disease mice subiculum. Journal of Alzheimer's disease: JAD. 2014;42(2): Cardona C, Sanchez-Mejias E, Davila JC, Martin-Rufian M, Campos-Sandoval JA, Vitorica J, et al. Expression of Gls and Gls2 glutaminase isoforms in astrocytes. Glia. 2015;63(3): Epub 2014 Oct 9. Jimenez AJ, Rodriguez-Perez LM, Dominguez-Pinos MD, Gomez-Roldan MC, Garcia-Bonilla M, Ho- Plagaro A, et al. Increased levels of tumour necrosis factor alpha (TNFα) but not transforming growth factor-beta 1 (TGFβ1) are associated with the severity of congenital hydrocephalus in the hyh mouse. Neuropathology and applied neurobiology. 2014;40(7): Torres M, Price SL, Fiol-Deroque MA, Marcilla-Etxenike A, Ahyayauch H, Barcelo-Coblijn G, et al. Membrane lipid modifications and therapeutic effects mediated by hydroxydocosahexaenoic acid on Alzheimer's disease. Biochimica et biophysica acta. 2014;1838(6): Research Projects Code: CTS 2035 Title: Oligomerización y toxicidad de los péptidos de Abeta: búsqueda de nuevas dianas de interes terapéutico en la enfermedad de Alzheimer Principal Investigator: Javier Vitorica CIBERNED s collaboration: Yes CIBERNED s groups: G415; G411 Type: CCAA Funding Agency: Junta de Andalucía Funding: Duration: Code: PI12/01439 Title: Oligómeros tóxicos del Abeta como agentes causantes de la disfunción del citoesqueleto y los procesos proteolíticos en la Principal Investigator: Javier Vitorica CIBERNED s collaboration: Yes CIBERNED s groups: G415; G411 Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: PI12/01431 Title: Oligómeros tóxicos del Abeta como agentes causantes de la disfunción del citoesqueleto y los procesos proteolíticos en la Principal Investigator: Antonia Gutiérrez Pérez CIBERNED s collaboration: Yes CIBERNED s groups: G411; G415 Type: International Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: PI2013/01 Title: Propiedades emergentes de la relación neurona-glía que subyacen a neurodegeneración y demencia en la enfermedad de Alzheimer Principal Investigator: Ignacio Torres-Alemán CIBERNED s collaboration: Yes CIBERNED s groups: G413; G204; G409; G108; G411; G415 Type: National Funding Agency: CIBERNED Funding: Duration: Program 1 106

113 Group Francisco Wandosell Jurado Program 1 CONTACT DETAILS Group Francisco Wandosell Jurado Group Francisco Centro Wandosell de Biología Jurado Molecular Group Severo Francisco Ochoa, Wandosell CSIC-UAM Jurado Group Fran cisco Wandosell Jurado Group Francisco Wandosell Jurado Group Francisco Universidad Wandosell Autónoma Jurado de Madrid Group Francisco Wan dosell Jurado Group Francisco Wandosell Jurado Group Francisco Wandosell Nicolás Jurado Cabrera Group 1, Cantoblanco Francisco Wandosell Jura Madrid (Spain) do Group Francisco Wandosell Jurado Group Francisco Wandosell Jurado Phone: Group Francisco /4591 Wandosell Jurado Grou Francisco Wandosell Jurado Group Francisco Wandosell Jurado Group Francisco Fax: Wandosell Jurado 420Group Francisc fwandosell@cbm.csic.es Wandosell Jurado Group Francisco Wandosell Jurado Group Francisco Wandosell Jurado Group Francisco Wandosel LIST OF PERSONNEL Francisco Wandosell Jurado Investigator, CSIC Staff Scientist Inés M. Antón CSIC Staff Scientist-Researcher Mª José Benítez Moreno Associate Professor María José Pérez Alvarez Assistant Professor UAM Juan José Garrido Jurado CSIC Staff Scientist-Researcher Lara Ordoñez Gutierrez Postdoctoral Fellow Jorge Cabrera Postdoctoral Fellow Ricardo Gargini Postdoctoral Fellow Laura Mateos-Montejo Postdoctoral Fellow Maria Isabel Escoll PhD student Irene Benito PhD student 107

114 Summary Our group, Molecular mechanisms of Neurodegeneration, is generated from an established line in the CBM over several years. At present, this CIBERNED line is formally composed by three sub-groups (PI s- F. Wandosell (CBM), I. Anton (CNB) and J.J. Garrido, Cajal Institute). Our group mostly interested in neurodegenerative disorders is now focusing in a group of brain disorders associated with aging, such as Alzheimer disease, Stroke and some type of brain tumours. In fact, we could consider that life-span or longevity is in some way specie-specific, and in addition can be affected by environment changes or mutations in simple genes. Thus one open question is... are there common molecular mechanisms underlying the age-dependency of several disorders? Data from different animal models strongly support that idea that longevity is controlled by several genetic pathways, such as: IGF1/Insulin-PI3K-Akt-FoxO. Some reports indicate that insulin and IGF-1 regulate longevity in a conserved manner. IGF-1/Insulin through their tyrosine kinase receptors controls the activity of Class1 phosphatidylinositol 3-kinase (PI3K) and several serine-threonine kinases such as Akt. We are interested in the analysis of the molecular mechanisms of some brain disorders, mainly focusing in the role of PI3K-Akt and some of the Akt substrates, trying to understand the key points of these processes. Our initial studies were focused on GSK3 and in Akt, we defined the role of GSK3 and Akt isoforms in the neuronal morphogenesis and how they are modified after Ischemia. We defined how the neuroprotective role of neurosteroids, such as estradiol, is due to the activation of PI3K-Akt pathway, at least in part (Co-PIs: J.J.Garrido & F. Wandosell). Second, we are analysing the role of the Akt downstream element, mtorc1. The kinase activity of this protein complex control, at least in part, protein synthesis and autophagy; whereas its dysfunction has been considering a key factor of beta amyloid generation and/or degradation in Alzheimer disease. Third, we are analysing some Akt-downstream elements (such as FoxO and Bim), and our data indicated that they are responsible of the cell division of cancerstem cells and the maintenance of its phenotype. Our work is now focusing on the conversion from astrocyte-astrocytoma-glioma triyng to define the role Akt-downtream elements and the regulation of cytoskeleton/cellular function (Co-PIs: I. Anton & F. Wandosell). In summary, we are focusing on track PI3K-Akt signalling and elements that control some physiological process, from cell division to differentiation, and how some of these proteins are modified in pathology. Keywords Neurodegeneration, Cellular Signaling, Neuroprotection, Estrogen, Alzheimer Disease, Neuritogenesis- Axonal Polarity, Cytoskeletal proteins, Actin Program 1 108

115 2014 Annual Report Francisco Wandosell Jurado Group Publications Vergara C, Ordonez-Gutierrez L, Wandosell F, Ferrer I, Del Rio JA, Gavin R. Role of PrPC Expression in Tau Protein Levels and Phosphorylation in Alzheimer's Disease Evolution. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Jun 26. Gargini R, Cerliani JP, Escoll M, Anton IM, Wandosell F. Cancer Stem Cell-Like Phenotype and Survival are Coordinately Regulated Byakt/Foxo/Bim Pathway. Stem cells (Dayton, Ohio). [Epub ahead of print]. Epub 2014 Nov 19. Perez-Alvarez MJ, Mateos L, Alonso A, Wandosell F. Estradiol and Progesterone Administration After pmcao Stimulates the Neurological Recovery and Reduces the Detrimental Effect of Ischemia Mainly in Hippocampus. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Nov 7. Wandosell F. Mecanismos moleculares de la enfermedad de Alzheimer: Causas genéticas y esporádicas. En: Garcia Rodriguez JC, Ed. Neuroprotección en enfermedades Neuro y Heredo degenerativas p Ordonez-Gutierrez L, Anton M, Wandosell F. Peripheral Amyloid Levels Present Gender Differences Associated with Aging in AβPP/PS1 Mice. Journal of Alzheimer's disease: JAD. [Epub ahead of print]. Epub 2014 Nov 14. Ordonez-Gutierrez L, Re F, Bereczki E, Ioja E, Gregori M, Andersen AJ, et al. Repeated intraperitoneal injection of liposomes containing phosphatidic acid and cardiolipin reduce amyloid-ß levels in APP/ PS1 transgenic mice. Nanomedicine: nanotechnology, biology, and medicine. [Epub ahead of print]. Epub 2014 Nov 14. Simon D, Herva ME, Benitez MJ, Garrido JJ, Rojo AI, Cuadrado A, et al. Dysfunction of the PI3K- Akt-GSK-3 pathway is a common feature in cell culture and in vivo models of prion disease. Neuropathology and applied neurobiology. 2014;40(3): Research Projects Code: SAF C03-01 Title: Papel de la vía PI3K/ AKT/mTOR en neurodegeneración Principal Investigator: Francisco Wandosell CIBERNED s groups: Type: International Funding Agency: MINECO Funding: Duration: 2014 Code: S2010_BMD Title: Redes de señalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas Principal Investigator: José González Castaño CIBERNED s collaboration: Yes CIBERNED s groups: G401; G104; G111; G307; G409; G412 Other CIBER s collaboration: Yes (CIBERER) Type: International Funding Agency: Comunidad de Madrid Funding: Duration:

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117 Parkinson s Disease, Huntignton s Chorea and other Movement Disorders Program 2 Group Jordi Alberch Vié Group Enric Isidre Canela Campos Group Valentín Ceña Callejo Group Antonio Cuadrado Pasto upo José Antonio del Río Fernández Group Isabel Fariñas Gómez Group Javier Fernández Ruiz Group José Manuel Fuente Rodríguez Group José Manuel García Verdugo Group José González Castaño Group Manuel Guzmán Pastor Group Teres Iglesias Vacas Group Jaime Kulisevsky Bojarski Group José Luis Labandeira García Group José Luis Lanciego Pérez Group Jos López Barneo Group José Javier Lucas Lozano Group María Ángeles Mena Gómez Group Rosario Moratalla Villalba Group José Ramón Naranjo Orovio Group José Ángel Obeso Inchausti Group Ana María Pérez Castillo Group Jordi Pérez Tur Grou Manuel Rodríguez Díaz Group Eduardo Tolosa Sarró Group Ignacio Torres Alemán Group Carlos Vicario Abejón Group Miquel Vila Bover Group Jordi Alberch Vié Group Enric Isidre Canela Campos Group Valentín Ceña Callejo Group Antoni Cuadrado Pastor Group José Antonio del Río Fernández Group Isabel Fariñas Gómez Group Javier Fernández Ruiz Group José Manuel Fuentes Rodríguez Group José Manuel García Verdugo Group José González Castaño Group Manuel Guzmá Pastor Group Teresa Iglesias Vacas Group Jaime Kulisevsky Bojarski Group José Luis Labandeira García Group José Lu Lanciego Pérez Group José López Barneo Group José Javier Lucas Lozano Group María Ángeles Mena Gómez Group Rosari Moratalla Villalba Group José Ramón Naranjo Orovio Group José Ángel Obeso Inchausti Group Ana María Pérez Castill Group Jordi Pérez Tur Group Manuel Rodríguez Díaz Group Eduardo Tolosa Sarró Group Ignacio Torres Alemán Group Carlo Vicario Abejón Group Miquel Vila Bover Group Jordi Alberch Vié Group Enric Isidre Canela Campos Group Valentín Ceñ Callejo Group Antonio Cuadrado Pastor Group José Antonio del Río Fernández Group Isabel Fariñas Gómez Group Javie Fernández Ruiz Group José Manuel Fuentes Rodríguez Group José Manuel García Verdugo Group José González Castañ Group Manuel Guzmán Pastor Group Teresa Iglesias Vacas Group Jaime Kulisevsky Bojarski Group José Luis Labandeir García Group José Luis Lanciego Pérez Group José López Barneo Group José Javier Lucas Lozano Group María Ángeles Men Gómez Group Rosario Moratalla Villalba Group José Ramón Naranjo Orovio Group José Ángel Obeso Inchausti Group An María Pérez Castillo Group Jordi Pérez Tur Group Manuel Rodríguez Díaz Group Eduardo Tolosa Sarró Group Ignacio Torre Research Groups Alberch Vié, Jordi Canela Campos, Enric Isidre Ceña Callejo, Valentín Cuadrado Pastor, Antonio del Río Fernández, José Antonio Fariñas Gómez, Isabel Fernández Ruiz, Javier Fuentes Rodríguez, José Manuel García Verdugo, José Manuel González Castaño, José Guzmán Pastor, Manuel Iglesias Vacas, Teresa Kulisevsky Bojarski, Jaime Labandeira García, José Luis Lanciego Pérez, José Luis López Barneo, José Lucas Lozano, José Javier Mena Gómez, María Ángeles Moratalla Villalba, Rosario Naranjo Orovio, José Ramón Obeso Inchausti, José Ángel Pérez Castillo, Ana María Pérez Tur, Jordi Rodríguez Díaz, Manuel Tolosa Sarró, Eduardo Vicario Abejón, Carlos Vila Bober, Miquel

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119 2014 Annual Report Scientific Report Program 2 CIBERNED s Program 2 brings together 27 basic and clinical research groups with a mainly translational character, joining forces to study neurodegenerative diseases of different etiology that cause important problems in patient s mobility. Among this group of diseases we can find, by decreasing prevalence: Parkinson s disease, Huntington s chorea, and different kinds of ataxias among other movement disorders. Research Line 1: Parkinson s Disease In Spain, 70,000 persons are affected by Parkinson s disease (PD), which is characterized by neuronal loss, the formation of Lewy bodies and neurites in the substantia nigra and the consequent loss of striatal dopamine (DA). However, it is currently well known that PD is a multisystem neurodegenerative process, in which, as the neurodegenerative process evolves, many areas of the nervous system are affected and there exists a deficit in several neurotransmission and neuromodulation systems. Preventing and correcting DA deficit are still important goals, but nowadays they are not considered to be the ultimate challenge in PD. Within this area, it is considered of vital importance to make progress on defining the following: a) Key aspects related to the ethiopathogenesis in PD. b) Physiopatological mechanisms related to disease onset and progression. c) Development of symptomatic treatments, especially neuroprotective and curative. Thus, a truly translational research is sought, having the disease and the patients as the main targets. The main research topics in this line are the following: Cognitive impairment and non-motor problems in PD Biomarkers in Parkinson s disease Problems related to symptomatic treatment: diskynesias New targets and novel therapeutic strategies in Parkinson s disease Circuits and physiopathology of basal ganglia Neuronal stress, cell protection and death in Parkinson s disease Neurogenesis and cell therapy in Parkinson s disease Early biomarkers in Parkinson s disease. Research Line 2: Huntington s disease and ataxias This program also focuses on research into other movement disorders such as Huntington s disease (HD) and ataxias. The prevalence of HD is much lower (about 4,500 patients in Spain) and it is characterized by the initial loss of spiny interneurons of the striatum. It is an autosomal dominant neurodegenerative pathology with complete penetrance produced by polyglutamine expansion in the huntingtin N-terminus. HD has no treatment and leads to death in around years depending on the number of polyglutamines, the age of onset, some unknown environmental factors and the modulation of some genes, some of which have been located but remain unidentified. The estimated prevalence of HD is 10 cases for every 100,000 persons, which means 4,500 patients in Spain and about 50,000 in the European Union. Its social health cost is considerable because of the importance of cognitive and motor deficits, as well as the severe behavioral problems that patients present. There is a large number of studies with neuroprotective drugs which modify the supposed pathogenic mechanisms or that are used in other neurodegenerative diseases. These drugs include inhibitors of neuronal excitation, coenzymes of the respiratory chain, vitamins, antioxidants, co-adjuvants in energy 113

120 production, etc. Some of these products offer encouraging results in experimental models of the disease but, unfortunately, they are not later confirmed in the clinic. The study of HD is important because is the best studied and most prevalent model of neurodegenerative diseases caused by triplet expansions, which also includes some ataxias. Discovering the pathogenic mechanisms of HD and finding an effective, either neuroprotective or curative, would have immediate implications on any of the other neurodegenerative pathologies caused by triplet expansions. The main research topics in this line are the following: Identification of molecular and cellular basis of Huntington s disease Experimental studies in animal models of Huntington s disease Clinic, genetics and neuropathology of Huntington s disease Program 2 114

121 2014 Annual Report Scientific Report Groups Principal Investigator Institution Alberch Vié, Jordi Canela Campos, Enric Isidre Ceña Callejo, Valentín Cuadrado Pastor, Antonio del Río Fernández, José Antonio Fariñas Gómez, Isabel Fernández Ruiz, Javier Fuentes Rodríguez, José Manuel García Verdugo, José Manuel González Castaño, José Guzmán Pastor, Manuel Iglesias Vacas, Teresa Kulisevsky Bojarski, Jaime Labandeira García, José Luis Lanciego Pérez, José Luis López Barneo, José Lucas Lozano, José Javier Mena Gómez, María Ángeles Moratalla Villalba, Rosario Naranjo Orovio, José Ramón Obeso Inchausti, José Ángel * Pérez Castillo, Ana María Pérez Tur, Jordi Rodríguez Díaz, Manuel Tolosa Sarró, Eduardo Vicario Abejón, Carlos Vila Bober, Miquel University of Barcelona University of Barcelona University of Castilla-La Mancha, Albacete Autonomous University of Madrid Catalonian Institute of Bioengineering (IBEC), Barcelona University of Valencia Complutense University of Madrid University of Extremadura, Cáceres Cavanilles Institute, University of Valencia Autonomous University of Madrid Complutense University of Madrid Institute of Biomedical Research Alberto Sols (CSIC-UAM), Madrid Santa Creu i Sant Pau Hospital, Barcelona University of Santiago de Compostela Foundation for Applied Medical Research, University of Navarra, Pamplona Andalusian Public Foundation for the Health Research Management, Institute of Biomedicine of Seville (IBIS) Center for Molecular Biology Severo Ochoa (CSIC-UAM), Madrid Ramón y Cajal University Hospital, Madrid Cajal Institute CSIC, Madrid National Center of Biotechnology, CSIC, Madrid Foundation for Applied Medical Research, University of Navarra, Pamplona Institute of Biomedical Research Alberto Sols (CSIC-UAM), Madrid Institute of Biomedicine of Valencia, CSIC, Valencia University of La Laguna, Tenerife Hospital Clinic of Barcelona Cajal Institute CSIC, Madrid Vall d Hebron, Barcelona * Hospital de Madrid Foundation (since October 2014) Program 2 is coordinated by Drs. José J. Lucas (Center for Molecular Biology, CSIC, Madrid) and Eduardo Tolosa (Clinic Hospital, Barcelona). 115

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123 Group Jordi Alberch Vié Program 2 CONTACT DETAILS Group Jordi Alberch Vié Group Jordi Alberch Departament Vié Group de Biologia Jordi Alberch Cel lular, Vié Immunologia Group Jordi Alberch i Neurociències Vié Group Jordi Alberc Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Facultat Jordi de Alberch Medicina, Vié Universitat Group Jordi de Barcelona Alberch Vié Group Jord Institut d Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) Alberch Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Phone: Jordi Alberch Vié 035 Group 258 / Jordi 37252Alberch Vié Grou Fax: Jordi Alberch Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Jordi Alberc alberch@ub.edu Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Jordi Alberch Vié Group Jord LIST OF PERSONNEL Jordi Alberch Vié Principal Investigator, LIST OF PERSONNEL Full Professor Josep M Canals Coll Associate Professor Silvia Ginés Padrós Assistant Professor Esther Pérez Navarro Assistant Professor Garikoitz Azkona Mendoza Postdoctoral researcher (RETICS) Verónica Brito Postdoctoral researcher (European Huntington s Disease Network) Raquel Martín Ibáñez Postdoctoral researcher Andrés Míguez González Postdoctoral researcher (Marie Curie) Ana Cristina Saavedra Postdoctoral researcher (Juan de la Cierva) Phil Sanders Postdoctoral researcher (CHDI) Marco Straccia Postdoctoral researcher (CHDI) Xavier Xifró Collsamata Lecturer Rafael Alcalá Vida Predoctoral researcher Elena Álvarez Periel Predoctoral researcher Marta Cherubini Predoctoral researcher Jordi Creus Muncunill Predoctoral researcher Alfonso Gerardo García Díaz Barriga Predoctoral researcher Inés Guardia Pena Predoctoral researcher Andreas Mezger Predoctoral researcher Mónica Pardo Muñoz Predoctoral researcher Mar Puigdellívol Cañadell Predoctoral researcher Sara Fernández García Predoctoral researcher Shiraz Tyebji Predoctoral researcher Laura Vidal Sancho Predoctoral researcher Núria Suelves Caballol Predoctoral researcher Andrea Comella Bolla Predoctoral researcher Aina Badia Soteras Master s student Pedro Belio Mairal Master s student Beatriz Blasco Pérez Master s student Silvia Cases Cunillera Master s student Marta García Forn Master s student David López Alcalde Master s student Luis José Marte Fernández Master s student Sara Martínez Torres Master s student Blanca Peguera Carré Master s student Georgina Bombau Martínez Technician Jordi Carrere Molina Technician Ester González Guerrero Technician Cristina Herranz Sotoca Technician Ana López Alonso Technician Mª Teresa Muñoz Alarcón Technician Unai Perpiña Martin Technician 117

124 Summary Our goal is to identify new molecular mechanisms affected by mutant huntingtin (mhtt) to develop treatments to prevent or slow the progression of Huntington s disease (HD). We have demonstrated that p75ntr levels are increased in the hippocampus of HD patients and in two transgenic mouse models. This increase contributes to cognitive deficits in HD since normalization of p75ntr levels improved memory deficits and synaptic plasticity possibly through the regulation of RhoA GTPase activity. According to these results, p75ntr over-expression in the hippocampus of wild-type animals reproduced HD learning and memory deficits (Brito et al., 2014). We have also observed that levels of several PKC (α, β and δ) are decreased in different brain regions of HD models and patients, with PKCδ as the most affected possibly by increased degradation through the proteasome. We suggest that decreased PKCδ levels could be a compensatory pro-survival mechanism as silencing of this protein protects from mhtt-induced cell death in vitro (Rue et al., 2013). In another study, we determined that treatment with an antagonist of prostaglandin receptor EP1 improves R6/1 (HD mouse model) motor and cognitive deficits. These effects could be related to the restoration of synaptic proteins and the reduction in the number of mhtt inclusions in the hippocampus and striatum. Furthermore, LTP is improved in the hippocampus of mice treated with EP1 receptor antagonist. Therefore, blocking of EP1 receptors could be a good therapy to improve both motor and cognitive deficits occurring in HD (Anglada-Huguet et al., 2014). In collaboration with other groups we have described: (1) That the RTP801 protein is a novel substrate of parkin (E3 ligase). Increased levels of this pro-apoptotic protein could be involved in the neurodegeneration caused by reduced expression or activity of parkin in some cases of Parkinson s disease (Romani-Aumedes et al., 2014); (2) The over-expression of the dopamine D3 receptors could be instrumental in the activation of D1 dopamine receptors thereby causing dyskinesia in response to treatment with L-DOPA in rats previously lesioned with 6-OHDA (model of Parkinson s disease; PD). The anti-dyskinetic effects of buspirone treatment correlate with normalization of dopamine D1 receptor signaling (Azkona et al., 2014); (3) Treatment of MPTP-lesioned monkeys with L-DOPA resulted in increased levels of dopamine D2 and D3 receptors, but only the increase in D3 receptor expression correlated with the severity of dyskinesias. Monkeys having dyskinesias in response to treatment with L-DOPA exhibited increased D1 receptor signaling as we observed increased PKA activity in the postcomissural putamen (Azkona et al., 2014); (4) Treatment with 5 -deoxy-5 -methylthioadenosine (MTA) MTA displays a wide array of neuroprotective activities against different insults in vitro. In addition, reduces neuronal death in a model of epilepsy in vivo, reduces the size of the injury caused by global, but not focal, ischemia and has no effect MPTP-induced dopaminergic neurons death. However, the combined administration of MTA and an A2A adenosine receptor antagonist was found to be neuroprotective against MPTP-induced toxicity (Moreno et al., 2014); (5) A new protocol for a rapid and efficient generation of induced neural cells from hematopoietic cells (Castaño et al., 2014). Keywords BDNF, BDNF receptors, Cdk5, Cognition, Motor control, PKA/STEP, Neurogenesis, Regenerative medicine Program 2 118

125 2014 Annual Report Jordi Alberch Vié Group Publications Moreno B, Lopez I, Fernandez-Diez B, Gottlieb M, Matute C, Sanchez-Gomez MV, et al. Differential neuroprotective effects of 5'-deoxy-5'-methylthioadenosine. PloS one. 2014;9(3):e Epub 2014 Mar 5. Castano J, Menendez P, Bruzos-Cidon C, Straccia M, Sousa A, Zabaleta L, et al. Fast and efficient neural conversion of human hematopoietic cells. Stem Cell Reports. 2014;3(6): Epub 2014 Nov 13. Brito V, Giralt A, Enriquez-Barreto L, Puigdellivol M, Suelves N, Zamora-Moratalla A, et al. Neurotrophin receptor p75(ntr) mediates Huntington's disease-associated synaptic and memory dysfunction. The Journal of clinical investigation. 2014;124(10): Epub 2014 Sep 2. Romani-Aumedes J, Canal M, Martin-Flores N, Sun X, Perez-Fernandez V, Wewering S, et al. Parkin loss of function contributes to RTP801 elevation and neurodegeneration in Parkinson's disease. Cell death & disease. 2014;5:e1364. Epub 2014 Aug 7. Azkona G, Marcilla I, Lopez de Maturana R, Sousa A, Perez-Navarro E, Luquin MR, et al. Sustained increase of PKA activity in the postcommissural putamen of dyskinetic monkeys. Molecular neurobiology. 2014;50(3): Epub 2014 Apr 5. Anglada-Huguet M, Xifro X, Giralt A, Zamora-Moratalla A, Martin ED, Alberch J. Prostaglandin E2 EP1 receptor antagonist improves motor deficits and rescues memory decline in R6/1 mouse model of Huntington's disease. Molecular neurobiology. 2014;49(2): Epub 2013 Nov 7. Rue L, Alcala-Vida R, Lopez-Soop G, Creus-Muncunill J, Alberch J, Perez-Navarro E. Early downregulation of PKCδ as a pro-survival mechanism in Huntington's disease. Neuromolecular medicine. 2014;16(1): Epub 2013 Jul 30. Azkona G, Sagarduy A, Aristieta A, Vazquez N, Zubillaga V, Ruiz-Ortega JA, et al. Buspirone antidyskinetic effect is correlated with temporal normalization of dysregulated striatal DRD1 signalling in L-DOPA-treated rats. Neuropharmacology. 2014;79(): Epub 2013 Dec 10. Marin C, Bonastre M, Mengod G, Cortes R, Giralt A, Obeso JA, et al. Early L-dopa, but not pramipexole, restores basal ganglia activity in partially 6-OHDA-lesioned rats. Neurobiology of disease. 2014;64: Research Projects Code: RD12/0019 Title: RETIC de Terapia Celular Principal Investigator: Jordi Alberch (Coordinador de red: Jose Maria Moraleda) CIBERNED s collaboration: Yes Groups CIBERNED: G301; G102; G113; G208; G105; G607 Type: National Funding Agency: Fondo de Investigaciones Sanitarias Funding: Duration: Code: PI2013_08-1 Title: La dinámica mitocondrial y mitofagia como dianas terapéuticas en las Enfermedades de Parkinson y Huntington Principal Investigator: Eduardo Soriano CIBERNED s collaboration: Yes Groups CIBERNED: G301; G207; G410; G109; G408 Type: National Funding Agency: ISCIII Funding: Duration: Code: A-6965 Title: Complete Characterization of p75 Knockdown in Q111 Mouse Principal Investigator: Silvia Ginés Groups CIBERNED: Type: International Funding Agency: CHDI Funding: Duration:

126 Code: 2009SGR Title: Group de Fisiopatología de enfermedades neurodegenerativas Principal Investigator: Jordi Alberch Groups CIBERNED: Type: National Funding Agency: AGAUR, Generalitat de Catalunya Funding: Duration: Code: PI13/01250 Title: Regulación de BDNF por STEP: Implicaciones terapéuticas en enfermedades neurológicas Principal Investigator: Esther Pérez Groups CIBERNED: Type: International Funding Agency: Fondo de Investigaciones Sanitarias. Instituto de Salud Carlos III. Ministerio de Ciencia e Innovación Funding: Duration: 2014 Code: SAF Title: Regulación de la plasticidad sináptica y la disfunción neuronal por nuevos metodos de liberación de BDNF en modelos de enfermedad de Huntington Principal Investigator: Jordi Alberch Groups CIBERNED: Type: National Funding Agency: Ministerio de Ciencia e Innovación Funding: Duration: PhD Dissertations Author: Shiraz Tyebji Title: Regulation of the PKA pathway as a therapy to improve both cognitive and motor deficits in Huntington's disease Date: 29th October 2014 Supervisor: Esther Pérez Navarro Author: Mar Puigdellivol Title: Description and validation of new therapeutical targets to prevent neurodegeneration and cognitive deficits in Huntington s Disease Date: 27th November 2014 Supervisor: Silvia Ginés Padrós Program 2 120

127 Group Enric Isidre Canela Campos Program 2 CONTACT DETAILS Group Enric Isidre Canela Campos Group Enric Isidre Departamento Canela Campos de Group Bioquímica Enric y Biología Isidre Canela Molecular Campos Group Enri Isidre Canela Campos Group Enric Isidre Canela Campos Group Facultad Enric de Biología. Isidre Canela Universidad Campos de Group Barcelona Enric Isidre Canel Campos Group Enric Isidre Canela Campos Group Enric Isidre Canela Campos Group Diagonal Enric 643, Isidre planta Canela -2 Campos Grou Barcelona (Spain) Enric Isidre Canela Campos Group Enric Isidre Canela Campos Group Enric Isidre Phone: Canela Campos Group Enric Isidr Canela Campos Group Enric Isidre Canela Campos Group Enric Isidre Canela Campos Fax: Group Enric 021 Isidre 559 Canela Campo ecanela@ub.edu Group Enric Isidre Canela Campos Group Enric Isidre Canela Campos Group Enric Isidre Canela Campos Grup LIST OF PERSONNEL Enric Isidre Canela Campos Principal Investigator, Full Professor Vicent Casadó Burillo Research staff Antonio Cortés Tejedor Research staff Rafael Franco Fernández Research staff Carmen Lluís Biset Research staff Josefa Mallol Montero Research staff Gemma Navarro Brugal PhD researcher Estefanía Moreno Guillén PhD researcher Jasmina Jiménez Cano Technician Mireia Medrano Moya Predoctoral researcher David Aguinaga Andrés Predoctoral researcher Mar Rodriguez Ruiz Predoctoral researcher Verònica Casadó Anguera Predoctoral researcher Irene Reyes Resina Predoctoral researcher Edgar Angelats Canals Phd student 121

128 Summary Our area of interest has been the GPCRs in the CNS, specially their ability to form homo- and heterooligomers. We focused on discovering receptor heteromers as new targets for treatment of CNS disorders. Most relevant results in this period were: We reviewed the anatomy of the nervous system which is constituted not only by macroscopic and microscopy identifiable regions and neuronal cell types, but also by protein complexes whose identification and localization require sophisticated techniques. We also reviewed the functional and pharmacological properties of GPCR oligomers and provided some guidelines for the application of discrete direct screening and high throughput screening approaches to the discovery of receptorheteromer selective compounds. We studied the expression of heterotrimers composed of adenosine, cannabinoid and dopamine receptors (A2AR-CB1R-D2R) in the striatum of a primate model of Parkinson s disease and determined whether their expression and pharmacological properties are altered upon L-DOPA treatment. By using PLA and by identification of a biochemical fingerprint, we discovered a regional distribution of the heterotrimer that predicts differential D2R-mediated neurotransmission in primate caudateputamen. These findings constitute the first evidence of altered receptor heteromer expression in pathological conditions. We also investigated the expression of A2AR-CB1R-D2R heteromers in the striatum of both naïve and hemiparkinsonian rats (HPD-rats) bearing a unilateral 6-hydroxydopamine lesion, and assessed how receptor heteromer expression and biochemical properties were affected by L-DOPA treatment. Radioligand binding data showed that A2AR-CB1R-D2R heteromers are present in the striatum of both naïve and HPD-rats, but the heteromers are lost following acute or chronic treatment with L-DOPA. By combining BRET, bimolecular complementation techniques, and cell-signaling experiments in transfected cells, we have obtained evidences for a tetrameric stoichiometry of the D1R D3R heteromer, constituted by two interacting D1R and D3R homodimers coupled to Gs and Gi proteins, respectively. Positive cross-talk and cross-antagonism at the MAPK level were counteracted by specific synthetic peptides with amino acid sequences corresponding to D1R transmembrane domains TM5 and TM6. These results demonstrate functional selectivity of allosteric modulations within the D1R-D3R heteromer, which can be involved with the reported behavioral synergism of D1R and D3R agonists and is a potential therapeutic target for neuropsychiatric disorders. We demonstrated that different intracellular Ca2+ levels exert a differential modulation of A2AR- D2R heteromer-mediated adenylyl-cyclase and MAPK signaling in striatal cells. This depends on the ability of low and high Ca2+ levels to promote a selective interaction of the heteromer with the neuronal Ca2+-binding proteins NCS-1 and calneuron-1, respectively. These Ca2+-binding proteins differentially modulate allosteric interactions within the A2AR-D2R heteromer, which constitutes a unique cellular device that integrates extracellular (adenosine and dopamine) and intracellular (Ca2+) signals to produce a specific functional response. We showed that cocaine blockades the histamine H3R-mediated inhibition of D1R function. This blockade requires the sigma1r and occurs upon cocaine binding to sigma1-d1-h3 receptor complexes. The cocaine mediated disruption leaves an uninhibited D1R that activates Gs, freely recruits beta-arrestin, increases p-erk 1/2 levels, and induces cell death when over-activated. We showed that blockade of sigma1r by an antagonist restores the protective H3R-mediated brake on D1R signaling and prevents the cell death from elevated D1R signaling. We also checked for cannabinoid CB1 GPR55 receptor heteromers in the CNS, specifically in striatum. First, a direct interaction was demonstrated in cells transfected with the cdna of the human receptors, using BRET and PLA. A biochemical fingerprint consisting on cross-antagonism in ERK1/2 phosphorylation and GPR55-mediated NFAT activation was detected. Direct identification of Program 2 122

129 2014 Annual Report Enric Isidre Canela Campos Group GPR55 receptors in striatum was demonstrated in rat brain slices using a specific agonist. CB1-GPR55 heteromers were also identified in both caudate and putamen nuclei of a nonhuman primate. Keywords Allosteric interactions, Cocaine, G-protein Coupled receptors, L-DOPA, Proximity ligation assay, Receptor oligomerization, Sigma-1 receptor, Two-state dimer receptor model Publications Martinez-Munoz L, Barroso R, Dyrhaug SY, Navarro G, Lucas P, Soriano SF, et al. CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120iiib binding to the cell surface. Proceedings of the National Academy of Sciences of the United States of America. 2014;111(19):E Epub 2014 Apr 28. Moreno E, Moreno-Delgado D, Navarro G, Hoffmann HM, Fuentes S, Rosell-Vilar S, et al. Cocaine disrupts histamine H3 receptor modulation of dopamine D1 receptor signaling: σ1-d1-h3 receptor complexes as key targets for reducing cocaine's effects. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(10): Ugarte A, Gil-Bea F, Garcia-Barroso C, Cedazo-Minguez A, Ramirez MJ, Franco R, et al. Decreased levels of cgmp in CSF are associated with cognitive decline and amyloid pathology in Alzheimer's disease. Neuropathology and applied neurobiology. [Epub ahead of print]. Epub 2014 Dec 9. George SR, Kern A, Smith RG, Franco R. Dopamine receptor heteromeric complexes and their emerging functions. Progress in brain research. 2014;211: Franco R. Enhancing cognition before clinical symptoms of dementia. Frontiers in systems neuroscience. 2014;8:240. Epub 2014 Dec 16. Guitart X, Navarro G, Moreno E, Yano H, Cai NS, Sanchez-Soto M, et al. Functional selectivity of allosteric interactions within G protein-coupled receptor oligomers: the dopamine D1-D3 receptor heterotetramer. Molecular pharmacology. 2014;86(4): Epub 2014 Aug 5. Ferre S, Casado V, Devi LA, Filizola M, Jockers R, Lohse MJ, et al. G protein-coupled receptor oligomerization revisited: functional and pharmacological perspectives. Pharmacological reviews. 2014;66(2): Epub 2014 Feb 10. Zamarbide M, Etayo-Labiano I, Ricobaraza A, Martinez-Pinilla E, Aymerich MS, Luis Lanciego J, et al. GPR40 activation leads to CREB and ERK phosphorylation in primary cultures of neurons from the mouse CNS and in human neuroblastoma cells. Hippocampus. 2014;24(7): Epub 2014 Feb 27. Balenga NA, Martinez-Pinilla E, Kargl J, Schröder R, Peinhaupt M, Platzer W, et al. Heteromerization of GPR55 and cannabinoid CB2 receptors modulates signalling. British journal of pharmacology. 2014;171(23): Epub 2014 Sep 5. Navarro G, Aguinaga D, Moreno E, Hradsky J, Reddy PP, Cortes A, et al. Intracellular Calcium Levels Determine Differential Modulation of Allosteric Interactions within G Protein-Coupled Receptor Heteromers. Chemistry & biology. 2014;21(11): Epub 2014 Nov 6. Cortes A, Gracia E, Moreno E, Mallol J, Lluis C, Canela EI, Casado V. Moonlighting adenosine deaminase: a target protein for drug development. Medicinal research reviews. 2015;35(1): Epub 2014 Jun 16. Navarro G, Borroto-Escuela DO, Fuxe K, Franco R. Potential of caveolae in the therapy of cardiovascular and neurological diseases. Frontiers in physiology. 2014;5:370. Epub 2014 Sep 30. Franco R, Cedazo-Minguez A. Successful therapies for Alzheimer's disease: why so many in animal models and none in humans? Frontiers in pharmacology. 2014;5:146. Epub 2014 Jun 25. Fernandez-Suarez D, Celorrio M, Riezu-Boj JI, Ugarte A, Pacheco R, Gonzalez H, et al. The 123

130 monoacylglycerol lipase inhibitor JZL184 is neuroprotective and alters glial cell phenotype in the chronic MPTP mouse model. Neurobiology of aging. 2014;35(11): Epub 2014 May 28. Franco N, Franco R. Understanding the added value of g-protein-coupled receptor heteromers. Scientifica. 2014;2014: Epub 2014 Apr 22. Martinez-Pinilla E, Reyes-Resina I, Onatibia-Astibia A, Zamarbide M, Ricobaraza A, Navarro G, et al. CB1 and GPR55 receptors are co-expressed and form heteromers in rat and monkey striatum. Experimental neurology. 2014;261: Epub 2014 Jun 23. Sierra S, Luquin N, Rico AJ, Gomez-Bautista V, Roda E, Dopeso-Reyes IG, et al. Detection of cannabinoid receptors CB1 and CB2 within basal ganglia output neurons in macaques: changes following experimental parkinsonism. Brain structure & function. [Epub ahead of print]. Epub 2014 Jun 28. Brugarolas M, Navarro G, Martinez-Pinilla E, Angelats E, Casado V, Lanciego JL, et al. G-proteincoupled receptor heteromers as key players in the molecular architecture of the central nervous system. CNS neuroscience & therapeutics. 2014;20(8): Epub 2014 May 9. Cerri S, Levandis G, Ambrosi G, Montepeloso E, Antoninetti GF, Franco R, et al. Neuroprotective potential of adenosine A2A and cannabinoid CB1 receptor antagonists in an animal model of Parkinson disease. Journal of neuropathology and experimental neurology. 2014;73(5): Farre D, Munoz A, Moreno E, Reyes-Resina I, Canet-Pons J, Dopeso-Reyes IG, et al. Stronger Dopamine D1 Receptor-Mediated Neurotransmission in Dyskinesia. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Oct 26. Moreno E, Andradas C, Medrano M, Caffarel MM, Perez-Gomez E, Blasco-Benito S, et al. Targeting CB2-GPR55 receptor heteromers modulates cancer cell signaling. The Journal of biological chemistry. 2014;289(32): Epub 2014 Jun 18. Bonaventura J, Rico AJ, Moreno E, Sierra S, Sanchez M, Luquin N, et al. L-DOPA-treatment in primates disrupts the expression of A(2A) adenosine-cb(1) cannabinoid-d(2) dopamine receptor heteromers in the caudate nucleus. Neuropharmacology. 2014;79: Pinna A, Bonaventura J, Farre D, Sanchez M, Simola N, Mallol J, et al. L-DOPA disrupts adenosine A(2A)- cannabinoid CB(1)-dopamine D(2) receptor heteromer cross-talk in the striatum of hemiparkinsonian rats: biochemical and behavioral studies. Experimental neurology. 2014;253: Research Projects Code: FJL-01/01/2013 Title: Validation of mglur1/5-histamine 3 receptor heterodimers as novel targets to treat Fragile X Syndrome Principal Investigator: Peter Joseph McCormick Groups CIBERNED: Type: International Funding Agency: Fondation Jerôme Lejeune Funding: Duration: Code: AR00035 Title: Ayuda para impulsar la participación en proyectos internacionales de investigación, en el marco del Program Horizon 2012 Principal Investigator: Rafael Franco Fernández Groups CIBERNED: Type: Intramural Funding Agency: Universitat de Barcelona Funding: Duration: 2014 Code: SAF C02-01 Title: Heteromerización de receptores GPR55, CB1 y CB2: señalizacion celular, farmacología y análisis de su potencial como dianas de enfermedades neurodegenerativas Principal Investigator: Rafael Franco Fernández Groups CIBERNED: Type: International Funding Agency: Convocatoria 2012 de Research Projects Fundamental no Orientada Funding: Duration: Program 2 124

131 2014 Annual Report Enric Isidre Canela Campos Group Code: SAF Title: Heterómeros de receptores acoplados a proteína G como dianas terapéuticas de la adicción a drogas Principal Investigator: Enric Isidre Canela Campos Groups CIBERNED: Type: International Funding Agency: Ministerio de Ciencia e Innovación Funding: Duration: Code: 308/C/2013 Title: Targeting proliferation and tumor-microenvironment interactions using bifunctional compounds against cannabinoid and chemokine receptors Principal Investigator: Peter J. McCormick Groups CIBERNED: Type: Private Funding Agency: Fundació La Marató de TV3 Funding: Duration: 2014 Code: 2014-SGR-1236 Title: Neurobiología Molecular Principal Investigator: Enric I. Canela Campos Groups CIBERNED: Tipo: CCAA Funding Agency: Generalitat de Catalunya: Programs: Ajuts a Grups de recerca consolidats Funding: Duration: PhD Dissertations Author: Daniel Farré Pérez Title: Heterómeros de receptores de dopamina como dianas terapéuticas en la enfermedad de Parkinson y en discinesias inducidas por el tratamiento con L-DOPA Date: 12th December 2014 Supervisor: Vicente Casadó Burillo 125

132 Group Valentín Ceña Callejo Program 2 CONTACT DETAILS oup Valentín Ceña Callejo Group Valentín Ceña Callejo Group Valentín Unidad Ceña Callejo Asociada Group Neurodeath Valentín Ceña Callejo ña Callejo Group Valentín Ceña Callejo Group Valentín Ceña Callejo Group Universidad Valentín de Castilla-La Ceña Callejo Mancha Group Valentín a llejo Group Valentín Ceña Callejo Group Valentín Ceña Callejo Group Valentín Facultad Ceña Callejo de Medicina Group Valentín Ceña Avda. Almansa, 14 oup Valentín Ceña Callejo Group Valentín Ceña Callejo Group Valentín Ceña Callejo Group Albacete Valentín (Spain) Ceña Callejo o Valentín Ceña Callejo Group Valentín Ceña Callejo Group Valentín Ceña Callejo Phone: Group 680 Valentín a Callejo Group valentin.cena@gmail.com ña Callejo Group Valentín Ceña Callejo Group Valentín Ceña Callejo Group Valentín Ceña Callejo Group Valentín a LIST OF PERSONNEL Valentín Ceña Callejo Principal Investigator, Full Professor Inmaculada Posadas Assistant Professor Ana Belén García Sáez Lab Technician Maricarmen Mateo Gómez Lab Technician Vanesa Guijarro Lab Technician Elena Galera Lab Technician 126

133 2014 Annual Report Valentín Ceña Callejo Group Summary The research of the Neurodeath group has been focused on two different aspects: a) the use of nanoparticles, mainly dendriemrs (but also other protein-derived polymers and polyethileneimine polymers) to transfect genetic material, mainly sirna, into different cell types to knock down specific proteins involved in tumoral progression and neurodegenerative diseases and b) the role of the endoplasmic reticulum and the transcription factor HIF in the molecular mechanisms involved in neuronal death during hypoxia and neurodegenerative diseases. Related to the use of nanoparticles to remove, in a selective way, signaling proteins, it is important to note that most drug targets are proteins that are inhibited or modulated by drugs. One limitation to drug effectivity is the lack of selectivity on similar proteins (receptor types and subtypes, ionic channels, isoenzymes, etc.) producing an important number of side effects that limit treatment efficiency. An innovative therapeutic approach, that solves this limitation, is the use of sirna to remove selectively certain proteins whose function is modified in several illness which are difficult to treat because the lack of effective therapies, drug toxicity and/or the presence of resistance to treatment (cancer, TB, etc.). This approach will produce, in a short period of time, new generations of drugs, based on sirna, with the potential to revolutionize the therapeutics. Our group has focused during this year in the development of dendrimers to transfect neuronal cultures with the idea of transfecting in vivo in a near future. On the other hand, it is widely known that the endoplasmic reticulum and, specifically the PERK signaling pathway, pays a very relevant role in the neuronal death produced during the neurodegenerative diseases. Keywords Gene therapy, Dendrimers, Neurodegeneration, sirna, Transfection, Endoplasmic reticulum, PERK Publications Moreno B, Lopez I, Fernandez-Diez B, Gottlieb M, Matute C, Sanchez-Gomez MV, et al. Differential neuroprotective effects of 5'-deoxy-5'-methylthioadenosine. PloS one. 2014;9(3):e Epub 2014 Mar 5. Research Projects Code: VIHVACD Title: Desarrollo de una vacuna frente a VIH: Estudio de los cambios en la biología de células dendríticas humanas tras interacción con distintos sistemas de liberación de péptidos de VIH Principal Investigator: Mariángeles Muñoz Fernández Groups CIBERNED: Other CIBER s collaboration: Nanomedicina Type: International Funding Agency: CYTED Funding: Duration: Code: BFU C02-01 Title: Nanoparticulas no virales como vectores para sir- NA en células tumorales y neuronas Principal Investigator: Valentín Ceña Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration:

134 PhD Dissertations Author: Vanesa Ocaña Martínez Title: Papel de los canales de calcio voltaje-dependientes en el modelo de toxicidad inducido por veratridina en células cromafines bovinas Date: 16th July 2014 Supervisor: Valentín Ceña Callejo Program 2 128

135 Group Antonio Cuadrado Pastor Program 2 CONTACT DETAILS Group Antonio Cuadrado Pastor Departamento Group Antonio de Bioquímica Cuadrado e Pastor Instituto Group de Investigaciones Antonio Cuadrado Biomédicas Pastor Group Antoni Cuadrado Pastor Group Antonio Cuadrado Pastor Facultad Group de Antonio Medicina, Cuadrado Universidad Pastor Autónoma Group de Antonio Madrid Cuadrado Pasto Group Antonio Cuadrado Pastor Group Antonio Cuadrado Pastor Group Antonio Arzobispo Cuadrado Morcillo, Pastor 4 Group Antoni Madrid (Spain) Cuadrado Pastor Group Antonio Cuadrado Pastor Group Antonio Cuadrado Phone: Pastor Group Antonio Cuadrado Pasto Group Antonio Cuadrado Pastor Group Antonio Cuadrado Pastor Group Antonio Fax: Cuadrado Pastor 401Group Antoni antonio.cuadrado@uam.es Cuadrado Pastor Group Antonio Cuadrado Pastor Group Antonio Cuadrado Pastor Group Antonio Cuadrado Pastor LIST OF PERSONNEL Antonio Cuadrado Pastor Principal Investigator, UAM Isabel Lastres Becker Ramón y Cajal researcher, UAM Ana Isabel Rojo Sanchís PhD researcher, CIBERNED Ángel Juan García Yagüe PhD researcher, CIBERNED Patricia Rada Llano PhD researcher, CIBERNED Natalia Robledinos Anton Predoctoral researcher, CIBERNED Marta Pajares Cabetas FPU Predoctoral researcher, UAM Ruth Blanco García Predoctoral researcher, UAM Natalia Jiménez Moreno Master s student Joahnna Troya Valseca Student 129

136 Summary Oxidative and inflammatory stress and proteinopathy are key elements in the pathology of several neurodegenerative diseases, including Parkinson s disease (PD) and Alzheimer s disease (AD). Our group is studying the regulation of the cellular defense represented by the transcription factor NRF2 (nuclear erythroid-derived 2-like 2 factor) and one of its target genes encoding HO-1 (heme oxygenase-1). During this year, we have analyzed the protective role of NRF2 against cerebral degeneration at three levels: Characterization of the connection between GSK-3 and NRF2 in response to WNT signaling. Signaling by members of the WNT family is central to the acquisition and maintenance of dopaminergic neuronal differentiation. We have observed that NRF2 is part of a protein complex assembled with Axin1 that leads to its degradation by the GSK-3/β-TrCP similar to the β-catenin pathway. Treatment with WNT-3A breaks this complex and releases NRF2 to exert its transcriptional function. These observations have been very important for obtaining an international project CoEN (Centres of Excellence in Neurodegeneration) that we coordinate. Development of a new mouse model of tauopathy bsed on stareotaxic delivery in the hippocampus of an adeno-associated vector expressing human TAU protein (P301L) (raav6-taup301l). An advantage of this model is that it can be combined with different transgenic backgrounds, in our case the deficiency in NRF2. We have described a communication between damaged neurons and microglia for repairing the brain parenchyma that is based on the release of the chemokine Fractalkina by the neuron and the activation of a signaling pathway leading to microglial activation of NRF2. The main results of the study in mice have been validated in postmortem samples from patients with AD and progressive supranuclear palsy. We continued with our studies in a new mouse model of PD based on the expression of human α-synuclein combined with a deficiency in NRF2 expression. We are collaborating with biopharmaceutical companies to validate new compounds that activate NRF2 in preclinical models of Parkinson s disease. Keywords Oxidative stress, Neuroinflammation, Proteinopathy, Neuroprotection, NRF2, WNT, GSK-3, TAU Publications Freitas AE, Egea J, Buendia I, Navarro E, Rada P, Cuadrado A, et al. Agmatine Induces Nrf2 and Protects Against Corticosterone Effects in Hippocampal Neuronal Cell Line. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Aug 2. Ibanez L, Ferrandiz ML, Brines R, Guede D, Cuadrado A, Alcaraz MJ. Effects of Nrf2 deficiency on bone microarchitecture in an experimental model of osteoporosis. Oxidative medicine and cellular longevity. 2014;2014: Epub 2014 Jul 7. Delgado-Buenrostro NL, Medina-Reyes EI, Lastres-Becker I, Freyre-Fonseca V, Ji Z, Hernandez-Pando R, et al. Nrf2 protects the lung against inflammation induced by titanium dioxide nanoparticles: A positive regulator role of Nrf2 on cytokine release. Environmental toxicology. [Epub ahead of print]. Epub 2014 Feb 26. Rojo AI, McBean G, Cindric M, Egea J, Lopez MG, Rada P, et al. Redox control of microglial function: molecular mechanisms and functional significance. Antioxidants & redox signaling. 2014;21(12): Epub 2014 May 5. Program 2 130

137 2014 Annual Report Antonio Cuadrado Pastor Group Egea J, Buendia I, Parada E, Navarro E, Rada P, Cuadrado A, et al. The melatonin-sulforaphane hybrid ITH12674 affords neuroprotection in oxidative stress conditions by a "Drug - Prodrug" mechanism of action. British journal of pharmacology. [Epub ahead of print]. Epub 2014 Nov 26. Rojo AI, Rada P, Mendiola M, Ortega-Molina A, Wojdyla K, Rogowska-Wrzesinska A, et al. The PTEN/ NRF2 axis promotes human carcinogenesis. Antioxidants & redox signaling. 2014;21(18): Epub 2014 Jul 31. Cuadrado A, Martin-Moldes Z, Ye J, Lastres-Becker I. Transcription factors NRF2 and NF-κB are coordinated effectors of the Rho family, GTP-binding protein RAC1 during inflammation. The Journal of biological chemistry. 2014;289(22): Epub 2014 Apr 22. Rada P, Rojo AI, Offergeld A, Feng GJ, Velasco-Martin JP, Gonzalez-Sancho JM, et al. WNT-3A Regulates an Axin1/NRF2 Complex That Regulates Antioxidant Metabolism in Hepatocytes. Antioxidants & redox signaling. [Epub ahead of print]. Epub 2014 Oct 21. Simon D, Herva ME, Benitez MJ, Garrido JJ, Rojo AI, Cuadrado A, et al. Dysfunction of the PI3K- Akt-GSK-3 pathway is a common feature in cell culture and in vivo models of prion disease. Neuropathology and applied neurobiology. 2014;40(3): Lastres-Becker I, Innamorato NG, Jaworski T, Rabano A, Kügler S, Van Leuven F, et al. Fractalkine activates NRF2/NFE2L2 and heme oxygenase 1 to restrain tauopathy-induced microgliosis. Brain: a journal of neurology. 2014;137(1): Research Projects Code: Fundación Salud Title: Caracterización del sistema de degradación del proteasoma y autofagia en un modelo AAV6-a-sinucleína de la enfermedad de Parkinson: implicación del factor de transcripción Nrf2 Principal Investigator: Isabel Lastres Becker Groups CIBERNED: Type: National Funding Agency: Fundación Salud 2000 Funding: Duration: 2014 Code: Biogen Idec 2013 Title: Targeting Nrf2 with BG 12 to modify Parkinson s disease progression in the AAV6 α synuclein mouse model Principal Investigator: Antonio Cuadrado Pastor Groups CIBERNED: Type: Private Funding Agency: Biogen Idec Funding: Duration: 2014 Code: COEN2008 Title: WNT signaling: biomarker and target evaluation in Alzheimer s disease Principal Investigator: Antonio Cuadrado Pastor Groups CIBERNED: Type: International Funding Agency: Instituto de Salud Carlos III Funding: Duration: 2014 Code: SAF R Title: Papel de NRF2 como modulador antioxidante de la neuroinflamación en la enfermedad de Alzheimer Principal Investigator: Antonio Cuadrado Pastor Groups CIBERNED: Type: International Funding Agency: MINECO Funding: Duration:

138 Group José Antonio del Río Fernández Program 2 CONTACT DETAILS oup José Antonio del Río Fernández Group José Antonio del Molecular Río Fernández and Group Cellular José Neurobiotechnology Antonio del Río Fernández Group osé Antonio del Río Fernández Group José Antonio del Río Fernández Institute for Group Bioengineering José Antonio of Catalonia del Río Fernández (IBEC) Group José tonio del Río Fernández Group José Antonio del Río Fernández Group José Antonio del Baldiri Río Fernández Reixac, Group José Antonio Barcelona (Spain) l Río Fernández Group José Antonio del Río Fernández Group José Antonio Phone: del +34 Río 934 Fernández Group / José Antonio del o Fernández Group José Antonio del Río Fernández Group jadelrio@ub.edu José Antonio del / jadelrio@ibecbarcelona.eu Río Fernández Group José Antonio del tonio del Río Fernández Group José Antonio del Río Fernández Group José Antonio del Río Fernández Group José Antonio LIST OF PERSONNEL José Antonio del Río Fernández Principal Investigator Rosalina Gavín Marín Senior researcher Vanessa Gil Fernández Postdoctoral Fellow Ariadna Pérez Balaguer Postdoctoral Fellow Cristina Vergara Paños PhD student Andreu Matamoros Anglès PhD student Ágata Mata Rodríguez PhD student Diego Reginensi Espinosa PhD student Laura Urrea Zazurca PhD student Miriam Segura Feliu Lab Technician Anna Prieto Colomina Student Lorena Sureiro Ballesteros Student Sara Nocentini PhD student Patricia Carulla Martí PhD student Oscar Seira Oriach Postdoctoral Fellow 132

139 2014 Annual Report José Antonio del Río Fernández Group Summary Our research interests are focused on four main aspects of Developmental Neurobiology and Neurodegeneration: Analysis of cell migration and functions of Cajal-Retzius cells during cortical development. Cajal- Retzius (CR) cells are transient pioneer neurons that migrate from specific regions of the developing brain (e.g., the cortical hem) to cover the complete neocortex. Functions of these cells are associated to the correct formation of cell layer in developing cortex. Altered migration of Cajal-Retzius cells impairs further development of the neocortex leading to an inverted cortex. The migration of these cells is controlled by a cytokine CxCL12 (expressed in meningeal cells) and its receptor CxCR4 (expressed in CR cells). We have determined that a secreted semaphorin (Sema3E) expressed by radial glia is sable to modulate the migration of CR cells by acting on its receptor PlexinD1. This signalling, modulate CxCL12/CxCR4 signalling in CR cells leading to a decreased migration that could help CR cells to distribute in the developing cortex. Absence of this signalling lead to impaired CR cell migration generating a misfunctional cortex. These data was published in Nature Communications. Cell therapy and pharmacological treatment to potentiate axon regeneration in lesioned central nervous system. In the last years we focused to genetically modify olfactory ensheathing cells (OECs) to be able to increase their survival and migration in the lesioned spinal cord. Thus, results were recently published in Cellular and Molecular Life Science. In the experiments OECs were genetically modified to overexpress the ectodomain of Nogo Receptor being able to increase their migration in inhibitory substrates in vitro and in contused spinal cord in vivo. This study was developed in collaboration of X. Navarro and F. Wandosell groups of CIBERNED, and X. Trepat and J. Samitier from CIBERBBN. Neurodegenerative diseases. We determined the role of a natural neural cellular prion protein protein (PrPc) in the evolution of the Alzheimer s disease (published in Molecular Neurobiology). Results point to PrPc as key neuroprotective factor in Alzheimer s disease. In fact, when present Tau phosphorylation by ADDLs is impaired. However in Braak stages III, PrPc down-regulates by unknown reasons and Tau phosphorylation increases. These experiments were developed in vitro, in mouse models of AD as well in postmortem human samples. Experiments were developed in collaboration with I. Ferrer and F. Wandosell from CIDERNED. Next experiments will continue in this line but also will expand to Parkinson s disease. Our hypothesis is that PrPc is a cross-link protein between different neurodegenerative diseases presenting taupathy. In addition, our group collaborates in some omics studies of specific markers in rapid progressive dementia (e.g., CJD) in collaboration with I. Ferrer. Development of new lab on a chip devices for neurobiological research. We recently developed a new device able to reproduce the neuromuscular junction in a single chip (published in RSC advances). In this device, cultured motoneurons are able to establish the neuromuscular junction of concrete and specific foam. Thus current experiments of our group in collaboration with groups of IBEC and CIBERBBN aimed to develop new lab on chip devices to mimics and modulate particular neurobiological processes. For example: axonal lesions that mimic Wallerian-like degeneration, molecular gradient generation for migrating neurons and in silico 3D modelization, or neurodegenerative diseases: Parkinson s disease). 133

140 Keywords Cell proliferation, PrPc-dependent genomic signature, Beta-amyloid, mirna Publications Tong Z, Seira O, Casas C, Reginensi D, Homs-Corbera A, Samitier J, et al. Engineering a functional neuro-muscular junction model in a chip. RSC ADVANCES. 2014;4: Epub Gil V, Nocentini S, Del Rio JA. Historical first descriptions of Cajal-Retzius cells: from pioneer studies to current knowledge. Frontiers in neuroanatomy. 2014;8:32. Epub 2014 May 27. Vergara C, Ordonez-Gutierrez L, Wandosell F, Ferrer I, Del Rio JA, Gavin R. Role of PrPC Expression in Tau Protein Levels and Phosphorylation in Alzheimer's Disease Evolution. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Jun 26. Bribian A, Nocentini S, Llorens F, Gil V, Mire E, Reginensi D, et al. Sema3E/PlexinD1 regulates the migration of hem-derived Cajal-Retzius cells in developing cerebral cortex. Nature communications. 2014;5:4265. Epub 2014 Jun 27. Llorens F, Zafar S, Ansoleaga B, Shafiq M, Blanco R, Carmona M, et al. Subtype and regional regulation of prion biomarkers in sporadic Creutzfeldt-Jakob disease. Neuropathology and applied neurobiology. [Epub ahead of print]. Epub 2014 Aug 18. Seira O, Del Rio JA. Glycogen synthase kinase 3 beta (GSK3β) at the tip of neuronal development and regeneration. Molecular neurobiology. 2014;49(2): Llorens F, Ferrer I, del Rio JA. Gene expression resulting from PrPC ablation and PrPC overexpression in murine and cellular models. Molecular neurobiology. 2014;49(1): Research Projects Code: PI11/03028 Title: DEMTEST: Biomarker based diagnosis of rapid progressive dementias Principal Investigator: Pascual Sánchez CIBERNED s collaboration: Yes Groups CIBERNED: G601; G114; G503; G609 Type: Others Funding Agency: Instituto de Salud Carlos III - Comunidad Económica Europea Funding: Duration: Code: BIO12/AL/004 Title: Estudio de la replicación tipo prión en modelos celulares obtenidos mediante la reprogramción de células somáticas de pacientes geneticamente susceptibles a la enfermedad de Alzheimer Principal Investigator: Joaquín Castilla Groups CIBERNED: Type: Private Funding Agency: Fundación Vasca de Innovación e Investigación Sanitarias Funding: Duration: Code: SRI-NEUROMEMs Title: Neuromems Principal Investigator: José Antonio del Río CIBERNED s collaboration: Groups CIBERNED: Type: Private Funding Agency: Obra Social La Caixa Funding: Duration: Code: BFU Program 2 134

141 2014 Annual Report José Antonio del Río Fernández Group Title: Nuevas funciones de PLEXIND1/SEMA3E, PRPC y las proteínas asociadas a la mielina durante el desarrollo de la corteza cerebral de roedores y en neurodegeneración Principal Investigator: José Antonio del Río Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: Code: AC50 Title: Protecting the food chain of prions: Shaping European priorities through basic and applied research Principal Investigator: Jesús Requena (USC) Groups CIBERNED: Type: International Funding Agency: FP/ - KBBE Funding: Duration: PhD Dissertations Author: Sara Nocentini Title: Potential of genetically modified ensheathing cells for regeneration after spinal cord injury Date: 7th March 2014 Supervisor: José Antonio del Río Fernández 135

142 Group Isabel Fariñas Gómez Program 2 CONTACT DETAILS oup Isabel Fariñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Departamento Gómez Group de Biología Isabel Celular Fariñas Gómez Group bel Fariñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Gómez Group Universidad Isabel Fariñas de Valencia Gómez Group Isabel riñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Doctor Gómez Moliner, Group 50 Isabel Fariñas Burjassot (Spain) oup Isabel Fariñas Gómez Group Isabel Fariñas Gómez Phone: Group +34 Isabel Fariñas 784 Gómez (despacho/office) Group Isabel 3246 Fariñas (lab) Gómez Group bel Fariñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Gómez Group Fax: Isabel +34 Fariñas Gómez 404 Group Isabel Isabel.farinas@uv.es riñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas Gómez Group Isabel Fariñas LIST OF PERSONNEL Isabel Fariñas Principal Investigator, Full Professor Cell Biology Francisco Pérez Sánchez Scientific staff Martina Kirstein Scientific staff Sacramento Rodríguez Ferrón Scientific staff Ana Pérez Villalba Postdoctoral Fellow Eva Porlan Alonso Postdoctoral Fellow José Manuel Morante Postdoctoral Fellow Salomé Sirerol Piquer Postdoctoral Fellow Alex Bizy Postdoctoral Fellow María Pimentel Santillana Postdoctoral Fellow Carmen Ovejero Benito Postdoctoral Fellow Beatriz Martí Prado PhD student Raúl Soriano Cantón PhD student Germán Belenguer Sánchez PhD student Ana Domingo Muelas PhD student Raquel Montalbán PhD student María José Palop Benlloch Technician Fabrice Durupt Technician Elba Barberà Ferragud Technician 136

143 2014 Annual Report Isabel Fariñas Gómez Group Summary Our laboratory studies two topics related to neurodegeneration and cell therapy: 1) The study of cellular and molecular alterations that underlie degeneration of dopaminergic neurons, and 2) The study of neural stem cell (NSC) self-renewal. In recent years, we have also been working in combining both aspects by studying the regulation of adult neurogenesis in parkinsonism and during aging. During 2014, we have unveiled two aspects of the behavior of NSCs and their relationship to the architecture of the subependymal zone. On one hand, we have shown that adhesive interactions mediated by N-cadherin between NSCs and other cells in their niche regulate NSC quiescence and have identified MT5-MMP as the metalloproteinase responsible for the proteolytic cleavage of the ectodomain of the N-cadherin that is necessary for NSC activation. Furthermore, we have shown that the neurotrophin NT-3, which was best known for its effects on neuronal survival, is an endothelium-derived quiescence-promoting factor for adult NSCs through effects on the induction of nitric oxide production by the NSCs themselves. This is a novel function for a neurotrophin and NT-3 becomes the first soluble factor inducing the quiescence that is necessary for the long-term maintenance of the NSCs. Our results indicate that the NSCs can regulate their quiescence in response to molecules secreted by endothelia and present in the extracellular medium of the capillaries that irrigate the area as well as in cerebrospinal fluid. In 2014, we have also worked with several groups of CIBERNED in three different topics. On one hand, we have collaborated with the groups of Drs. Vila and Obeso in demonstrating that pathological forms of alpha-synuclein present in Parkinson s disease patients are able to initiate the neurodegenerative process in normal brains of mice and monkeys. Furthermore, in collaboration with Drs. Toledo and Moratalla, we have discovered that dopamine not only controls the proliferation of the progenitors of the subependymal zone but also regulates the potential status of NSCs and their maintenance over time and that alpha-synuclein, present in the dopaminergic terminals that innervate the subependymal zone, is necessary for the action of dopamine. We have also observed an interaction between neurogenesis and aging in mouse models of accelerated senescence and the effect of alpha-synuclein on dopaminergic neurodegeneration in such genetic background. Another line that we have continued to explore this year, in collaboration with the group of Dr. Cuadrado and in the context of one of the intramural collaborative projects already completed, has been the role of the transcription factor Nrf2 in the behavior of NSCs. Finally, in the context of a new intramural collaborative project with the teams led by Drs. Fernández Chacon and Lucas Lozano, we have been analyzing adult neurogenesis in mice deficient in the synaptic regulator CSP-alpha. Keywords Adult neurogenesis and cell therapy, Neural stem cells, Neurogenic niches, Neurotrophic factors, Mouse models of neurodegenerative disease, Parkinson s disease 137

144 Publications Delgado AC, Ferron SR, Vicente D, Porlan E, Perez-Villalba A, Trujillo CM, et al. Endothelial NT-3 delivered by vasculature and CSF promotes quiescence of subependymal neural stem cells through nitric oxide induction. Neuron. 2014;83: Epub 18 julio Porlan E, Marti-Prado B, Morante-Redolat JM, Consiglio A, Delgado AC, Kypta R, et al. MT5-MMP regulates adult neural stem cell functional quiescence through the cleavage of N-cadherin. Nature cell biology. 2014;16(7): Epub 2014 Jun 22. Recasens A, Dehay B, Bove J, Carballo-Carbajal I, Dovero S, Perez A, et al. Lewy body extracts from parkinson's disease brains trigger α-synuclein pathology and neurodegeneration in mice and monkeys. Annals of neurology Mar;75(3): Epub 2014 Feb 18. Research Projects Code: Convenio Botín-UV Title: Estudio de células madre en el ámbito de las investigaciones básicas en terapia celular Principal Investigator: Isabel Fariñas Groups CIBERNED: Type: National Funding Agency: Fundación Botín-Banco Santander Funding: Duration: Code: RD12/0019/0008 Title: RETIC de Terapia Celular Principal Investigator: Isabel Fariñas (Coordinador de red: Jose María Moraleda) CIBERNED s collaboration: Yes Groups CIBERNED: G301; G102; G113; G208; G105; G607 Type: National Funding Agency: Fondo de Investigaciones Sanitarias Funding: Duration: Code: PROMETEOII/2013/020 Title: Efectos del microambiente vascular en las células madre del cerebro adulto Principal Investigator: Isabel Fariñas Groups CIBERNED: Type: International Funding Agency: Conselleria de Educacion de la Generalitat Valenciana Funding: Duration: Program 2 138

145 Group Javier Fernández Ruiz Program 2 CONTACT DETAILS Group Javier Fernández Ruiz Group Javier Fernández Departamento Ruiz Group de Javier Bioquímica Fernández y Biología Ruiz Group Molecular Javier IIIFernández Ruiz Javier Fernández Ruiz Group Javier Fernández Ruiz Group Facultad Javier Fernández Medicina, Ruiz Universidad Group Javier Complutense Fernández Ruiz Group Fernández Ruiz Group Javier Fernández Ruiz Group Javier Pabellón Fernández IV 4ª Ruiz Planta, Group Laboratorios Javier Fernández 9, 11 y 12 Ruiz Group Javie Avenida Complutense s/n Group Javier Fernández Ruiz Group Javier Fernández Ruiz Group Javier Fernández Ruiz Group Madrid (Spain) Javier Fernández Ruiz Javier Fernández Ruiz Group Javier Fernández Ruiz Group Javier Fernández Ruiz Phone: Group +34 Javier Fernández 450 Ruiz Group Fax: Fernández Ruiz Group Javier Fernández Ruiz Group Javier jjfr@med.ucm.es Fernández Ruiz / J.Fernandez-Ruiz@ciberned.es Group Javier Fernández Ruiz Group Javie LIST OF PERSONNEL Javier Fernández Ruiz Principal Investigator, Full Professor José Antonio Ramos Atance Full Professor Mariluz Hernández Gálvez Associate Professor María Gómez Ruiz Assistant Professor Eva de Lago Femia Assistant Professor Onintza Sagredo Ezkioga Assistant Professor Concepción García García Postdoctoral researcher Mª Ruth Pazos Rodríguez Postdoctoral researcher Carmen Rodríguez Cueto PhD student Cristina Palomo Garo PhD student María Gómez Cañas PhD student Sara Valdeolivas Rojas PhD student Francisco Espejo Porras PhD student María Ceprián Costoso PhD student Yolanda García Movellán Administrative Assistant Moisés García-Arencibia Postdoctoral researcher Miguel Moreno Martet PhD student 139

146 Summary The work during 2014 has been concentrated in the study of the neuroprotective potential of cannabinoids in several chronic neurodegenerative disorders with motor symptoms, and, in particular, in the identification of cellular and molecular mechanisms that underlie these effects. In the case of Huntington s disease, the work has continued the collaboration with the companies GW Pharmaceuticals and Vivacell Biotechnology-Spain in relation with the evaluation of the neuroprotective effects of different phytocannabinoids (D9-THC, cannabidiol, cannabigerol) in experimental models of this disease, in particular R6/2 mice. At the same time, we have initiated some studies addressed to determine the molecular and cellular mechanisms underlying these effects using in vitro models of this disease. Lastly, we are preparing for publication the clinical trial conducted in the Hospital Ramón y Cajal in Madrid in patients using Sativex, trial in which our group has participated in the analysis of different biomarkers in collaboration with other CIBERNED groups. In the case of Parkinson s disease, the work has also continued the collaboration with the two companies, in the case of GW Pharmaceuticals in investigating the advantages of the combination of cannabidiol and D9-THCV for the symptomatic and neuroprotective treatment of this disease, and, in the case of Vivacell Biotechnology, in determining the potential of cannabigerol-quinones (they are ligands of PPARs) and cannabidiol-quinones (CB2 and PPAR agonists) in experimental models of this disease. We are particularly interested in the role of CB2 receptors in relation with the control of inflammation in this disease, by comparing different experimental models and including studies in mice deficient for this receptor and also LRRK2 mutant mice, a genetic model of Parkinson s disease. This study is being conducted within a project funded by the Michael J. Fox Foundation. In the case of amyotrophic lateral sclerosis, the work has again addressed the identification of the changes that the disease provokes in the endocannabinoid system and completed the evaluation of phytocannabinoids and derivatives of the companies GW Pharmaceuticals and Vivacell Biotechnology, using SOD-1 mutant mice and also TDP-43 transgenic mice, which also serves for the experimental study of frontotemporal dementia. In the case of Vivacell Biotechnology, these studies are being carried out within a project funded by the last call of the RETOS-COLABORACION program of the MINECO. We have also continued the biochemical and pharmacological (with Sativex) studies initiated in 2013 in dogs with degenerative myelopathy, an ALS-like disorder also produced by SOD-1 mutations. Lastly, in the case of spinocerebellar ataxias, the work has been concentrated in finishing the biochemical and histopathological studies addressed to determine the changes that the disease produces in the endocannabinoid system (ligands, receptors and enzymes) in the cerebellum of SCA-3 transgenic mice, and we have initiated the first pharmacological studies with different cannabinoids in these mice, studies addressed to evaluate the neuroprotective potential of these compounds. Keywords Cannabinoids, CB1 receptors, CB2 receptors, Neuroprotection, Huntington s disease, Parkinson s disease, Amyotrophic lateral sclerosis, Spinocerebellar ataxias Program 2 140

147 2014 Annual Report Javier Fernández Ruiz Group Publications Rojo AI, McBean G, Cindric M, Egea J, Lopez MG, Rada P, et al. Redox control of microglial function: molecular mechanisms and functional significance. Antioxidants & redox signaling. 2014;21(12): Epub 2014 May 5. Rojo AI, Rada P, Mendiola M, Ortega-Molina A, Wojdyla K, Rogowska-Wrzesinska A, et al. The PTEN/ NRF2 axis promotes human carcinogenesis. Antioxidants & redox signaling. 2014;21(18): Epub 2014 Jul 31. Rada P, Rojo AI, Offergeld A, Feng GJ, Velasco-Martin JP, Gonzalez-Sancho JM, et al. WNT-3A Regulates an Axin1/NRF2 Complex That Regulates Antioxidant Metabolism in Hepatocytes. Antioxidants & redox signaling. [Epub ahead of print]. Epub 2014 Oct 21. Chiarlone A, Bellocchio L, Blazquez C, Resel E, Soria-Gomez E, Cannich A, et al. A restricted population of CB1 cannabinoid receptors with neuroprotective activity. Proceedings of the National Academy of Sciences of the United States of America. 2014;111(22): Epub 2014 May 19. Devinsky O, Cilio MR, Cross H, Fernandez-Ruiz J, French J, Hill C, et al. Cannabidiol: pharmacology and potential therapeutic role in epilepsy and other neuropsychiatric disorders. Epilepsia. 2014;55(6): Epub 2014 May 22. Fernandez-Ruiz J, Hernandez ML, Garcia-Movellan Y. Cannabinoids and the brain: new hopes for new therapies. In: Di Marzo V, Ed. Cannabinoids. Wiley-Blacwell Publications, USA; p Rodriguez-Cueto C, Benito C, Fernandez-Ruiz J, Romero J, Hernandez-Galvez M, Gomez-Ruiz M. Changes in CB(1) and CB(2) receptors in the post-mortem cerebellum of humans affected by spinocerebellar ataxias. British journal of pharmacology. 2014;171(6): Moreno-Martet M, Espejo-Porras F, Fernandez-Ruiz J, de Lago E. Changes in endocannabinoid receptors and enzymes in the spinal cord of SOD1(G93A) transgenic mice and evaluation of a Sativex( ) -like combination of phytocannabinoids: interest for future therapies in amyotrophic lateral sclerosis. CNS neuroscience & therapeutics. 2014;20(9): Epub 2014 Apr 7. Fernandez-Fernandez C, Callado LF, Giron R, Sanchez E, Erdozain AM, Lopez-Moreno JA, et al. Combining rimonabant and fentanyl in a single entity: preparation and pharmacological results. Drug design, development and therapy. 2014;8: Epub 2014 Feb 20. Ooms M, Rietjens R, Rangarajan JR, Vunckx K, Valdeolivas S, Maes F, et al. Early decrease of type 1 cannabinoid receptor binding and phosphodiesterase 10A activity in vivo in R6/2 Huntington mice. Neurobiology of aging. 2014;35(12): Epub 2014 Jun 16. Rodriguez-Cueto C, Benito C, Romero J, Hernandez-Galvez M, Gomez-Ruiz M, Fernandez-Ruiz J. Endocannabinoid-hydrolysing enzymes in the post-mortem cerebellum of humans affected by hereditary autosomal dominant ataxias. Pathobiology: journal of immunopathology, molecular and cellular biology. 2014;81(3): Epub 2014 Mar 14. Garcia MC, Cinquina V, Palomo-Garo C, Rabano A, Fernandez-Ruiz J. Identification of CB2 receptors in human nigral neurons that degenerate in Parkinson's disease. Neuroscience letters. 2015;587:1-4. Epub 2014 Dec 3. Fernandez-Ruiz J, de Lago E, Gomez-Ruiz M, Garcia C, Sagredo O, Garcia-Arencibia M. Neurodegenerative disorders other than multiple sclerosis. In: Pertwee R, Ed. Handbook of Cannabis. School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, UK; Valdeolivas S, Navarrete C, Cantarero I, Bellido ML, Munoz E, Sagredo O. Neuroprotective Properties of Cannabigerol in Huntington's Disease: Studies in R6/2 Mice and 3-Nitropropionate-lesioned Mice. Neurotherapeutics: the journal of the American Society for Experimental NeuroTherapeutics. [Epub ahead of print]. Epub 2014 Sep 25. Moreno E, Andradas C, Medrano M, Caffarel MM, Perez-Gomez E, Blasco-Benito S, et al. Targeting CB2-GPR55 receptor heteromers modulates cancer cell signaling. The Journal of biological chemistry. 2014;289(32): Epub 2014 Jun 18. Erdozain AM, Rubio M, Valdizan EM, Pazos A, Meana JJ, Fernandez-Ruiz J, et al. The endocannabinoid system is altered in the post-mortem prefrontal cortex of alcoholic subjects. Addiction biology. [Epub 141

148 ahead of print]. Epub 2014 Jul 8. Perez M, Cuadros R, Pallas-Bazarra N, Garcia C, Langa E, Jurado-Arjona J, et al. Boronate-tau mediated uptake in neurons. Journal of Alzheimer's disease: JAD. 2014;40(1): Gonzalez-Naranjo P, Perez-Macias N, Campillo NE, Perez C, Aran VJ, Giron R, et al. Cannabinoid agonists showing BuChE inhibition as potential therapeutic agents for Alzheimer's disease. European journal of medicinal chemistry. 2014;73: Research Projects Code: CIBERNED 2013/05 Title: Identificación y caracterización molecular de subpoblaciones de receptores cannabinoides en poliglutaminopatías Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G201; G303; G304; G305 Type: International Funding Agency: CIBERNED Funding: Duration: Code: S2010/BMD-2308 Title: Neurofarmacología del sistema endocannabinoide: del laboratorio a la clínica Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G303; G304; G305 Other CIBER s collaboration: CIBERSAM Tipo: CCAA Funding Agency: Comunidad de Madrid Funding: Duration: Code: MJFOXF Title: Cannabinoid CB2 receptors as a new target for the treatment of disease progression in Parkinson s disease: studies in LRRK2-transgenic mice Principal Investigator: Javier Fernández Ruiz / Mª Concepción García García Groups CIBERNED: Type: International Funding Agency: Michael J. Fox Foundation Funding: N.D. Duration: Code: SAF Title: El sistema cannabinoide como diana para una terapia neuroprotectora en la esclerosis lateral amiotrófica Principal Investigator: Javier Fernández Ruiz Groups CIBERNED: Type: International Funding Agency: Ministerio de Economía y Competitividad. Dirección Gral. de Investigación y Gestión Plan Nacional I+D+i Funding: Duration: Code: FIS PIE13/00040 Title: Modulation of cellular micro RNAs as a therapeutic strategy for he cure of HIV infection Principal Investigator: Santiago Moreno Groups CIBERNED: Type: International Funding Agency: ISCIII. Estrategia de Salud Proyectos Integrados de Excelencia en los Institutos de Investigación Acreditados Funding: N.D. Duration: Code: GR3/14 Title: Program de Financiación Groups de Investigación. Cannabinoides Principal Investigator: Javier Fernández Ruiz / Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G303; G305 Type: Intramural Funding Agency: UCM - Banco Santander Funding: Duration: Code: ALZH (ELF) Title: Role of the endocannabinoid system in TDP43-related dementia Principal Investigator: Eva de Lago Femia Program 2 142

149 2014 Annual Report Javier Fernández Ruiz Group Groups CIBERNED: Type: International Funding Agency: Alzheimer s Association Funding: Duration: Code: GW2013 Title: Studies with phytocannabinoids as disease modifying agents in different neurodegenerative disorders Principal Investigator: Javier Fernández Ruiz Groups CIBERNED: Type: Private Funding Agency: GW Pharmaceuticals, Ltd Funding: Duration: PhD Dissertations Author: Miguel Moreno Martet Title: Cannabinoids as disease modifiers in Multiple Sclerosis and Amyotrophic Lateral Sclerosis Date: 7th July 2014 Supervisors: Javier Fernández Ruiz / Eva de Lago Femia 143

150 Group José Manuel Fuentes Rodríguez Program 2 CONTACT DETAILS oup José Manuel Fuentes Rodríguez Group José Manuel Dpto. Fuentes Bioquímica Rodríguez y Biología Group Molecular José Manuel y Genética Fuentes Rodríguez po sé Manuel Fuentes Rodríguez Group José Manuel Fuentes Rodríguez Group José F. Manuel Enfermería Fuentes y T.O. Rodríguez Group Universidad de Extremadura ntes Rodríguez Group José Manuel Fuentes Rodríguez Group José Manuel Fuentes Rodríguez Group José Manuel Avenida de la Universidad s/n oup José Manuel Fuentes Rodríguez Group José Manuel Fuentes Rodríguez Group José Cáceres Manuel (Spain) Fuentes Rodríguez po sé Manuel Fuentes Rodríguez Group José Manuel Fuentes Rodríguez Phone: Group José 257 Manuel 450 (ext Fuentes 51286) Rodríguez Group Fax: ntes Rodríguez Group José Manuel Fuentes Rodríguez Group José Manuel Fuentes jfuentes@unex.es Rodríguez Group José Manuel oup José Manuel Fuentes Rodríguez Group José Manuel Fuentes Rodríguez Group José Manuel Fuentes Rodríguez po LIST OF PERSONNEL José Manuel Fuentes Rodríguez Principal Investigator, Full Professor Rosa Ana González Polo Postdoctoral Fellow Mireia Niso Santano Postdoctoral Fellow Juan de la Cierva José Manuel Bravo San Pedro Postdoctoral Fellow Rubén Gómez Sánchez Postdoctoral Fellow Elisa Pizarro Estrella Postdoctoral student Ignacio Casado Naranjo Head of Department Sokhna Maryama Seydina Yakhine DIOP PhD student Mario Rodríguez Arribas PhD student (FPU program) Guadalupe Martínez Chacon Technician CIBERNED María Pura Delgado Luceño Technician 144

151 2014 Annual Report José Manuel Fuentes Rodríguez Group Summary Our efforts during 2014 have continued focusing on the study of basic molecular mechanisms involved in the pathogenesis of Parkinson s disease, autophagy regulation and role of PARK genes derived proteins in both processes. Moreover we have also continued our collaboration and service delivery to Extremadura Health Service in molecular diagnosis of Parkinson s disease. In 2014 we have continued to study the role of PINK1 in mitochondrial dynamics. Thus we have tested as PINK1 deletion of autophagy and mitophagy promotes an effective flow evidenced by the use of constructs with LC3 and p62- autophagy markers and double labeled with fluorophore GFP and Cherry. This increase in autophagy was reversed by using not only the wild form of PINK1 but mutant K219M which has abolished its kinase activity, E240K and L489P (both affecting the kinase domain and related to Parkinson s disease) and the double mutant E240K and L489P. Therefore we have shown that the control of autophagy levels by PINK1 are independent of its kinase domain. Therefore we will continue our work in this line to show how PINK1 controls cell homeostasis by regulating this cellular mechanism. Following the work we carried out since 2009 in collaboration with the group of Dr. Adolfo López de Munain (CIBERNED), have continued to characterize the basal macroautophagy deregulation in fibroblasts from Parkinson patients carriers of pathogenic G2019S mutation in LRRK2. Fibroblasts from individuals with G2019S LRRK2 mutation presents levels of autophagy basally higher than those observed in fibroblasts from individuals without the mutation, this flare is accompanied by an increased sensitivity of the cells to toxic substances, such as MPP+, regarding Parkinson s disease. After a molecular dissection we found that the MEK/ERK pathway could play an active role in the deregulation of basal autophagy and consequent increased sensitivity presented in individuals with LRRK2 G2019S mutation. Finally we are moving in functional studies using fibroblasts from individuals with the mutation of LRRK2 R1441G. In this case we have also seen an accelerated basal autophagy, however no higher degradation than fibroblasts from individuals without this mutation. In both models we begin the analysis of the expression of actyltransferase and deacetylase enzymes and the degree of acetylation of proteins related to autophagy, since this posttranslational marking is showing as a key regulator in macroautophagic response Finally we have followed interested in the study of general mechanisms of regulation of autophagy, this part of our business is still carried out in collaboration with the laboratory of Dr. Guido Kroemer of Apoptosis, Cancer & Immunity Laboratory, INSERM Cordeliers Research Center, Paris, France. Regarding our collaboration with the Extremadura Health Service should be noted that we have signed four agreements to provide services in relation to the molecular diagnosis of Parkinson s. It has been increased to twenty the number of products in our portfolio of services, and participate in research training for healthcare professionals. Keywords Pesticides and Parkinson s Disease, Roles of apoptosis and autophagy in Parkinson s Disease, Detection of mutations in PARK genes, Functional roles of PARK gene-derived proteins (synuclein, parkin, DJ-1, PINK1 and LRRK2), Nrf2/keap1 axis 145

152 Publications Michaud M, Xie X, Bravo-San Pedro JM, Zitvogel L, White E, Kroemer G. An autophagy-dependent anticancer immune response determines the efficacy of melanoma chemotherapy. Oncoimmunology. 2014;3(7):e Epub 2014 Jul 3. Casado Naranjo I, Portilla Cuenca JC, Duque de San Juan B, Garcia AF, Sevilla RR, Serrano Cabrera A, et al. Association of Vascular Factors and Amnestic Mild Cognitive Impairment: A Comprehensive Approach. Journal of Alzheimer's disease: JAD. 2015;44(2): Epub 2014 Oct 31. Sistigu A, Yamazaki T, Vacchelli E, Chaba K, Enot DP, Adam J, et al. Cancer cell-autonomous contribution of type I interferon signaling to the efficacy of chemotherapy. Nature medicine. 2014;20(11): Epub 2014 Oct 26. Galluzzi L, Vacchelli E, Bravo-San Pedro JM, Buque A, Senovilla L, Baracco EE, et al. Classification of current anticancer immunotherapies. Oncotarget. 2014;5(24): Molinuevo JL, Casado-Naranjo I. Clinical profile of Alzheimer's disease: is the age of the patient a decisive factor? Results of the INFLUENCE study. Journal of Alzheimer's disease: JAD. 2014;39(2): Pietrocola F, Malik SA, Marino G, Vacchelli E, Senovilla L, Chaba K, et al. Coffee induces autophagy in vivo. Cell cycle (Georgetown, Tex.). 2014;13(12): Epub 2014 Apr 25. Galluzzi L, Bravo-San Pedro JM, Vitale I, Aaronson SA, Abrams JM, Adam D, et al. Essential versus accessory aspects of cell death: recommendations of the NCCD Cell death and differentiation. 2015;22(1): Epub 2014 Sep 19. Yakhine-Diop SM, Bravo-San Pedro JM, Gomez-Sanchez R, Pizarro-Estrella E, Rodriguez-Arribas M, Climent V, et al. G2019S LRRK2 mutant fibroblasts from Parkinson's disease patients show increased sensitivity to neurotoxin 1-methyl-4-phenylpyridinium dependent of autophagy. Toxicology. 2014; 324:1-9. Epub 2014 Jul 10. Gonzalez-Polo RA, Bravo-San Pedro JM, Gomez-Sanchez R, Pizarro-Estrella E, Niso-Santano M, Fuentes JM. Links Between Paraquat and Parkinson s Disease. In: Kostrzewa R, Ed. Handbook of Neurotoxicity. Springer; p Bonora M, Bravo-San Pedro JM, Kroemer G, Galluzzi L, Pinton P. Novel insights into the mitochondrial permeability transition. Cell cycle (Georgetown, Tex.). 2014;13(17): Gomez-Suaga P, Fdez E, Fernandez B, Martinez-Salvador M, Blanca Ramirez M, Madero-Perez J, et al. Novel insights into the neurobiology underlying LRRK2-linked Parkinson's disease. Neuropharmacology. 2014;85: Epub 2014 May 24. Galluzzi L, Bravo-San Pedro JM, Kroemer G. Organelle-specific initiation of cell death. Nature cell biology. 2014;16(8): Marino G, Pietrocola F, Eisenberg T, Kong Y, Malik SA, Andryushkova A, et al. Regulation of autophagy by cytosolic acetyl-coenzyme A. Molecular cell. 2014;53(5): Epub 2014 Feb 20. Marino G, Niso-Santano M, Baehrecke EH, Kroemer G. Self-consumption: the interplay of autophagy and apoptosis. Nature reviews. Molecular cell biology. 2014;15(2): Epub 2014 Jan 8. Pietrocola F, Lachkar S, Enot DP, Niso-Santano M, Bravo-San Pedro JM, Sica V, et al. Spermidine induces autophagy by inhibiting the acetyltransferase EP300. Cell death and differentiation. 2015;22(3): Epub 2014 Dec 19. Gomez-Sanchez R, Gegg ME, Bravo-San Pedro JM, Niso-Santano M, Alvarez-Erviti L, Pizarro-Estrella E, et al. Mitochondrial impairment increases FL-PINK1 levels by calcium-dependent gene expression. Neurobiology of disease. 2014;62: Program 2 146

153 2014 Annual Report José Manuel Fuentes Rodríguez Group Research Projects Code: PI12/02280 Title: Acetilación, autofagia y Enfermedad de Parkinson Principal Investigator: José Manuel Fuentes Rodríguez Groups CIBERNED: Type: International Funding Agency: Ministerio de Economía y Competitividad. Instituto de Salud Carlos III. Fondo de Investigación Sanitaria Funding: Duration: Code: GR10054 Title: Ayudas para la consolidación y apoyo a los Groups de investigacion inscritos en el Catálogo de Groups de Investigación de Extremadura Principal Investigator: José Manuel Fuentes Rodríguez Groups CIBERNED: Tipo: CCAA Funding Agency: Consejería Infraestructura y Desarrollo Tecnológico de la Junta de Extremadura Funding: Duration: Code: PI11/00040 Title: Estudio de la importancia de la autofagia en células procedentes de enfermos de Parkinson con mutaciones en genes park Principal Investigator: Rosa Ana González Polo Groups CIBERNED: Type: National Funding Agency: FIS. ISCIII. MICINN Funding: Duration: PhD Dissertations Author: Elisa Pizarro Estrella Title: El factor de transcripción Nrf2 como nueva diana terapéutica para la enfermedad de parkinson Date: 18th July 2014 Supervisor: José Manuel Fuentes Rodríguez Author: Ignacio Casado Naranjo Title: Análisis de marcadores de inflamación y arterioesclerosis carotidea subclínica en la relación entre el síndrome metabólico y el deterioro cognitivo ligero Date: 10th October 2014 Supervisor: José Manuel Fuentes Rodríguez 147

154 Group José Manuel García Verdugo Program 2 CONTACT DETAILS oup José Manuel García Verdugo Group José Manuel García Laboratorio Verdugo de Group Neurobiología José Manuel Comparada García Verdugo (SS7) Group José anuel García Verdugo Group José Manuel García Instituto Verdugo Cavanilles Group de José Biodiversidad Manuel García y Biología Verdugo Evolutiva Group José Manuel rcía Verdugo Group José Manuel García Verdugo Group José Manuel García Universidad Verdugo Group de Valencia José Manuel García Catedrático José Beltrán, 2 rdugo Group José Manuel García Verdugo Group José Manuel García Verdugo Group Paterna José Manuel (Spain) García Verdugo oup José Manuel García Verdugo Group José Manuel García Verdugo Group Phone: José +34 Manuel García 587Verdugo Group Fax: José Manuel García Verdugo Group José Manuel García Verdugo Group José Manuel j.manuel.garcia@uv.es García Verdugo Group José anuel García Verdugo Group José Manuel García Verdugo Group José Manuel García Verdugo Group José Man LIST OF PERSONNEL José Manuel García Verdugo Principal Investigator, Full Professor Juan Antonio Barcia Albacar Assistant researcher, Neurosurgeon José Miguel Soria López Assistant researcher María Salomé Sirerol Piquer Postdoctoral researcher Vicente Herranz Pérez Postdoctoral researcher CIBERNED Sara García Gil-Perotín Postdoctoral Fellow Arantxa Cebrián Silla PhD student María Durán Moreno PhD student Susana González Granero Technician CIBERNED 148

155 2014 Annual Report José Manuel García Verdugo Group Summary Our group remains focused mainly on the research of stem cells in the adult mammalian central nervous system. These cells not only generate new neurons, but also glial cells, among which, we are focused primarily on the generation of new oligodendroglial cells. Besides the better understanding that provides the basic research on the dynamics of these cells, we are expanding our work towards various diseases that affect the nervous system, both studying what happens at the cellular level and analyzing different therapeutic methods. Finally, we are offering an extensive microscopy platform, which combines light and electron microscopy (and they correlation) in a variety of tissues and species, which is rendering us a broader picture of other systems. During 2014 we completed several studies at the level of basic research. We analyzed the organization of the spinal cord central canal of mammals, conducting a morphological and molecular study of its cellular organization, both in human and non-human primates. We also advanced in our knowledge of the factors that are required for specificity and differentiation of tanycytes of the third ventricle of mammals. Moreover, we concluded an interesting study about the functionality of axons (originating from a small group of neurons in the raphe) present on most of the ventricular walls of mice, revealing synaptic contacts on neural stem cells and ependymal cells. These contacts may play an important role in controlling neurogenesis. Furthermore, in the SVZ we completed two additional studies: one concerning the high ability of this region to regenerate myelin following a process of demyelination after being subjected to radiation, and another based on the analysis of the changes that occur with aging on astrocytes and ependymal cells. Moreover, we also conducted an analysis of the mechanism of cell signaling by which neural stem cells communicate with their micro-environment through exosomes, specifically by analyzing their ability for transferring interferon gamma after being subjected to an inflammation process, allowing some degree of interaction with the immune system. And finally, we have participated in the analysis of stem cells of the carotid body, finding that control of their proliferation is determined by the secretion of endothelin-1 via vesicles (similarly to synaptic contacts) from the glomus cells. On the other hand, we have also participated in the study of different pathologies related to the nervous system. We have established a relationship between alterations affecting the cells that compose the ventricles and various diseases. For example, we have observed cytoarchitectonic changes that occur in different animal models of hydrocephalus. In one of them, fractone presence is associated with alterations in the cellular organization of the ventricular walls in an occlusive model of hydrocephalus. While in another, we have related the Dvl protein family, present in the environment of the basal bodies of cilia, with the establishment of the planar polarity of ependymal cells. As a result, mutation of these proteins causes circulation deficits of the cerebrospinal fluid. Also related to ciliopathies, we collaborated in the identification of the gene whose mutations produce a type of oral-facial-digital syndrome, since they affect the correct elongation of centrioles. And finally, we could establish in a different study, a possible correlation between the number and structure of the cilia of ependymal cells of the SVZ and the degree of development of ependymomas. Keywords Neurogenesis, Stem cells, Subventricular zone, Third ventricle, Oligodendrocytes 149

156 Publications Capilla-Gonzalez V, Cebrian-Silla A, Guerrero-Cazares H, Garcia-Verdugo JM, Quinones-Hinojosa A. Age-related changes in astrocytic and ependymal cells of the subventricular zone. Glia. 2014;62(5): Garcia-Gonzalez D, Murcia-Belmonte V, Esteban PF, Ortega F, Diaz D, Sanchez-Vera I, et al. Anosmin-1 over-expression increases adult neurogenesis in the subventricular zone and neuroblast migration to the olfactory bulb. Brain structure & function. [Epub ahead of print]. Epub 2014 Oct 10. Flores-Bellver M, Bonet-Ponce L, Barcia JM, Garcia-Verdugo JM, Martinez-Gil N, Saez-Atienzar S, et al. Autophagy and mitochondrial alterations in human retinal pigment epithelial cells induced by ethanol: implications of 4-hydroxy-nonenal. Cell death & disease. 2014;5:e1328. Epub 2014 Jul 17. Tong CK, Chen J, Cebrian-Silla A, Mirzadeh Z, Obernier K, Guinto CD, et al. Axonal control of the adult neural stem cell niche. Cell stem cell. 2014;14(4): Epub 2014 Feb 20. Cossetti C, Iraci N, Mercer TR, Leonardi T, Alpi E, Drago D, et al. Extracellular vesicles from neural stem cells transfer IFN-γ via Ifngr1 to activate Stat1 signaling in target cells. Molecular cell. 2014;56(2): Epub 2014 Sep 18. Campos-Ordonez T, Herranz-Perez V, Chaichana KL, Rincon-Torroella J, Rigamonti D, Garcia-Verdugo JM, et al. Long-term hydrocephalus alters the cytoarchitecture of the adult subventricular zone. Experimental neurology. 2014;261: Epub 2014 May 21. Ohata S, Nakatani J, Herranz-Perez V, Cheng J, Belinson H, Inubushi T, et al. Loss of Dishevelleds disrupts planar polarity in ependymal motile cilia and results in hydrocephalus. Neuron. 2014;83(3): Epub 2014 Jul 18. Jones J, Estirado A, Redondo C, Pacheco-Torres J, Sirerol-Piquer MS, Garcia-Verdugo JM, et al. Mesenchymal stem cells improve motor functions and decrease neurodegeneration in ataxic mice. Molecular therapy: the journal of the American Society of Gene Therapy. 2015;23(1): Epub 2014 Jul 29. Bobadilla M, Sainz N, Rodriguez JA, Abizanda G, Orbe J, de Martino A, et al. MMP-10 is required for efficient muscle regeneration in mouse models of injury and muscular dystrophy. Stem cells (Dayton, Ohio). 2014;32(2): Iglesias-Garcia O, Baumgartner S, Macri-Pellizzeri L, Rodriguez-Madoz JR, Abizanda G, Guruceaga E, et al. Neuregulin-1β Induces Mature Ventricular Cardiac Differentiation from Induced Pluripotent Stem Cells Contributing to Cardiac Tissue Repair. Stem cells and development. [Epub ahead of print]. Epub 2014 Oct 20. Paradells S, Rocamonde B, Llinares C, Herranz-Perez V, Jimenez M, Garcia-Verdugo JM, et al. Neurotoxic effects of ochratoxin A on the subventricular zone of adult mouse brain. Journal of applied toxicology: JAT. [Epub ahead of print]. Epub 2014 Sep 25. Valencia M, Caparros-Martin JA, Sirerol-Piquer MS, Garcia-Verdugo JM, Martinez-Glez V, Lapunzina P, et al. Report of a newly indentified patient with mutations in BMP1 and underlying pathogenetic aspects. American journal of medical genetics. Part A. 2014;164A(5): Epub 2014 Mar 19. Ballester-Lurbe B, Gonzalez-Granero S, Mocholi E, Poch E, Garcia-Manzanares M, Dierssen M, et al. RhoE deficiency alters postnatal subventricular zone development and the number of calbindinexpressing neurons in the olfactory bulb of mouse. Brain structure & function. [Epub ahead of print]. Epub 2014 Jul 10. Gomez-Nicola D, Suzzi S, Vargas-Caballero M, Fransen NL, Al-Malki H, Cebrian-Silla A, et al. Temporal dynamics of hippocampal neurogenesis in chronic neurodegeneration. Brain: a journal of neurology. 2014;137(Pt 8): Epub 2014 Jun 18. Alfaro-Cervello C, Cebrian-Silla A, Soriano-Navarro M, Garcia-Tarraga P, Matias-Guiu J, Gomez-Pinedo U, et al. The adult macaque spinal cord central canal zone contains proliferative cells and closely resembles the human. The Journal of comparative neurology. 2014;522(8): Salvatierra J, Lee DA, Zibetti C, Duran-Moreno M, Yoo S, Newman EA, et al. The LIM Homeodomain Program 2 150

157 2014 Annual Report José Manuel García Verdugo Group Factor Lhx2 Is Required for Hypothalamic Tanycyte Specification and Differentiation. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(50): Thauvin-Robinet C, Lee JS, Lopez E, Herranz-Perez V, Shida T, Franco B, et al. The oral-facial-digital syndrome gene C2CD3 encodes a positive regulator of centriole elongation. Nature genetics. 2014;46(8): Epub 2014 Jul 6. Capilla-Gonzalez V, Guerrero-Cazares H, Bonsu JM, Gonzalez-Perez O, Achanta P, Wong J, et al. The subventricular zone is able to respond to a demyelinating lesion after localized radiation. Stem cells (Dayton, Ohio). 2014;32(1): Garcia-Contreras M, Vera-Donoso CD, Hernandez-Andreu JM, Garcia-Verdugo JM, Oltra E. Therapeutic potential of human adipose-derived stem cells (ADSCs) from cancer patients: a pilot study. PloS one. 2014;9(11):e Epub 2014 Nov 20. Alfaro-Cervello C, Soriano-Navarro M, Ramirez M, Bernet L, Martinez Banaclocha M, Cano R, et al. Ultrastructural Pathology of Anaplastic and Grade II Ependymomas reveals Distinctive Ciliary Structures - Electron Microscopy Redux. Ultrastructural pathology. 2014;39(1):23-9. Epub 2014 May 15. Platero-Luengo A, Gonzalez-Granero S, Duran R, Diaz-Castro B, Piruat JI, Garcia-Verdugo JM, et al. An O2-sensitive glomus cell-stem cell synapse induces carotid body growth in chronic hypoxia. Cell. 2014; 156(1-2): Garcia-Parra P, Naldaiz-Gastesi N, Maroto M, Padin JF, Goicoechea M, Aiastui A, et al. Murine muscle engineered from dermal precursors: an in vitro model for skeletal muscle generation, degeneration, and fatty infiltration. Tissue engineering. Part C, Methods. 2014;20(1): Arana M, Gavira JJ, Pena E, Gonzalez A, Abizanda G, Cilla M, et al. Epicardial delivery of collagen patches with adipose-derived stem cells in rat and minipig models of chronic myocardial infarction. Biomaterials. 2014;35(1): Research Projects Code: RD12/0019/0016 Title: Red Terapia Celular: NEUROCEL-ELA. Nodo Valencia Principal Investigator: Jose Manuel García Verdugo CIBERNED s collaboration: Yes Groups CIBERNED: G301; G113; G105; G208; G102; G607 Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: SAF Title: Análisis del proceso de mielinización en el estriado: efectos del EGF E IGF-1 Principal Investigator: Jose Manuel García Verdugo Groups CIBERNED: Type: National Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: PRI-PIMNEU Title: Biomaterials scaffoldings for brain reconstruction in stroke. ERA-NET NEURON Principal Investigator: Juan Antonio Barcia Albacar Groups CIBERNED: Type: International Funding Agency: Ministerio de Ciencia e Innovación Funding: Duration: Code: PROMETEOII/2014/075 Title: Células madre adultas: Aplicaciones en terapia regenerativa Principal Investigator: Jose Manuel García Verdugo Groups CIBERNED: Tipo: CCAA Funding Agency: Conselleria de Cultura, Educación y Deportes, Generalitat Valenciana Funding: Duration: Code: CB06/05/1131 Title: CIBERNED Principal Investigator: Jose Manuel García Verdugo 151

158 Groups CIBERNED: Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: 479/C/2012 Title: Efecto de la migración de células madre mediante campos magnéticos en la recuperación del infarto cerebral Principal Investigator: Juan Sahuquillo Barris Groups CIBERNED: Tipo: CCAA Funding Agency: Fundació la Marató de TV3 Funding: Duration: Code: Title: Multipotent Adult Progenitor Cells to treat Stroke II (Strokemap II) Principal Investigator: Catherine Verfaillie Groups CIBERNED: Type: European Funding Agency: European Comission Funding: N.D. Duration: Code: RD06/0010/0022 Title: Red de Terapia Celular. Nodo Valencia Principal Investigator: Jose Manuel García Verdugo Groups CIBERNED: Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: SEP Title: Use of ACA-generated stem cells for cure of neural degenerative disease like Cerebral Ischemia Principal Investigator: Zorica Becker Kojic Groups CIBERNED: Type: European Funding Agency: Unión Europea - H2020 Funding: N.D. Duration: 2014 Program 2 152

159 Group José González Castaño Program 2 CONTACT DETAILS Group José González Departamento Castaño Group de Bioquímica José González e Instituto Castaño de Group Investigaciones José González Biomédicas Castaño Alberto Group José Sols González Castañ Group José González Castaño Group José González Castaño Group José González UAM-CSIC. Castaño Lab Group B13 José Gonzále Castaño Group José González Castaño Group José González Castaño Group Facultad José de González Medicina Castaño UAM Group Jos Arzobispo Morcillo s/n González Castaño Group José González Castaño Group José González Castaño Group José Madrid González (Spain) Castaño Group José González Castaño Group José González Castaño Group José González Castaño Phone: Group José 975 González 427 Castaño Grou Fax: José González Castaño Group José González Castaño Group José González Castaño joseg.castano@uam.es Group José González Castaño LIST OF PERSONNEL José González Castaño Principal Investigator Raúl Sánchez Lanzas Predoctoral researcher Teresa Bermejo Pastor Research Techician 153

160 Summary Proteostasis is essential for cell homeostasis and function. It includes the mechanisms for keeping a healthy cell proteome: protein synthesis, folding, protein-protein interactions, post-translational modifications (PTMs) and degradation. Alpha-synuclein is a small protein implicated in the pathophysiology of Parkinson s disease (PD). We have shown that alpha-synuclein interacts with the C8 (α7) subunit of the proteasome through its N-terminal region and directly degraded. Aggregated alpha-synucleins (wild type and the point mutants A30P, E46K and A53T) is also degraded by the proteasome with a reduced rate. This reduction is not due to simple aggregation or fibril formation, but to oxidation of the N-terminal Met residues of alpha-synuclein. The inhibition was reversed by treatment of the oxidized protein with methionine sulfoxide reductases (MsrA plus MsrB). Similarly, treatment with H2O2 of N2A cells expressing alpha-synuclein resulted in the inhibition of its degradation and is also reverted by co-transfection of MsrA plus MsrB. As a consequence, oxidative stress, a common feature of PD and other synucleinopathies, promotes a RedOx change in the proteostasis of alphasynuclein due to Met oxidation and reduced proteasomal degradation; compromised reversion of those oxidative changes would result in the accumulation of PTM (oxidative damaged) alpha-synuclein likely contributing to the pathogenesis of PD. Exogenous addition of aggregated alpha-synuclein has been shown to promote aggregation of cell endogenous alpha-synuclein and then propagated to nearby cells. Antibodies against alpha-synuclein counteract that saltatory transmission of aggregation. Circulating anti-snca antibodies has been described in PD patients and healthy controls, but they have been poorly characterized. We have assessed the prevalence of alpha-synuclein reactivity in human subjects carrying the LRRK2 mutation, idiopathic PD (ipd) patients, and healthy controls. Clear positive reactivity showed no correlation with age, sex, years of evolution, or the disability scores for PD patients and reactivity was not prevalent in human patients with other neurological or autoimmune diseases. Thirteen of the positive individuals were carriers of LRRK2 mutations, three positive were ipd patients, and five positive were healthy controls. Epitope mapping showed that antibodies against the N-terminal (a.a. 1-60) or C-terminal (a.a ) regions of alpha-synuclein predominate in LRRK2 mutation carriers and ipd patients, being N122 a key determinant for recognition by the anti-c-terminal directed antibodies. These positive individuals cluster within families carrying the LRRK2 mutation indicating possible genetic or common environmental factors in the generation of anti-snca antibodies. We were unable to correlate seropositivity with any clinical parameter or disease evolution of PD. These results suggest that further studies of family cohorts of PD patients and healthy controls should be undertaken, to progress in the understanding of the possible relationship of anti-snca antibodies and PD pathology. Those studies may help define a possible immunotherapy approach for treatment of PD patients. Our final contribution from last year was a review on the mammalian proteins shown to directly interact with specific 20S proteasomal subunits and those subjected to ubiquitin-independent proteasomal degradation that include alpha-synuclein and DJ-1 L166P, implicated in PD. We propose that systematic studies of the specificity of the interactions, the elucidation of the protein regions implicated in the interactions (that may or may not be followed by degradation) and competition experiments between proteins known to interact with the same proteasomal subunit, are needed. Those studies should provide a coherent picture of the molecular mechanisms governing the interactions of cellular proteins with proteasomal subunits, and their relevance to cell proteostasis and cell functioning, another key to understand neurodegenerative disease and other proteinopathies. Keywords Synuclein, RedOx, Proteasome, Human antibodies, Proteolysis, Proteostasis, Protein aggregation, Parkinson Program 2 154

161 2014 Annual Report José González Castaño Group Publications Sanchez-Lanzas R, Castano JG. Proteins directly interacting with mammalian 20S proteasomal subunits and ubiquitin-independent proteasomal degradation. Biomolecules. 2014;4(4): Epub 2014 Dec 19. Alvarez-Castelao B, Gorostidi A, Ruiz-Martinez J, Lopez de Munain A, Castano JG. Epitope Mapping of Antibodies to Alpha-Synuclein in LRRK2 Mutation Carriers, Idiopathic Parkinson Disease Patients, and Healthy Controls. Frontiers in aging neuroscience. 2014;6:169. Epub 2014 Jul 15. Alvarez-Castelao B, Goethals M, Vandekerckhove J, Castano JG. Mechanism of cleavage of alphasynuclein by the 20S proteasome and modulation of its degradation by the RedOx state of the N-terminal methionines. Biochimica et biophysica acta. 2014;1843(2): Research Projects Code: S2010_BMD Title: Redes de señalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas Principal Investigator: José González Castaño CIBERNED s collaboration: Yes Groups CIBERNED: G401; G104; G111; G307; G409 Other CIBER s collaboration: Yes (CIBERER) Type: International Funding Agency: Comunidad de Madrid Funding: Duration: Code: BM1307 Title: 2013 Cost Action Proteostasis Principal Investigator: Manuel S. Rodríguez Rosa Barrios Groups CIBERNED: Type: International Funding Agency: UE Funding: 500 Duration:

162 Group Manuel Guzmán Pastor Program 2 CONTACT DETAILS oup Manuel Guzmán Pastor Group Manuel Guzmán Departamento Pastor Group de Bioquímica Manuel Guzmán y Biología Pastor Molecular Group Manuel I Guzmán stor Group Manuel Guzmán Pastor Group Manuel Guzmán Facultad Pastor de Biología, Group Manuel Universidad Guzmán Complutense Pastor Group Manuel zmán Pastor Group Manuel Guzmán Pastor Group Manuel Guzmán Pastor Group Manuel Madrid Guzmán (Spain) Pastor Group Phone: anuel Guzmán LIST OF Pastor PERSONNEL Group Manuel Guzmán Pastor Group Manuel Guzmán Pastor Group Manuel Guzmán Pastor Fax: oup Manuel Guzmán Pastor Group Manuel Guzmán Pastor Group Manuel Guzmán mguzman@quim.ucm.es Pastor Group Manuel Guzmán Guzmán Pastor Group Manuel Guzmán Pastor Group Manuel Guzmán Pastor Group Manuel Guzmán Pastor Group LIST OF PERSONNEL Manuel Guzmán Pastor Principal Investigator, Full Professor Ismael Galve-Roperh Principal Co-Investigator, Associate Professor Guillermo Velasco Díez Associate Professor Cristina Blázquez Ortiz Associate Professor Luigi Bellocchio Postdoctoral Fellow Javier Díaz Alonso Postdoctoral Fellow Anna Paola Chiarlone Postdoctoral Fellow Raquel Bajo Graneras Postdoctoral Fellow Adán de Salas Quiroga PhD student Andrea Ruiz Calvo PhD student Eva Resel Mariné Lab Manager Elena García Taboada Lab Technician 156

163 2014 Annual Report Manuel Guzmán Pastor Group Summary Our research focuses globally on the study of the molecular and cellular mechanisms underlying the control of neural cell physiopathology by the endocannabinoid system. Thus, in 2014 we have kept on studying how this system tunes neural progenitor proliferation, differentiation and survival. Specifically, in the context of our CIBERNED program, we have evaluated the role of the endocannabinoid system in Huntington s disease. As a matter of fact, Huntington s disease constitutes, in our opinion, the best currently available model disease to assess the pathophysiological relevance and therapeutic potential of the endocannabinoid system in neurodegenerative diseases. This is due to several reasons: (1) CB1 is the most abundant G protein-coupled receptor in the brain; specifically, it is highly expressed in the basal ganglia at synapses established by neurons containing GABA [especially medium-sized spiny neurons (MSNs), the cells that primarily degenerate in Huntington s disease] or glutamate (especially corticostriatal projecting neurons, which critically control MSN function) as transmitters, and play a key role in the control of motor behaviour (one of the processes that is most characteristically affected in Huntington s disease). (2) A remarkable and dorsolaterally-selective down-regulation of the CB1 receptor has been documented in the basal ganglia of Huntington s disease patients and animal models, and, of interest, this loss of CB1 receptors seems to reflect, at least in part, the neuron-damage pattern characteristic of the disease. (3) This loss of CB1 receptors occurs at early stages of Huntington s disease and prior to the appearance of overt clinical symptoms, neurodegeneration and bulk changes in other neurochemical parameters. In 2014, our studies on the endocannabinoid system in Huntington s disease have aimed at defining (1) whether stimulation of the endocannabinoid system in Huntington s disease models promotes neuroprotection and therefore delays the onset and/or attenuates the progression of the pathology, (2) whether the alterations of CB1 cannabinoid receptors observed in MSNs and/or corticostriatal terminals at early stages of the disease are involved in the pathogenesis of Huntington s disease, and (3) whether the induction of CB2 cannabinoid receptors in reactive microglial cells modulates the excitotoxicity processes associated with Huntington s disease. We aim at unravelling the molecular and cellular bases as well as the potential clinical relevance of those processes. Keywords Endocannabinoid system, Huntington s disease, Neuroprotection, Neurogenesis, Cell signalling, Experimental therapeutics Publications Chiarlone A, Bellocchio L, Blazquez C, Resel E, Soria-Gomez E, Cannich A, et al. A restricted population of CB1 cannabinoid receptors with neuroprotective activity. Proceedings of the National Academy of Sciences of the United States of America. 2014;111(22): Epub 2014 May 19. Moreno E, Andradas C, Medrano M, Caffarel MM, Perez-Gomez E, Blasco-Benito S, et al. Targeting CB2-GPR55 receptor heteromers modulates cancer cell signaling. The Journal of biological chemistry. 2014; 289(32): Epub 2014 Jun 18. Diaz-Alonso J, Aguado T, de Salas-Quiroga A, Ortega Z, Guzman M, Galve-Roperh I. CB1 Cannabinoid Receptor-Dependent Activation of mtorc1/pax6 Signaling Drives Tbr2 Expression and Basal Progenitor Expansion in the Developing Mouse Cortex. Cerebral cortex (New York, N.Y.: 1991). [Epub ahead of print]. Epub 2014 Mar 7. Pietropaolo S, Bellocchio L, Ruiz-Calvo A, Cabanas M, Du Z, Guzman M, et al. Chronic cannabinoid receptor stimulation selectively prevents motor impairments in a mouse model of Huntington's 157

164 disease. Neuropharmacology. 2015;89: Epub 2014 Aug 11. Oddi D, Subashi E, Middei S, Bellocchio L, Lemaire-Mayo V, Guzman M, et al. Early Social Enrichment Rescues Adult Behavioral and Brain Abnormalities in a Mouse Model of Fragile X Syndrome. Neuropsychopharmacology: official publication of the American College of Neuropsychopharmacology. [Epub ahead of print]. Epub 2014 Oct 28. Salazar M, Lorente M, Garcia-Taboada E, Perez Gomez E, Davila D, Zuniga-Garcia P, et al. Loss of Tribbles pseudokinase-3 promotes Akt-driven tumorigenesis via FOXO inactivation. Cell death and differentiation. 2015;22(1): Epub 2014 Aug 29. Maccarrone M, Guzman M, Mackie K, Doherty P, Harkany T. Programming of neural cells by (endo) cannabinoids: from physiological rules to emerging therapies. Nature reviews. Neuroscience. 2014;15(12): Soria-Gomez E, Bellocchio L, Reguero L, Lepousez G, Martin C, Bendahmane M, et al. The endocannabinoid system controls food intake via olfactory processes. Nature neuroscience. 2014;17(3): Epub 2014 Feb 9. Research Projects Code: GR3/14 Title: Program de Financiación Groups de Investigación. Cannabinoides Principal Investigator: Javier Fernández Ruiz, Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G303; G305 Type: Intramural Funding Agency: UCM - Banco Santander Funding: Duration: Code: CIBERNED 2013/05 Title: Identificación y caracterización molecular de subpoblaciones de receptores cannabinoides en poliglutaminopatías Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G201; G303; G304; G305 Type: International Funding Agency: CIBERNED Funding: Duration: Code: S2010/BMD2336 Title: Estudio de la biología de células madre neurales para su empleo en terapia celular en enfermedades neurodegenerativas Principal Investigator: Rosario Moratalla Villalba CIBERNED s collaboration: Yes Groups CIBERNED: G204; G108; G305 Tipo: CCAA Funding Agency: Comunidad de Madrid Funding: Duration: Code: PI12/02248 Title: Mecanismos de regulación de la autofagia en células tumorales: muerte mediada por autofagia en respuesta a fármacos antitumorales y participación en el control de la tumorigénesis Principal Investigator: Guillermo Velasco Díez Groups CIBERNED: Type: National Funding Agency: MINECO-ISCIII-FIS Funding: Duration: Code: S2010/BMD-2308 Title: Neurofarmacología del sistema endocannabinoide: del laboratorio a la clínica Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G303; G304; G305 Other CIBER s collaboration: CIBERSAM Tipo: CCAA Funding Agency: Comunidad de Madrid Funding: Duration: Code: SAF Title: Neuroprotección por el receptor CB1 cannabinoide en la enfermedad de Huntington: relevancia de las vías corticoestriatales directa e indirecta Program 2 158

165 2014 Annual Report Manuel Guzmán Pastor Group Principal Investigator: Manuel Guzmán Pastor Groups CIBERNED: Type: National Funding Agency: MINECO-Plan Nacional Funding: Duration: Code: PI12/00919 Title: Papel del receptor CB1 cannabinoide en alteraciones de la neurogénesis cortical: Implicaciones en excitabibilidad neuronal y función corticoespinal Principal Investigator: Ismael Galve Roperh Groups CIBERNED: Type: National Funding Agency: MINECO-ISCIII-FIS Funding: Duration: PhD Dissertations Author: Javier Díaz Alonso Title: Papel del receptor CB1 cannabinoide en el desarrollo de la corteza cerebral Date: 19th June 2014 Supervisor: Ismael Galve Roperh Author: Anna Paola Chiarlone Title: Papel neuroprotector del receptor CB1 cannabinoide en la enfermedad de Huntington Date: 8th July 2014 Supervisor: Manuel Guzmán Pastor 159

166 Group Teresa Iglesias Vacas Program 2 CONTACT DETAILS oup Teresa Iglesias Vacas Group Teresa Iglesias Instituto Vacas Group de Investigaciones Teresa Iglesias Biomédicas Vacas Group Alberto Teresa Sols Iglesias Vacas resa Iglesias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas (CSIC-UAM) Group y CIBERNED Teresa Iglesias (ISCIII) Vacas Group esias Vacas Group Teresa Iglesias Vacas Departamento Group Teresa de Fisiopatología Iglesias Vacas Endocrina Group y Teresa del Sistema Iglesias Nervioso Vacas Group Teresa Arturo Duperier, 4 oup Teresa Iglesias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas Madrid (Spain) resa Iglesias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas Phone: Group +34 Teresa Iglesias 487 Vacas Group esias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas Group Fax: Teresa +34 Iglesias Vacas 401Group Teresa tiglesias@iib.uam.es oup Teresa Iglesias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas Group Teresa Iglesias Vacas LIST OF PERSONNEL Teresa Iglesias Vacas Principal Investigator, CSIC Tenured Scientist Celia López Menéndez Postdoctoral Fellow Lucía García Guerra Postdoctoral Fellow Ana María del Puerto del Pino Postdoctoral Fellow Andrea Gamir Morralla PhD student Julia Pose Utrilla Master s student Noelia Sofía de León Reyes Master s student Aurelio del Pino Beitia Master s student Raquel López Labiano Student Esther Otero Vidal Technician 160

167 2014 Annual Report Teresa Iglesias Vacas Group Summary For the last years, our research group has been focussed on the study of PKD (Protein Kinase D) and its substrate Kidins220 (Kinase D interacting substrate of 220 kda, also known as ankyrin repeat-rich membrane spanning or ARMS), in order to define their role in neuronal physiopathology and their participation in the molecular mechanisms involved in neurodegenerative processes. In a recent publication we found an increase in Kidins220 levels in brain necropsies from Alzheimer s disease (AD) patients, where it accumulates with tau. We identified some of the molecular mechanisms involved, where the kinase GSK-3β is a key regulatory component. During 2014, we have continued studying the role of GSK-3β in the alterations of the cortical circuits that occur in AD, as part of the development of an internal collaborative project. In this same context, we have started the study of identification and characterization of several molecular mechanisms that modulate Kidins220 stability and the effects produced by variations of Kidins220 protein levels on neuronal survival and death, as well as on the appearance of axonal and synaptic alterations. In addition, we have designed a neuroprotective strategy based on the use of peptides with possible therapeutic applications for excitotoxicity, a type of neuronal death elicited by acute or chronic exposure of N-methyl-d-aspartate receptors (NMDARs) to high concentrations of glutamate that takes place in AD and other neurodegenerative disorders. Finally, regarding PKD, we have found that this kinase associates with, phosphorylates and activates neuronal and endothelial nitric oxide synthases (nnos, Sánchez-Ruiloba et al., 2014; and enos, Aicart-Ramos et al., 2014). This activation leads to the production of nitric oxide (NO), a molecule known to play a dual role in the nervous system: on one hand it is crucial for physiological processes while an excessive production under pathological conditions contributes greatly to neurodegenerative processes taking place in disorders such as Parkinson s and Alzheimer s disease. Keywords Kidins220/ARMS, Protein Kinase D (PKD), GSK3, Nitric Oxide Synthase (NOS), Neuroprotection, Neurotoxicity, Alzheimer, Parkinson Publications Aicart-Ramos C, Sanchez-Ruiloba L, Gomez-Parrizas M, Zaragoza C, Iglesias T, Rodriguez-Crespo I. Protein kinase D activity controls endothelial nitric oxide synthesis. Journal of cell science. 2014;127(Pt 15): Epub 2014 Jun 13. Sanchez-Ruiloba L, Aicart-Ramos C, Garcia-Guerra L, Pose-Utrilla J, Rodriguez-Crespo I, Iglesias T. Protein kinase D interacts with neuronal nitric oxide synthase and phosphorylates the activatory residue serine PloS one. 2014;9(4):e Epub 2014 Apr 16. Leon G, Sanchez-Ruiloba L, Perez-Rodriguez A, Gragera T, Martinez N, Hernandez S, et al. Shoc2/Sur8 protein regulates neurite outgrowth. PloS one. 2014;9(12):e Epub 2014 Dec 16. Garcia-Guerra L, Vila-Bedmar R, Carrasco-Rando M, Cruces-Sande M, Martin M, Ruiz-Gomez A, et al. Skeletal muscle myogenesis is regulated by G protein-coupled receptor kinase 2. Journal of molecular cell biology. 2014;6(4): Epub 2014 Jun

168 Research Projects Code: COST Action BM1001: ECMNET Title: Brain Extracellular Matrix in Health and Disease Principal Investigator: Alexander Dityatev CIBERNED s collaboration: Yes Groups CIBERNED: G408; G111 Type: International Funding Agency: COST (European Coordination in Science and Technology) European Office Funding: N.D. Duration: Code: SAF Title: Participation of PKD and its substrate, Kidins220, in the molecular pathways of neuronal survival and neurodegeneration Principal Investigator: Teresa Iglesias Vacas Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: Code: S2010_BMD Title: Redes de señalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas Principal Investigator: Jose González Castaño CIBERNED s collaboration: Yes Groups CIBERNED: G401; G104; G111; G307; G409 Other CIBER s collaboration: Yes (CIBERER) Type: International Funding Agency: Comunidad de Madrid Funding: Duration: Code: 2013/07A Title: Papel de GSK-3 β en las alteraciones de los circuitos corticales que ocurren en la enfermedad de Alzheimer Principal Investigator: Teresa Iglesias Vacas CIBERNED s collaboration: Yes Groups CIBERNED: G401; G403; G111; G504; G307 Type: International Funding Agency: CIBERNED Funding: Duration: Program 2 162

169 Group Jaime Kulisevsky Bojarsky Program 2 CONTACT DETAILS Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Director de Bojarsky la Unidad Group de Trastornos Jaime Kulisevsky de Movimiento Bojarsky Group Jaim Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Servicio Bojarsky de Neurología Group Jaime Kulisevsk Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Hospital de Group la Santa Jaime Creu Kulisevsky i Sant PauBojarsky Group Sant Antoni Maria Claret, 167 Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsk Barcelona (Spain) Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Phone: +34 Jaime 935 Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Fax: Bojarsky Group 872 Jaime Kulisevsk jkulisevsky@santpau.cat Bo Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime Kulisevsky Bojarsky Group Jaime LIST OF PERSONNEL Jaime Kulisevsky Bojarsky Principal Investigator Antonia Campolongo Perillo Researcher Carmen García Sánchez Researcher Alexandre Gironell Carreró Researcher Javier Pagonabarraga Mora Researcher Berta Pascual Sedano Researcher Carles Roig Arnall Researcher Fabián Arenas Ríos Researcher María Victoria Sosti Sosa Researcher Ramón Fernández de Bobadilla PhD student Juan Marín Lahoz PhD student Saúl Martínez Horta PhD student Jesús Pérez Pérez PhD student 163

170 Summary Directed by Jaime Kulisevsky MD, PhD (Movement Disorders Unit, Sant Pau Hospital, Biomedical Research Institute), we focus on translational research of Parkinson s Disease (PD) and other Movement Disorders. During 2014 we initiated new projects and different objectives have been achieved: Genetic risk factors for cognitive impairment. Ongoing analysis of mutations in the glucocerebrosidase gene (GBA) after confirming 6 times increase of the risk of dementia in PD. Apathy in PD. We suggested a higher degree of degeneration of the mesocorticolimbic pathway in apathetic PD patients after showing impairment of a neurophysiological marker of sensitivity to reward (Journal Neuroscience, 2014). Cognition in PD -- Ongoing longitudinal study of patients <5 years of evolution to refine motor, cognitive, emotional and behavioral predictors of cognitive impairment. -- Validation of cognitive (PD-CRS) and functional impairment scales (PD-CFRS) with the new diagnostic criteria for mild cognitive impairment (MCI) in PD (paper in preparation). -- Development of an alternative version of the PD-CRS for reassessment in clinical trials (manuscript in preparation). Predicting the emergence of impulse control disorders (ICD) in PD. Ongoing prospective study to identify cognitive, clinical and neurophysiological characteristics as potential markers of risk for development of ICDs. A 10-years prospective longitudinal study of cognition and quality of life in PD patients underwent bilateral subthalamic deep brain stimulation. Presently in process of analysis of the database. Music-therapy and Biofeedback -- Double-blind study, effects of biofeedback training by Think Interfaces on memory and attention in ADHD patients. -- Double-blind study of effects of music-therapy and cognitive stimulation on abnormal gait in PD. Huntington s disease (HD) -- Consolidation of a Clinical Multidisciplinary Unit on HD. -- Group s IP designed Spanish coordinator of the International Study Enroll-HD: A Prospective Study in a Global Registry Huntington s Disease Cohort (ClinicalTrials.gov Identifier: NCT ). -- Leadership of the project of development of cognitive assessment tools Cognitive Phenotype Working Group of the European Huntington s Disease Network (EHDN). -- Participation in the Task-force for designing a new functional assessment Furst We initiated two lines of research: a) characterization of a BACHD rat model (see section on translational research); b) neuroradiological, neurophysiological and cognitive/behavioural biomarkers in pre-clinical HD stage. Tremor -- Development of a Smartphone online system for neurophysiological studies of tremor. -- Retrospective staging of severity of essential tremor through the glass scale. Dystonia. Participation on the Spanish Multicenter Project Data Collection of the use of Botulinum Toxin and other drugs in Dystonia. MRI study of Tremor-ataxia syndrome associated with fragile X premutation (FXTAS). We observed Program 2 164

171 2014 Annual Report Jaime Kulisevsky Bojarsky Group that the onset of symptoms is associated with loss of cortical thickness in medial temporal lobe and dorsolateral prefrontal and dorsomedial cortex (manuscript in preparation). Translational Research - Experimental Laboratory -- Transgenic rat model of Huntington (BACHD): 12-month longitudinal study of motor, emotional and cognitive disorders. -- Rat model: study of the impact of the degeneration of the noradrenergic system -additional to the dopaminergic lesion- in motor and cognitive response associated with antiparkinsonian drugs. International Collaborations The IP is a member of: -- International Movement Disorders Society (MDS) Task Force for Developing Rating Scales in PD (Sub-committee for Cognitive Evaluation) and member of the Task Force for Developing the MDS-UPDRS (since 2005). -- MDS International Task Force for Diagnosis of Mild Cognitive Impairment in PD (since 2010). -- International Study Group for the validation of MDS PD-MCI criteria ( Mild Cognitive Impairment Validation of PD-MCI Study Group ). Keywords Parkinson, Cognition, Mild Cognitive Impairment, Huntington, Distonia, FxTAS, Essential Tremor, Animal models of parkinsonism Publications Martinez-Horta S, Riba J, de Bobadilla RF, Pagonabarraga J, Pascual-Sedano B, Antonijoan RM, et al. Apathy in Parkinson's disease: neurophysiological evidence of impaired incentive processing. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(17): de Quintana-Schmidt C, Pascual-Sedano B, Alvarez-Holzapfel MJ, Gironell A, Leidinger A, Benito N, et al. Complications related with implanted devices in patients with Parkinson's disease treated with deep brain stimulation. A study of a series of 124 patients over a period of 16 years. Revista de neurologia. 2014;59(2): Pagonabarraga J, Kulisevsky J. Dopaminergic treatment in Parkinson's disease: what has each therapeutic family got to offer? Revista de neurologia. 2014;58(1): Gasso P, Mas S, Papagianni K, Ferrando E, de Bobadilla RF, Arnaiz JA, et al. Effect of CYP2D6 on risperidone pharmacokinetics and extrapyramidal symptoms in healthy volunteers: results from a pharmacogenetic clinical trial. Pharmacogenomics. 2014;15(1): Campolongo A, Marquez Rebollo MC, Alcaraz Escribano ML. Enfermedad de Parkinson y parkinsonismo: cuidados de enfermería Perez Perez J, Kulisevsky J. Enfermedad de Parkinson y transtornos del sueño Campolongo A, Fernandez de Bobadilla R, Pascual-Sedano B, Kulisevsky J. Guía de práctica clínica para enfermería digestiva en pacientes con enfermedad de Parkinson portadores de bomba de levodopa en infusión intraduodenal. EGEH. Enfermeria en Gastroenterología y Hepatología. 2014; 14. Emre M, Poewe W, De Deyn PP, Barone P, Kulisevsky J, Pourcher E, et al. Long-term safety of rivastigmine in parkinson disease dementia: an open-label, randomized study. Clinical neuropharmacology. 2014;37(1):

172 Martinez-Horta S, Kulisevsky J. Mild Cognitive Impairment in Parkinson's Disease. In: Reichmann H, Ed. Neuropsychiatric symptoms of movement disorders. Springer; p Gironell A. The GABA Hypothesis in Essential Tremor: Lights and Shadows. Tremor and other hyperkinetic movements (New York, N.Y.). 2014;4:254. Epub 2014 Jul 16. Cattaneo G, Calabria M, Marne P, Gironell A, Abutalebi J, Costa A. The role of executive control in bilingual language production: A study with Parkinson's disease individuals. Neuropsychologia. 2015;66: Epub 2014 Nov 11. Rami L, Mollica MA, Garcia-Sanchez C, Saldana J, Sanchez B, Sala I, et al. The Subjective Cognitive Decline Questionnaire (SCD-Q): a validation study. Journal of Alzheimer's disease: JAD. 2014;41(2): Marras C, Tröster AI, Kulisevsky J, Stebbins GT. The tools of the trade: a state of the art "How to Assess Cognition" in the patient with Parkinson's disease. Movement disorders: official journal of the Movement Disorder Society. 2014;29(5): Eberling J, Vincent L, Goldman JG, Weintraub D, Kulisevsky J, Marras C, et al. Therapeutic development paths for cognitive impairment in Parkinson's disease: report of a regulatory roundtable. Journal of Parkinson's disease. 2014;4(4): Gironell A, Martinez-Horta S, Aguilar S, Torres V, Pagonabarraga J, Pascual-Sedano B, et al. Transcranial direct current stimulation of the cerebellum in essential tremor: a controlled study. Brain stimulation. 2014;7(3): Epub 2014 Feb 6. Tilley BC, LaPelle NR, Goetz CG, Stebbins GT; MDS-UPDRS Task Force. Using cognitive pretesting in scale development for Parkinson's disease: the Movement Disorder Society Unified Parkinson's Disease Rating Scale (MDS-UPDRS) example. Journal of Parkinson's disease. 2014;4(3): Matias-Guiu JA, Fernandez de Bobadilla R, Escudero G, Perez-Perez J, Cortes A, Morenas-Rodriguez E, et al. Validation of the Spanish version of Addenbrooke's Cognitive Examination III for diagnosing dementia. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Jul 4. Matias-Guiu JA, Fernandez-Bobadilla R. Validation of the Spanish-language version of Mini- Addenbrooke's Cognitive Examination as a dementia screening tool. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Dec 16. Spataro N, Calafell F, Cervera-Carles L, Casals F, Pagonabarraga J, Pascual-Sedano B, et al. Mendelian genes for Parkinson's disease contribute to the sporadic forms of the disease. Human molecular genetics. [Epub ahead of print]. Epub 2014 Dec 11. Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agundez JA, Jimenez-Jimenez FJ, Ross OA, et al. TREM2 R47H variant and risk of essential tremor: A cross-sectional international multicenter study. Parkinsonism & related disorders. [Epub ahead of print]. Epub 2014 Dec 24. Geurtsen GJ, Hoogland J, Goldman JG, Schmand BA, Tröster AI, Burn DJ, et al. Parkinson's disease mild cognitive impairment: application and validation of the criteria. Journal of Parkinson's disease. 2014;4(2): Tolosa E, Hernandez B, Linazasoro G, Lopez-Lozano JJ, Mir P, Marey J, et al. Efficacy of levodopa/ carbidopa/entacapone versus levodopa/carbidopa in patients with early Parkinson's disease experiencing mild wearing-off: a randomised, double-blind trial. Journal of neural transmission. 2014;121(4): Pagonabarraga J, Soriano-Mas C, Llebaria G, Lopez-Solà M, Pujol J, Kulisevsky J. Neural correlates of minor hallucinations in non-demented patients with Parkinson's disease. Parkinsonism & related disorders. 2014;20(3): Program 2 166

173 2014 Annual Report Jaime Kulisevsky Bojarsky Group Research Projects Code: 2011 FI-DGR Title: Contracte AGAUR2010 FI-DGR 2010 Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: Others Funding Agency: AGAUR Funding: Duration: Code: JCI Title: Contrato Juan de la Cierva 2011 Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: International Funding Agency: Ministerio de Economia y Competividad Funding: Duration: 2014 Code: MIA Title: Contrato MIA 2013 Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: Others Funding Agency: Fundación Privada Hospital de la Santa Creu i Sant Pau Funding: Duration: Code: PI12/03005 Title: Desarrollo, validación y estudio de la sensibilidad al cambio de una version alternativa de la Parkinson's Disease - Cognitive Rating Scale Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: Code: Title: Early implementation of music thrapy in rehabilitation of Aphasic patients after acute adquired brain injury /Efectivity of music therapy in the rehabilitation of acute aphasic patients Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: Private Funding Agency: Fundació La Marató de TV3 Funding: Duration: Code: Telemarató 2011 Lesiones medulares y cerebrales Title: Efectividad de la musicoterapia en la rehabilitación en pacientes afásicos agudos Principal Investigator: Carmen García Sánchez Groups CIBERNED: Type: National Funding Agency: Fundació La Marató de TV3 Funding: Duration: Code: AC638 Title: HENUFOOD Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: National Funding Agency: CENIT Funding: Duration: Code: 2014 SGR 1203 Title: Group de Trastornos del Movimiento Hospital de la Santa Creu i Sant Pau Principal Investigator: Jaime Kulisevsky Groups CIBERNED: Type: International Funding Agency: Agència de Gestió d'ajuts Universitaris i de Recerca (AGAUR) Funding: Duration:

174 PhD Dissertations Author: Carme Serrano Munuera Title: Descripción de una nueva ataxia espinocerebelosa dominante con movimientos oculares verticales alterados ligada a un nuevo locus Date: 3rd November 2014 Supervisor: Jaime Kulisevsky Bojarski Program 2 168

175 Group José Luis Labandeira García Program 2 CONTACT DETAILS Group José Luis Labandeira García Group Center for José Research Luis Labandeira in Molecular García Medicine Group and José Chronic Luis Labandeira Diseases (CIMUS) García Group José Lu Labandeira García Group José Luis Labandeira García Group José Universidad Luis Labandeira Santiago García de Group Compostela José Luis Labandeir Avenida Barcelona, 22 García Group José Luis Labandeira García Group José Luis Labandeira García Group José Luis Labandeira García Grou Santiago de Compostela (Spain) José Luis Labandeira García Group José Luis Labandeira García Phone: Group +34 José 881 Luis 812 Labandeira 223 / García 471Group José Lu Labandeira García Group José Luis Labandeira García Group José Luis Labandeira Fax: García Group José Luis Labandeir joseluis.labandeira@usc.es García Group José Luis Labandeira García Group José Luis Labandeira García Group José Luis Labandeira García Grou LIST OF PERSONNEL José L. Labandeira García Principal Investigator, Full Professor Maria J. Guerra Seijas Full Professor Ramón Soto Otero Full Professor Estefanía Méndez Álvarez Full Professor Jannette Rodríguez Pallares Associate Professor Ana M. Muñoz Patiño Associate Professor Ana I. Rodríguez Pérez Associate Professor Carmen Díaz Ruiz Professor Begoña Villar Cheda Associate Professor Juan A. Parga Martin Postdoctoral researcher Pablo Garrido Gil Postdoctoral researcher Ana Borrajo López Predoctoral researcher A. Domínguez Meijide Predoctoral researcher Rita Valenzuela Limiñana Predoctoral researcher Iria Novoa Pérez Specialist Technician Pilar Aldrey García Specialist Technician J.A. Trillo Franco Specialist Technician 169

176 Summary We have studied the role of the basal ganglia renin-angiotensin system (RAS) in dopamine neuron vulnerability and progression of the dopaminergic degeneration, and the possible manipulation of RAS activity as neuroprotective strategy. Over the last few years, a considerable number of studies have shown that local /tissue RAS play a major role in aging and aging-related degenerative processes in several tissues including brain tissue. Important functional interactions between RAS and dopamine and dopamine receptors have been observed in several peripheral tissues, particularly in the cardiovascular and renal systems, in which alteration of this interaction plays a major role in the development of degenerative processes. We have shown an important functional interaction between the local RAS and the nigrostriatal system, as well as a major role of the nigral RAS in progression of dopaminergic degeneration. Over the last year, we have studied the RAS in the nigra and its possible role in neurodegeneration and neuroinflammation, and the intracellular pathways involved in the effects of the local RAS on the dopaminergic vulnerability. In the nigra, we have shown interactions between RAS and Rho-Kinasa activity, and their possible interest for neuroprotective strategies. We have investigated L-DOPA-induced dyskinesias in animal models of parkinsonism, as well as potential treatments. In collaboration with other groups of Ciberned, we have studied the neuroprotective strategies for cell therapy in animal models of parkinsonism. Keywords Neurodegeneration, Neuroprotection, Neuroinflammation, Parkinson, Aging, Angiotensin, Cell therapy Publications Sierra S, Luquin N, Rico AJ, Gomez-Bautista V, Roda E, Dopeso-Reyes IG, et al. Detection of cannabinoid receptors CB1 and CB2 within basal ganglia output neurons in macaques: changes following experimental parkinsonism. Brain structure & function. [Epub ahead of print]. Epub 2014 Jun 28. Farre D, Munoz A, Moreno E, Reyes-Resina I, Canet-Pons J, Dopeso-Reyes IG, et al. Stronger Dopamine D1 Receptor-Mediated Neurotransmission in Dyskinesia. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Oct 26. Villar-Cheda B, Dominguez-Meijide A, Valenzuela R, Granado N, Moratalla R, Labandeira-Garcia JL. Aging-related dysregulation of dopamine and angiotensin receptor interaction. Neurobiology of aging. 2014;35(7): Epub 2014 Jan 17. Munoz A, Garrido-Gil P, Dominguez-Meijide A, Labandeira-Garcia JL. Angiotensin type 1 receptor blockage reduces l-dopa-induced dyskinesia in the 6-OHDA model of Parkinson's disease. Involvement of vascular endothelial growth factor and interleukin-1β. Experimental neurology. 2014;261: Epub 2014 Aug 23. Labandeira-Garcia JL, Garrido-Gil P, Rodriguez-Pallares J, Valenzuela R, Borrajo A, Rodriguez-Perez AI. Brain renin-angiotensin system and dopaminergic cell vulnerability. Front Neuroanat. 2014;8:67. Epub 2014 Jul 8. Rodriguez-Perez AI, Borrajo A, Valenzuela R, Lanciego JL, Labandeira-Garcia JL. Critical period for dopaminergic neuroprotection by hormonal replacement in menopausal rats. Neurobiology of aging. 2015;36(2): Epub 2014 Nov 1. Program 2 170

177 2014 Annual Report José Luis Labandeira García Group Borrajo A, Rodriguez-Perez AI, Villar-Cheda B, Guerra MJ, Labandeira-Garcia JL. Inhibition of the microglial response is essential for the neuroprotective effects of Rho-kinase inhibitors on MPTPinduced dopaminergic cell death. Neuropharmacology. 2014;85:1-8. Epub 2014 May 27. Rodriguez-Perez AI, Borrajo A, Rodriguez-Pallares J, Guerra MJ, Labandeira-Garcia JL. Interaction between NADPH-oxidase and Rho-kinase in angiotensin II-induced microglial activation. Glia. 2015;63(3): Epub 2014 Nov 6. Labandeira-Garcia JL, Rodriguez-Perez AI, Villar-Cheda B, Borrajo A, Dominguez-Meijide A, Guerra MJ. Rho Kinase and Dopaminergic Degeneration: A Promising Therapeutic Target for Parkinson's Disease. The Neuroscientist: a review journal bringing neurobiology, neurology and psychiatry. [Epub ahead of print]. Epub 2014 Oct 16. Borrajo A, Rodriguez-Perez AI, Diaz-Ruiz C, Guerra MJ, Labandeira-Garcia JL. Microglial TNF-α mediates enhancement of dopaminergic degeneration by brain angiotensin. Glia. 2014;62(1): Dominguez-Meijide A, Villar-Cheda B, Garrido-Gil P, Sierrra-Paredes G, Guerra MJ, Labandeira-Garcia JL. Effect of chronic treatment with angiotensin type 1 receptor antagonists on striatal dopamine levels in normal rats and in a rat model of Parkinson's disease treated with L-DOPA. Neuropharmacology. 2014;76(A): Research Projects Code: RD12/0019/0020 Title: Combined protective/restorative cell-mediated strategies for neurodegenerative diseases Red de Terapia celular. NeuroTERCEL Principal Investigator: José Luis Labandeira García CIBERNED s collaboration: Yes Groups CIBERNED: G301; G102; G113; G208; G105; G607 Type: National Funding Agency: ISCIII Funding: Duration: Code: GRC2014/002 Title: Consolidación y estructuración de Groups de referencia competitiva Principal Investigator: José Luis Labandeira García Groups CIBERNED: Type: International Funding Agency: Conselleria de Educación. Xunta de Galicia Funding: Duration: Code: R2014/50 Title: Red Gallega de Terapia Celular Principal Investigator: Clara Alvarez (IP Nodo: J.L. Labandeira) Groups CIBERNED: Other CIBER s collaboration: Yes (CIBERBBN, CIBEROBN) Tipo: CCAA Funding Agency: Conselleria de Educación. Xunta Galicia Funding: Duration:

178 PhD Dissertations Author: Rita Valenzuela Limiñana Title: Sistema Renina-Angiotensina tisular/extracelular y sistema Renina-Angiotensina intracrino/intracelular en la Sustancia Negra. Posible papel en la enfermedad de Parkinson Date: 5th December 2014 Supervisor: José Luis Labandeira García Program 2 172

179 Group José Luis Lanciego Pérez Program 2 CONTACT DETAILS Group José Luis Lanciego Pérez Group José Luis Lanciego Pérez Group José Área Luis de Lanciego Neurociencias, Pérez Group CIMA José Luis Lancieg Pérez Group José Luis Lanciego Pérez Group José Luis Lanciego Pérez Group José Luis Avenida Lanciego Pio XII, Pérez 55Group José Lu Lanciego Pérez Group José Luis Lanciego Pérez Group José Luis Lanciego Pérez Pamplona, Group José Navarra Luis (Spain) Lanciego Pérez Group jlanciego@unav.es José Luis Lanciego Pérez Group José Luis Lanciego Pérez Group José Luis Lanciego Pérez Group José Luis Lanciego Pére LIST OF PERSONNEL José Luis Lanciego Pérez Principal Investigator Alberto José Rico Martín PhD Iria González Dopeso-Reyes PhD Gloria González Aseguinolaza PhD Eva Martínez Pinilla PhD José Diego Pignataro López PhD student Elvira Roda Recalde Lab Technician 173

180 Summary Our research group is mainly engaged in the study of the neurobiology of Parkinson disease, with a multiple focus on a number of different topics related to the pathophysiology of this disease, as follows: Analysis of brain circuits underlying the pathophysiology of Parkinson disease. By using relevant animal models of the disease, we study the ongoing changes in basal ganglia circuits following dopaminergic neurodegeneration. Study of G-protein-coupled receptors (GPCRs) in Parkinson disease. Our main interest is to address the presence of different GPCRs in basal ganglia nuclei. Among others, we focus on endocannabinoid receptors 1 and 2 (CB1 and CB2), orphan receptor GPR55, adenosine type 2A receptors (A2A) and dopamine receptors types 1 and 2 (D1 and D2). All studies are carried out in control and parkinsonian animals. More specifically, we are very much interested in elucidating which GPCRs are making heteromers, with the ultimate goal of addressing whether these GPCR heteromers may be considered as feasible pharmacological targets. Cell and gene therapies in Parkinson disease. Together with a number of European collaborators, we have recently started a research project focused on the use of direct reprogramming techniques converting skin fibroblast onto dopaminergic neurons that will further be used for autologous transplantation in relevant animal models of parkinsonism. Moreover, a number of gene therapybased approaches are currently under implementation to further manipulate basal ganglia circuits of interest. Keywords Basal ganglia, Parkinson s disease, GPCR receptors, Gene therapy, Non-human primates Publications Zamarbide M, Etayo-Labiano I, Ricobaraza A, Martinez-Pinilla E, Aymerich MS, Lanciego JL, et al. GPR40 activation leads to CREB and ERK phosphorylation in primary cultures of neurons from the mouse CNS and in human neuroblastoma cells. Hippocampus. 2014;24(7): Epub 2014 Feb 27. Dopeso-Reyes IG, Rico AJ, Roda E, Sierra S, Pignataro D, Lanz M, et al. Calbindin content and differential vulnerability of midbrain efferent dopaminergic neurons in macaques. Frontiers in neuroanatomy. 2014;8:146. Epub 2014 Dec 3. Martinez-Pinilla E, Reyes-Resina I, Onatibia-Astibia A, Zamarbide M, Ricobaraza A, Navarro G, et al. CB1 and GPR55 receptors are co-expressed and form heteromers in rat and monkey striatum. Experimental neurology. 2014;261: Epub 2014 Jun 23. Sierra S, Luquin N, Rico AJ, Gomez-Bautista V, Roda E, Dopeso-Reyes IG, et al. Detection of cannabinoid receptors CB1 and CB2 within basal ganglia output neurons in macaques: changes following experimental parkinsonism. Brain structure & function. [Epub ahead of print]. Epub 2014 Jun 28. Brugarolas M, Navarro G, Martinez-Pinilla E, Angelats E, Casado V, Lanciego JL, et al. G-proteincoupled receptor heteromers as key players in the molecular architecture of the central nervous system. CNS neuroscience & therapeutics. 2014;20(8): Epub 2014 May 9. Cerri S, Levandis G, Ambrosi G, Montepeloso E, Antoninetti GF, Franco R, et al. Neuroprotective potential of adenosine A2A and cannabinoid CB1 receptor antagonists in an animal model of Program 2 174

181 2014 Annual Report José Luis Lanciego Pérez Group Parkinson disease. Journal of neuropathology and experimental neurology. 2014;73(5): Garcia-Barroso C, Ugarte A, Martinez M, Rico AJ, Lanciego JL, Franco R, et al. Phosphodiesterase inhibition in cognitive decline. Journal of Alzheimer's disease: JAD. 2014;42(Suppl 4):S Afonso-Oramas D, Cruz-Muros I, Castro-Hernandez J, Salas-Hernandez J, Barroso-Chinea P, Garcia- Hernandez S, et al. Striatal vessels receive phosphorylated tyrosine hydroxylase-rich innervation from midbrain dopaminergic neurons. Frontiers in neuroanatomy. 2014;8:84. Epub 2014 Aug 26. Farre D, Munoz A, Moreno E, Reyes-Resina I, Canet-Pons J, Dopeso-Reyes IG, et al. Stronger Dopamine D1 Receptor-Mediated Neurotransmission in Dyskinesia. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Oct 26. Rodriguez-Perez AI, Borrajo A, Valenzuela R, Lanciego JL, Labandeira-Garcia JL. Critical period for dopaminergic neuroprotection by hormonal replacement in menopausal rats. Neurobiology of aging. 2015;36(2): Epub 2014 Nov 1. Bonaventura J, Rico AJ, Moreno E, Sierra S, Sanchez M, Luquin N, et al. L-DOPA-treatment in primates disrupts the expression of A(2A) adenosine-cb(1) cannabinoid-d(2) dopamine receptor heteromers in the caudate nucleus. Neuropharmacology. 2014;79: Pinna A, Bonaventura J, Farre D, Sanchez M, Simola N, Mallol J, et al. L-DOPA disrupts adenosine A(2A)- cannabinoid CB(1)-dopamine D(2) receptor heteromer cross-talk in the striatum of hemiparkinsonian rats: biochemical and behavioral studies. Experimental neurology. 2014;253: Research Projects Code: BFU Title: Coexpresión de receptores de adenosina 2A y de endocannabinoides 1 y 2 en los núcleos de salida de los ganglios basales en el modelo de enfermedad de Parkinson en monos Principal Investigator: José Luis Lanciego Groups CIBERNED: Type: International Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: NARSAD Title: Immunotoxin-mediated selective removal of specific basal ganglia pathways in the monkey model of Parkinson's disease Principal Investigator: José Luis Lanciego Groups CIBERNED: Type: International Funding Agency: Brain & Behavior Research Foundation Funding: Duration: Code: COEN-PATHFINDER Title: In vivo neuronal cell reprogramming for a new regenerative approach in Parkinson's disease Principal Investigator: José Luis Lanciego (CIBERNED, Spain) Groups CIBERNED: Type: International Funding Agency: ISCIII-CIBERNED Funding: Duration: Code: ERC Advanced Title: New Experimental therapeutic approaches for Parkinson's disease by direct DA neuronal reprogramming Principal Investigator: José Luis Lanciego (CIBERNED, Spain)(Co-IP) Groups CIBERNED: Type: International Funding Agency: European Research Council (ERC Advanced Grants) Funding: Duration:

182 Group José López Barneo Program 2 CONTACT DETAILS oup José López Barneo Group José López Barneo Group José López Instituto Barneo de Biomedicina Group José de López Sevilla Barneo Group sé López Barneo Group José López Barneo Group José López Laboratorio Barneo Group Neurobiología José López celular Barneo y biofísica Group José López rneo Group José López Barneo Group José López Barneo Campus Group Hospital José Universitario López Barneo Virgen Group del Rocío José López Barneo Avenida Manuel Siurot, s/n oup José López Barneo Group José López Barneo Group José López Barneo Group José López Barneo Group Sevilla (Spain) sé López Barneo Group José López Barneo Group José López Barneo Group Phone: José López Barneo 923 Group 007 José López rneo Group José López Barneo Group José López Barneo Group José López Barneo Fax: +34 Group José 101López Barneo lbarneo@us.es oup José López Barneo Group José López Barneo Group José López Barneo Group José López Barneo Group LIST OF PERSONNEL José López Barneo Principal Investigator, Full Professor Aida Platero Luengo Predoctoral researcher Candela Caballero Erazo Río Hortega Researcher Carmen Calero Acuña Río Hortega Researcher David Macías Gutiérrez Postdoctoral researcher Francisco Javier Villadiego Luque Postdoctoral researcher Gloria Paula García Flores Technician Helia Sarmiento Soto Technician Ignacio Arias Mayenco Technician Ivette López López Technician José Antonio Rodríguez Gómez Researcher Juan José Toledo Aral Researcher Lin Gao Researcher María del Carmen Fernandez- Aguera Rodríguez Predoctoral researcher Nela Suárez Luna Technician Pablo Mir Rivera Physician Patricia González Rodríguez Postdoctoral researcher Patricia Ortega Sáenz Researcher Pilar Gómez Garre Researcher Ricardo Pardal Redondo Researcher Victoria Bonilla Henao Specialist Technician Xavier d Anglemont de Tassigny Postdoctoral researcher 176

183 2014 Annual Report José López Barneo Group Summary In 2014, our group has developed three research lines: Pathogenesis of Parkinson s disease (PD) We are currently studying the early stages of neurodegenerative diseases, such as Parkinson s disease (PD), using three transgenic animal models carrying deletions in distinct mitochondrial complexes. We have developed several animal models that are now in the characterization step to i) confirm the critical role of the endogenous neurotrophic glial derived factor (GDNF) on the maintenance of the nigrostriatal pathway, ii) clarify the nature of the GDNF-producing neurons, iii) study the difference at the molecular level of the intracellular mechanisms leading to the synthesis and secretion of GDNF in the striatum. For this, we are also setting up a cell sorting protocol to compare genomic and proteomic patterns of the striatal parvalbumin neurons (GDNF+) from the same neurons located in the nearby cortex (negative for GDNF). This will help to understand the regulation of endogenous GDNF synthesis. In parallel to the animal models, we are continuing the study on genetic alterations related to PD and movement disorders. Additionally, we are monitoring the putative cortical activity changes in PD patients by using transcranial magnetic stimulation. Cell therapy in PD In collaboration with the company Axontherapix, transplantations of carotid body (CB)-derived neurospheres from rodents have been succesfully performed in Parkinsonian animal models (obtained by chronic administration of MPTP). These CB neurospheres provided a clear neuroprotective action on the nigrostriatal pathway as well as recovery of the striatal dopaminergic terminals. This effect can be explained by a trophic support presumably mediated by GDNF, which is highly produced by these neurospheres. In parallel, human CB transplantations have been performed in PD mice to study the trophic effect exerted by GDNF on the nigrostriatal neurons. Indeed, these human CB grafts provided a trophic action on the nigrostriatal pathway. Moreover, various variables (i.e. age and sex of the human donors) have been evaluated on the trophic action exerted by the transplanted CB tissue. In this regard, age represents a limiting factor that reduces the therapeutic efficacy of carotid tissue, and we are now further studying the molecular mechanisms involved in this aspect. In any case, the neurotrophic actions on the nigrostriatal pathway by the human CB and the human CB-derived neurospheres are under writing for publication. Effect of hypoxia on the neurogenic niches in the central and peripheral nervous system As part of this research project, we are investigating the effect of chronic and intermittent hypoxia on the neurogenic niche. We have developed two animal models of chronic and intermittent hypoxia in rodent animals. These models are now in use in combination with several state-of-the-art techniques to analyze the changes occuring in the neurogenic niche at both the central (subventricular zone SVZ) and peripheral (CB) levels. Keywords Hypoxia, Carotid body, Movement disorders, Parkinson s disease, Neurogenesis, GDNF 177

184 Publications Millan-Ucles A, Diaz-Castro B, Garcia-Flores P, Baez A, Perez-Simon JA, Lopez-Barneo J, et al. A conditional mouse mutant in the tumor suppressor SdhD gene unveils a link between p21(waf1/ Cip1) induction and mitochondrial dysfunction. PLoS One. 2014;9(1):e Epub 2014 Jan 20. Sanchez Gomar I, Diaz Sanchez M, Ucles Sanchez AJ, Casado Chocan JL, Ramirez-Lorca R, Serna A, et al. An immunoassay that distinguishes real neuromyelitis optica signals from a labeling detected in patients receiving natalizumab. BMC Neurology. 2014;14:139. Epub 2014 Jul. Jesus S, Gomez-Garre P, Carrillo F, Caceres-Redondo MT, Huertas-Fernandez I, Bernal-Bernal I, et al. Analysis of c.801-2a>g mutation in the DNAJC6 gene in Parkinson's disease in southern Spain. Parkinsonism Relat Disord. 2014;20(2): Epub 2013 Oct 31. Gomez-Garre P, Huertas-Fernandez I, Caceres-Redondo MT, Alonso-Canovas A, Bernal-Bernal I, Blanco-Ollero A, et al. BDNF Val66Met polymorphism in primary adult-onset dystonia: a case-control study and meta-analysis. Movement Disorders. 2014;29(8): Epub 2014 Jun 12. Benitez-Rivero S, Lama MJ, Huertas-Fernandez I, Alvarez de Toledo P, Caceres-Redondo MT, Martin- Rodriguez JF, et al. Clinical features and neuropsychological profile in vascular parkinsonism. Journal of the Neurological Science. 2014;345(1-2): Epub 2014 Jul 28. Macias D, Fernandez-Agüera MC, Bonilla-Henao V, Lopez-Barneo J. Deletion of the von Hippel- Lindau gene causes sympathoadrenal cell death and impairs chemoreceptor-mediated adaptation to hypoxia. EMBO molecular medicine. 2014;6(12): Epub 2014 Nov 10. Olmeda B, Umstead TM, Silveyra P, Pascual A, Lopez-Barneo J, Phelps DS, et al. Effect of hypoxia on lung gene expression and proteomic profile: insights into the pulmonary surfactant response. Journal of proteomics. 2014;101: Epub 2014 Feb 24. Koch G, Porcacchia P, Ponzo V, Carrillo F, Caceres-Redondo MT, Brusa L, et al. Effects of two weeks of cerebellar theta burst stimulation in cervical dystonia patients. Brain Stimulation. 2014;7(4): Epub 2014 May 9. Serna A, Galan-Cobo A, Rodrigues C, Sanchez-Gomar I, Toledo-Aral JJ, Moura TF, et al. Functional inhibition of aquaporin-3 with a gold-based compound induces blockage of cell proliferation. Journal of Cellular Physiology. 2014;229(11): Epub 2014 Nov. Lohmann K, Schmidt A, Schillert A, Winkler S, Albanese A, Baas F, et al. Genome-wide association study in musician's dystonia: a risk variant at the arylsulfatase G locus? Movement Disorders. 2014;29(7): Epub 2013 Dec 26. Montaner J, Jimenez-Hernandez MD, Lopez-Barneo J. How to unfasten the Spanish Stroke Belt? Andalusia chooses research. International journal of stroke: official journal of the International Stroke Society. 2014;9(7): Epub 2014 Jul 15. Gomez-Garre P, Huertas-Fernandez I, Caceres-Redondo MT, Alonso-Canovas A, Bernal-Bernal I, Blanco-Ollero A, et al. Lack of validation of variants associated with cervical dystonia risk: A GWAS replication study. Movement Disorders. 2014;29(14): Epub 2014 Sep 25. Caceres-Redondo MT, Carrillo F, Lama MJ, Huertas-Fernandez I, Vargas-Gonzalez L, Carballo M, et al. Long-term levodopa/carbidopa intestinal gel in advanced Parkinson's disease. J Neurology. 2014;261(3): Epub 2014 Jan 30. Porras-Gonzalez C, Gonzalez-Rodriguez P, Calderon-Sanchez E, Lopez-Barneo J, Urena J. Lowdose combination of Rho kinase and L-type Ca(2+) channel antagonists for selective inhibition of depolarization-induced sustained arterial contraction. European journal of pharmacology. 2014;732: Epub 2014 Mar 27. Kojovic M, Mir P, Trender-Gerhard I, Schneider SA, Parees I, Edwards MJ, et al. Motivational modulation of bradykinesia in Parkinson's disease off and on dopaminergic medication. J Neurology. 2014;261(6): Epub 2014 Apr 1. Program 2 178

185 2014 Annual Report José López Barneo Group Porcacchia P, Palomar FJ, Caceres-Redondo MT, Huertas-Fernandez I, Martin-Rodriguez JF, Carrillo F, et al. Parieto-motor cortical dysfunction in primary cervical dystonia. Brain Stimulation. 2014;7(5): Epub 2014 Jun 23. Gomez-Garre P, Jesus S, Carrillo F, Caceres-Redondo MT, Huertas-Fernandez I, Bernal-Bernal I, et al. Systematic mutational analysis of FBXO7 in a Parkinson's disease population from southern Spain. Neurobiol Aging. 2014;35(3):727.e5-7. Epub 2013 Oct 8. Gomez-Caravaca MT, Caceres-Redondo MT, Huertas-Fernandez I, Vargas-Gonzalez L, Carrillo F, Carballo M, et al. The use of botulinum toxin in the treatment of sialorrhea in parkinsonian disorders. Neurology Science. 2015;36(2): Epub 2014 Sep 20. Huertas-Fernandez I, Garcia-Gomez FJ, Garcia-Solis D, Benitez-Rivero S, Marin-Oyaga VA, Jesus S, et al. Machine learning models for the differential diagnosis of vascular parkinsonism and Parkinson's disease using (123)IFP-CIT SPECT. European journal of nuclear medicine and molecular imaging. 2015;42(1): Epub 2014 Aug 14. Arbizu J, Luquin MR, Abella J, de la Fuente-Fernandez R, Fernandez-Torron R, Garcia-Solis D, et al. Functional neuroimaging in the diagnosis of patients with Parkinsonism: Update and recommendations for clinical use. Revista espanola de medicina nuclear e imagen molecular. 2014;33(4): Epub 2014 Apr 14. Tolosa E, Hernandez B, Linazasoro G, Lopez-Lozano JJ, Mir P, Marey J, et al. Efficacy of levodopa/ carbidopa/entacapone versus levodopa/carbidopa in patients with early Parkinson's disease experiencing mild wearing-off: a randomised, double-blind trial. Journal of neural transmission. 2014;121(4): Platero-Luengo A, Gonzalez-Granero S, Duran R, Diaz-Castro B, Piruat JI, Garcia-Verdugo JM, et al. An O2-sensitive glomus cell-stem cell synapse induces carotid body growth in chronic hypoxia. Cell. 2014;156(1-2): Research Projects Code: SAF2012/39343 Title: Sensibilidad al oxígeno y neurodegeneración Principal Investigator: José López Barneo Groups CIBERNED: Type: National Funding Agency: Ministerio de Economía y Competividad Funding: Duration: Code: SAF Title: Caracterización de los mecanismos intrínsecos implicados en el mantenimiento de las neuronas de la SNPC. Papel de GDNF y de genes modulados por el envejecimiento Principal Investigator: Alberto Pascual Bravo Groups CIBERNED: Type: International Funding Agency: Ministerio de Ciencia e Innovación - FEDER Funding: N.D. Duration: Code: Title: EMTICS (European Multicentre Tics in Children Studies) Principal Investigator: Pablo Mir Rivera Groups CIBERNED: Type: European Funding Agency: Séptimo Program Marco. Comisión Europea. FP7-HEALTH-2011 Funding: Duration: Code: Fundación Mutua Madrileña 13 Title: Estudio de asociación de los genes relacionados con plasticidad neuronal y distonía primaria Principal Investigator: Pablo Mir Rivera Groups CIBERNED: Type: National 179

186 Funding Agency: Fundación Mutua Madrileña Funding: Duration: Code: PAI 2010 (CTS 516) Title: Fisiología Celular y Biofísica Principal Investigator: José López Barneo Groups CIBERNED: Tipo: CCAA Funding Agency: Consejería de Innovación, Ciencia y Empresa. Junta Andalucía Funding: Duration: Code: SAF P Title: Fisiopatología de células madre derivadas de la cresta neural Principal Investigator: Ricardo Pardal Groups CIBERNED: Type: National Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: PIE 2009 Excelencia (P09-CTS-4589) Title: Mecanismos moleculares de formación de tumores en modelos genéticos animales con defectos mitocondriales Principal Investigator: José López Barneo Groups CIBERNED: Tipo: CCAA Funding Agency: Consejería de Innovación, Ciencia y Empresa. Junta Andalucía Funding: Duration: Code: ERC (FP ) Title: Physiology of the adult carotid body stem cell niche Principal Investigator: Ricardo Pardal Groups CIBERNED: Type: European Funding Agency: Comisión de la Comunidad Europea Funding: Duration: Code: FIS 13 (PI13/01461) Title: Plasticidad neuronal en las discinesias inducidas por levodopa en la enfermedad de Parkinson Principal Investigator: Pablo Mir Rivera Groups CIBERNED: Type: National Funding Agency: Instituto de Salud Carlos III. Ministerio Sanidad y Consumo Funding: Duration: Code: Instituto de Salud Carlos III Title: Search and selection of Parkinson s disease candidate and validation by human genetic analysis Principal Investigator: Luis María Escudero Cuadrado Groups CIBERNED: Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: Fundación Marcelino Botín (Nuevo convenio) Title: Sensibilidad al Oxígeno y Neurodegeneración Principal Investigator: José López Barneo Groups CIBERNED: Type: International Funding Agency: Fundación Marcelino Botín Funding: Duration: Code: RD12/0019/0033 Title: Red de Terapia Celular Principal Investigator: Juan José Toledo Aral CIBERNED s collaboration: Yes Groups CIBERNED: G301; G113; G105; G208; G102; G607 Type: Intramural Funding Agency: Red Terapia Celular Funding: Duration: Code: FIS 12 (PI12/02574) Title: Uso de cuerpo carotídeo en terapia celular en la enfermedad de Parkinson Principal Investigator: Juan José Toledo Aral Groups CIBERNED: Program 2 180

187 2014 Annual Report José López Barneo Group Type: National Funding Agency: Instituto de Salud Carlos III. Ministerio Sanidad y Consumo Funding: Duration:

188 Group José Javier Lucas Lozano Program 2 CONTACT DETAILS oup José Javier Lucas Lozano Group José Javier Centro Lucas de Lozano Biología Group Molecular José Javier Severo Lucas Ochoa Lozano CSIC/UAM Group José Javier Lucas zano Group José Javier Lucas Lozano Group José Javier Lucas Lozano Group José Nicolás Javier Lucas Cabrera, Lozano 1 Group José vier Lucas Lozano Group José Javier Lucas Lozano Group José Javier Lucas Lozano Group Madrid José (Spain) Javier Lucas Lozano Phone: / (Lab) oup José Javier Lucas Lozano Group José Javier Lucas Lozano Group José Javier Lucas Lozano Group José Javier Fax: cas Lozano Group José Javier Lucas Lozano Group José Javier Lucas Lozano Group jlucas@cbm.uam.es José Javier Lucas Lozano Group sé Javier Lucas Lozano Group José Javier Lucas Lozano Group José Javier Lucas Lozano Group José Javier Lucas LIST OF PERSONNEL José Javier Lucas Lozano Principal Investigator Raquel Gómez Sintes Postdoctoral Fellow CIBERNED Jorge Rubén Cabrera Heredia Postdoctoral Fellow CIBERNED Marta Fernández Nogales PhD student Alberto Parras Rodríguez PhD student Ivó Hernández Hernández PhD student Elena Llorente Martín Undergrad student Miriam Lucas Santamaría Technician CIBERNED María Santos Galindo Technician 182

189 2014 Annual Report Group José Javier Lucas Lozano Summary Huntington s disease is an autosomal dominant neurodegenerative disease characterized by involuntary movements, psychiatric symptoms and dementia. It is caused by an expansion of the trinucleotide CAG located in exon 1 of the huntingtin gene. In our lab we study the molecular basis of HD through in vitro and in vivo (generating and characterizing transgenic models that could mimic as well as revert the disease) approaches and through studies on human material from patients. Thanks to the human brain postmortem tissue from HD individuals we could demonstrate the imbalance in the tau isoforms with 3 or 4 Microtubule Binging Domains and discover a new histopathological hallmark (the Tau Nuclear Rods or TNRs) in this disease, consisting on the presence of tau in nuclear indentations (Fernandez-Nogales et al., Nat Med. 2014). The utilization of transgenic animals with varying levels of tau allowed us to observe how a moderate decrease in this protein in HD model mice was able to attenuate the disease phenotype. This discovery relates HD with tauopathies, broadening the possible therapies applicable to HD to those generated for the treatment of taupathies, such as Alzheimer s disease. We have also described for the first time the expression of ATF5, an ER stress protein, in adult brain neurons (Brain. 2013;136(Pt 4): ) and its induction with neuroprotective effects in an epilepsy model. The characterization of model of genetic or pharmacological inhibition of GSK-3 has allowed us to observe an HD-like phenotype with motor deficits and neuronal apoptosis (EMBO J. 2007;26(11): ) mediated by Fas/NFAT (J Clin Invest. 2010:120(7): ; PLoS One. 2013;8(8):e70952). Additionally, we have studied the phenotype of a mouse strain with a natural deletion in DISC1, which shows a broad variety of psychiatric disease related behaviors (Gomez-Sintes et al., Front Behav Neurosci. 2014). Keywords Huntington s disease, Transgenic mice, GSK-3, Tau, ER-stress, ATF5, Splicing Publications Formentini L, Pereira MP, Sanchez-Cenizo L, Santacatterina F, Lucas JJ, Navarro C, et al. In vivo inhibition of the mitochondrial H+-ATP synthase in neurons promotes metabolic preconditioning. The EMBO journal. 2014;33(7): Epub 2014 Feb 12. Gomez-Sintes R, Kvajo M, Gogos JA, Lucas JJ. Mice with a naturally occurring DISC1 mutation display a broad spectrum of behaviors associated to psychiatric disorders. Frontiers in behavioral neuroscience. 2014;8:253. Epub 2014 Jul 30. Gomez-Sintes R, Bortolozzi A, Artigas F, Lucas JJ. Reduced striatal dopamine DA D2 receptor function in dominant-negative GSK-3 transgenic mice. European neuropsychopharmacology: the journal of the European College of Neuropsychopharmacology. 2014;24(9): Epub 2014 Jul 21. Ortega Z, Lucas JJ. Ubiquitin-proteasome system involvement in Huntington's disease. Frontiers in molecular neuroscience. 2014;7:77. Epub 2014 Sep 29. Ettcheto M, Junyent F, de Lemos L, Pallas M, Folch J, Beas-Zarate C, et al. Mice Lacking Functional Fas Death Receptors Are Protected from Kainic Acid-Induced Apoptosis in the Hippocampus. Molecular neurobiology. [Epub ahead of print]. Epub 2014 Aug 15. Fernandez-Nogales M, Cabrera JR, Santos-Galindo M, Hoozemans JJ, Ferrer I, Rozemuller AJ, et al. Huntington's disease is a four-repeat tauopathy with tau nuclear rods. Nature medicine. 183

190 2014;20(8): Epub 2014 Jul 20. Casadei N, Pöhler AM, Tomas-Zapico C, Torres-Peraza J, Schwedhelm I, Witz A, et al. Overexpression of synphilin-1 promotes clearance of soluble and misfolded alpha-synuclein without restoring the motor phenotype in aged A30P transgenic mice. Human molecular genetics. 2014;23(3): Research Projects Code: SAF Title: Convergencia de alteraciones génicas, transcripcionales y traduccionales en la enfermedad de Huntington Principal Investigator: José J. Lucas Groups CIBERNED: Type: National Funding Agency: MINECO, Plan Nacional de I+D+I Funding: Duration: Code: HRA_POR/2011/41 Title: The proteasome as a treatment target for preventing seizure induced neuronal death and epilepsy Principal Investigator: David C. Henshall Groups CIBERNED: Type: International Funding Agency: Irish Health Research Board Funding: Duration: Code: NEURO-MIR Title: MicroRNA as novel therapeutic targets and disease biomarkers in Alzheimer's Disease, Frontotemporal dementia and Amyotrophic lateral sclerosis (NEURO-MIR) Principal Investigator: José J. Lucas Groups CIBERNED: Type: International Funding Agency: Iniciativa CoEN Funding: Duration: Program 2 184

191 Group María Ángeles Mena Gómez Program 2 CONTACT DETAILS Group María Ángeles Mena Gómez Group María Ángeles Mena Gómez Group Hospital María Ramón Ángeles y Mena Cajal Gómez Group María Ángeles Mena Gómez Group María Ángeles Mena Gómez Group Carretera MaríaÁngeles de Colmenar Mena km Gómez 9,1 Group Marí Ángeles Mena Gómez Group María Ángeles Mena Gómez Group María Ángeles Mena Madrid Gómez (Spain) Group María Ángele Phone: Mena Gómez Group María Ángeles Mena Gómez Group María Ángeles María Ángeles Mena Gómez Group Marí Fax: Ángeles Mena Gómez Group María Ángeles Mena maria.a.mena@hrc.es Gómez Group María / neurol.hrc@salud.madrid.org María Ángeles Mena Gómez Group Marí Ángeles Mena Gómez Group María Ángeles Mena Gómez Group María o María Ángele LIST OF PERSONNEL María Ángeles Mena Gómez Principal Investigator Justo García de Yébenes Neurologist Guillermo García Ribas Neurologist José Luis López Sendón Neurologist Juan García Caldentey Neurologist Patricia Trigo Neurophychologist María José Casarejos Associate Researcher Juan Perucho Postdoctoral Fellow Conceição Bettencourt Postdoctoral Fellow Raquel Ros Postdoctoral Fellow Carolina Ruiz PhD student Marian Fernández PhD student Andrea Mohedano Nurse Ana Gómez Technician María Paz Muñoz Technician María Ángeles Robles Administrative assistant 185

192 Summary In 2014 our group has maintained the research in neurodegenerative disorders mainly dementias like Alzheimer disease, Parkinson s disease, Progressive supranuclear palsy, Huntington s disease, dystonias, ataxias and paraparexias. Our work centred in: Clinical and molecular characterization of neurodegenerative diseases: Identification of new genes and mutations implicated in these diseases. We continued in collaboration with others groups, Spanish, Portuguese, Dutch and English. We participated in the identification of new genes using the whole-exome sequencing in patients with ataxia and PSP diseases. Clinical studies on neuroprotection compounds in these diseases. This is the case of the clinical trial of SATIVEX drug in Huntington patients. Study in experimental models (transgenic and cell models) of the pathogenic mechanism and the neuroprotective treatments in these diseases. We continued the study of the amyloidogenic role of volatile anesthetics and the role of biomarkers in human cerebrospinal fluid in dementia, the systems of cellular processing of abnormal proteins in models of Parkinson s disease and the role of cannabinoids in Huntington s disease. In the second half of 2014 we have initiated a new line of work, the development of digital multicentric registries for the investigation of neurological diseases and for the evaluation of movement disorders. We have designed a digital registry for Parkinson s disease that is going to be implemented in the next few weeks by the Federation of associations of patients with Parkinson s disease and one method for the quantitative analysis of dyskinesias of the head. Keywords Neurodegeneratives diseases, Neurogenetics, Cell cultures, Transgenics mouses, Neuroprotection Publications Garcia-Ribas G, Lopez-Sendon Moreno JL, Garcia-Caldentey J. Biomarkers in Alzheimer s disease. Revista de neurologia. 2014;58(7): Gonzalo-Gobernado R, Calatrava-Ferreras L, Perucho J, Reimers D, Casarejos MJ, Herranz AS, et al. Liver Growth Factor as a tissue regenerating factor in neurodegenerative diseases. Recent patents on CNS drug discovery. [Epub ahead of print]. Epub 2014 Dec 24. Calatrava-Ferreras L, Gonzalo-Gobernado R, Reimers D, Herranz AS, Jimenez-Escrig A, Diaz-Gil JJ, et al. Neuroprotective role of liver growth factor LGF in an experimental model of cerebellar ataxia. International Journal of Molecular Sciences. 2014;15(10): Epub 2014 Oct 21. Alonso-Canovas A, Lopez-Sendon JL, Buisan J, defelipe-mimbrera A, Guillan M, Garcia-Barragan N, et al. Sonography for diagnosis of Parkinson disease-from theory to practice: a study on 300 participants. Journal of ultrasound in medicine: official journal of the American Institute of Ultrasound in Medicine. 2014;33(12): Casarejos MJ, Perucho J, Lopez-Sendon JL, Garcia de Yebenes J, Bettencourt C, Gomez A, et al. Trehalose improves human fibroblast deficits in a new CHIP-mutation related ataxia. PLoS one. 2014;9(9):e Epub 2014 Sep 26. Program 2 186

193 2014 Annual Report María Ángeles Mena Gómez Group Fernandez-Estevez MA, Casarejos MJ, Lopez Sendon J, Garcia Caldentey J, Ruiz C, Gomez A, et al. Trehalose reverses cell malfunction in fibroblasts from normal and Huntington s disease patients caused by proteosome inhibition. PloS one. 2014;9(2):e Epub 2014 Feb 25. Tolosa E, Litvan I, Höglinger GU, Burn D, Lees A, Andres MV, et al. A phase 2 trial of the GSK-3 inhibitor tideglusib in progressive supranuclear palsy. Movement disorders: official journal of the Movement Disorder Society. 2014; 29(4): Epub 2014 Feb 14. Höglinger GU, Huppertz HJ, Wagenpfeil S, Andres MV, Belloch V, Leon T, et al. Tideglusib reduces progression of brain atrophy in progressive supranuclear palsy in a randomized trial. Movement disorders: official journal of the Movement Disorder Society. 2014;29(4): Epub 2014 Jan 31. Lopez-Sendon Moreno JL, Garcia-Caldentey J, Regidor I, del Alamo M, Garcia de Yebenes J. A 5-year follow-up of deep brain stimulation in Huntington s disease. Parkinsonism & related disorders. 2014;20(2): Bettencourt C, Lopez-Sendon JL, Garcia-Caldentey J, Rizzu P, Bakker IM, Shomroni O, et al. Exome sequencing is a useful diagnostic tool for complicated forms of hereditary spastic paraplegia. Clinical genetics. 2014;85(2): Research Projects Code: S2010/BMD-2308 Title: Neurofarmacología del sistema endocannabinoide: del laboratorio a la clínica Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G303; G304; G305 Other CIBER s collaboration: Yes (CIBERSAM) Tipo: CCAA Funding Agency: Comunidad de Madrid Funding: Duration: Code: CIBERNED 2013/05 Title: Identificación y caracterización molecular de subpoblaciones de receptores cannabinoides en poliglutaminopatías Principal Investigator: Manuel Guzmán Pastor CIBERNED s collaboration: Yes Groups CIBERNED: G201; G303; G304; G305 Type: International Funding Agency: CIBERNED Funding: Duration:

194 Group Rosario Moratalla Villalba Program 2 CONTACT DETAILS oup Rosario Moratalla Villalba Group Rosario Moratalla Villalba Group Rosario Instituto Moratalla Cajal Villalba (CSIC) Group Rosario oratalla Villalba Group Rosario Moratalla Villalba Group Rosario Moratalla Villalba Avenida Group Doctor Rosario Arce, 37 Moratalla Villalba oup Rosario Moratalla Villalba Group Rosario Moratalla Villalba Group Rosario Moratalla Madrid Villalba (Spain) Group Rosario Phone: oratalla Villalba Group Rosario Moratalla Villalba Group Rosario Moratalla Villalba Group Rosario Moratalla Villalba Fax: oup Rosario Moratalla Villalba Group Rosario Moratalla Villalba Group Rosario moratalla@cajal.csic.es Moratalla Villalba Group Rosario oratalla Villalba Group Rosario Moratalla Villalba Group Rosario Moratalla Villalba Group Rosario Moratalla Villalba LIST OF PERSONNEL Rosario Moratalla Villalba Principal Investigator, CSIC Professor Noelia Granado Martínez Postdoctoral Fellow Luz María Suárez González Postdoctoral Fellow Patricia García Sanz Postdoctoral Fellow José Rubén García Postdoctoral Fellow Liliana Mendieta Martínez Postdoctoral Fellow Irene Ruiz de Diego Postdoctoral Fellow Sara Ares Santos PhD student Óscar Solís Castrejón PhD student Isabel Espadas Villanueva PhD student José María Caramés PhD student Lorena Orgaz Master s student Guillermo Bueno Gil Master s student Samuel Alberquilla Master s student Karen Barraza Master s student Leticia Perote Student Horacio Javier Escobar Student María Emilia Rubio Rubio Lab Technician Beatriz Pro Lab Technician 188

195 2014 Annual Report Group Rosario Moratalla Villalba Summary During the year 2014, we study the role of the nitrergic system in L-DOPA-induced dyskinesia (LID). We assessed the effect of NO levels and NO/cyclic guanosine monophosphate (cgmp) signaling on LID in the Pitx3 (-/-) aphakia mouse, a genetic model of Parkinson s disease (PD). To evaluate the effect of decreased NO signaling on the development of LID, Pitx3 mice (-/-) were chronically treated with l-dopa and 7-nitroindazole (7-NI, a neuronal NOS inhibitor). The chronic 7-NI treatment attenuated the development of LID in the Pitx3 (-/-) mice. Moreover, the sub-acute 7-NI treatment attenuated established dyskinesia without affecting the beneficial therapeutic effect of L-DOPA. Besides, 7-NI causes a significant reduction of FosB and pach3 expression in the acutely and chronically L-DOPA-treated mice. We also examined how increasing NO/cGMP signaling affects LID by acutely administering molsidomine (an NO donor) or zaprinast (a cgmp phosphodiesterase 5-PDE5 inhibitor) before L-DOPA in mice with established dyskinesia. Paradoxically, the administration of either of these drugs also significantly diminished the expression of established LID; however, this effect occurred by expense of the antiparkinsonian L-DOPA properties. We demonstrated that targeting NO/cGMP signaling pathway is able to diminish dyskinetic movements and its molecular markers, but only the 7-NI treatment preserved the antiparkinsonian effect of L-DOPA, indicating that NOS inhibitors represent a potential therapy to reduce LID (Solis et al., 2015). Also we studied the role of DREAM (downstream regulatory element antagonist modulator) in the dyskinesia by L-DOPA. In dominant active mutant DREAM (dadream) mice, L-DOPA-induced dyskinesia was decreased throughout the entire treatment. In correlation with these behavioral results, dadream mice showed a decrease in FosB, phosphoacetylated histone H3, dynorphin-b, and phosphorylated glutamate receptor subunit, type 1 expression. Conversely, genetic inactivation of DREAM potentiated the intensity of dyskinesia, and DREAM knockout mice (-/-) exhibited an increase in expression of molecular markers associated with dyskinesia. The DREAM modifications did not affect the kinetic profile or antiparkinsonian efficacy of L-DOPA therapy. These data suggest that therapeutic approaches that activate DREAM may be useful to alleviate L-DOPA-induced dyskinesia without interfering with the therapeutic motor effects of L-DOPA (Ruiz de Diego et al., 2015). This study has been done in collaboration with Dr. Naranjo from CIBERNED and with Dr. Vallejo from CIBERDEM. Keywords Parkinson s disease, Dyskinesia, Neurotoxicity, Dopamine cell death, Addiction and role of the dopaminergic system in learning and memory processes Publications Vergano-Vera E, Diaz-Guerra E, Rodriguez-Traver E, Mendez-Gomez HR, Solis O, Pignatelli J, et al. Nurr1 blocks the mitogenic effect of FGF-2 and EGF, inducing olfactory bulb neural stem cells to adopt dopaminergic and dopaminergic-gabaergic neuronal phenotypes. Developmental neurobiology. [Epub ahead of print]. Epub 2014 Nov 28. Ruiz-DeDiego I, Mellstrom B, Vallejo M, Naranjo JR, Moratalla R. Activation of DREAM (Downstream Regulatory Element Antagonistic Modulator), a Calcium-Binding Protein, Reduces L-DOPA-Induced Dyskinesias in Mice. Biological psychiatry. 2015;77(2): Epub 2014 Mar 27. Heumann R, Moratalla R, Herrero MT, Chakrabarty K, Drucker-Colin R, Garcia-Montes JR, et al. Dyskinesia in Parkinson's disease: mechanisms and current non-pharmacological interventions. Journal of neurochemistry. 2014;130(4): Epub 2014 Jun

196 Carmena A, Granado N, Ares-Santos S, Alberquilla S, Tizabi Y, Moratalla R. Methamphetamine- Induced Toxicity in Indusium Griseum of Mice is Associated with Astro- and Microgliosis. Neurotoxicity research. [Epub ahead of print]. Epub 2014 Dec 10. Solis O, Espadas I, Del-Bel EA, Moratalla R. Nitric oxide synthase inhibition decreases l-dopa-induced dyskinesia and the expression of striatal molecular markers in Pitx3(-/-) aphakia mice. Neurobiology of disease. 2015;73: Epub 2014 Sep 30. Castro-Hernandez J, Afonso-Oramas D, Cruz-Muros I, Salas-Hernandez J, Barroso-Chinea P, Moratalla R, et al. Prolonged treatment with pramipexole promotes physical interaction of striatal dopamine D3 autoreceptors with dopamine transporters to reduce dopamine uptake. Neurobiology of disease. 2014;74C: Epub 2014 Dec 12. Villar-Cheda B, Dominguez-Meijide A, Valenzuela R, Granado N, Moratalla R, Labandeira-Garcia JL. Aging-related dysregulation of dopamine and angiotensin receptor interaction. Neurobiology of aging. 2014;35(7): Epub 2014 Jan 17. Urrutia A, Granado N, Gutierrez-Lopez MD, Moratalla R, O'Shea E, Colado MI. The JNK inhibitor, SP600125, potentiates the glial response and cell death induced by methamphetamine in the mouse striatum. The international journal of neuropsychopharmacology / official scientific journal of the Collegium Internationale Neuropsychopharmacologicum (CINP). 2014;17(2): Gonzalez-Aparicio R, Moratalla R. Oleoylethanolamide reduces L-DOPA-induced dyskinesia via TRPV1 receptor in a mouse model of Parkinson s disease. Neurobiology of disease. 2014;62: Granado N, Ares-Santos S, Moratalla R. D1 but not D4 dopamine receptors are critical for MDMAinduced neurotoxicity in mice. Neurotoxicity research. 2014;25(1): Suarez LM, Solis O, Carames JM, Taravini IR, Solis JM, Murer MG, et al. L-DOPA treatment selectively restores spine density in dopamine receptor D2-expressing projection neurons in dyskinetic mice. Biological psychiatry. 2014;75(9): Research Projects Code: PI2013/01 Title: Propiedades emergentes de la relación neurona-glía que subyacen a neurodegeneración y demencia en la enfermedad de Alzheimer Principal Investigator: Ignacio Torres-Alemán CIBERNED s collaboration: Yes Groups CIBERNED: G413; G204; G409; G108; G411; G415 Type: National Funding Agency: CIBERNED Funding: Duration: Code: Cajal Blue Brain Project Title: Cajal Blue Brain Project. International Blue Brain Proyect Principal Investigator: Javier de Felipe CIBERNED s collaboration: Yes Groups CIBERNED: G403; G204 Type: International Funding Agency: Ecole Polytechnique Federale de Lausanne (Suiza), IBM, Universidad Politécnica de Madrid, Consejo Superior de Investigaciones Científicas, España Funding: N.D. Duration: Code: PNSD Title: Caracterización de la función de DREAM en la neurotoxicidad de la metanfetamina y otros psicoestimulantes Principal Investigator: Rosario Moratalla Groups CIBERNED: Type: International Funding Agency: Ministerio Sanidad Servicios Sociales e Igualdad. Plan Nacional Sobre Drogas Funding: Duration: Code: SECITI nº 065/2013 Title: Efecto de la estimulación magnética transcraneal repetitiva (EMTr) sobre la terapia dopaminérgica y las discinesias en modelos experimentales y en pacientes con enfermedad de Parkinson avanzada Principal Investigator: Rosario Moratalla Groups CIBERNED: Program 2 190

197 2014 Annual Report Rosario Moratalla Villalba Group Type: International Funding Agency: Secretaría de Ciencia, Tecnología e Innovación del Distrito Federal, Mexico Funding: Duration: Code: S2011/BMD 2336 Title: Estudio de la biología de células madre neurales para su empleo en terapia celular en enfermedades neurodegenerativas Principal Investigator: Rosario Moratalla CIBERNED s collaboration: Yes Groups CIBERNED: G305; G204; G108 Tipo: CCAA Funding Agency: Comunidad Autónoma de Madrid Funding: Duration: Code: SAF R Title: Mecanismos moleculares responsables de la plasticidad sináptica y remodelación estructural de las neuronas estriatales de proyección en ratones disquineticos Principal Investigator: Rosario Moratalla Groups CIBERNED: Type: National Funding Agency: Ministerio de Economia y Competitividad Funding: Duration: PhD Dissertations Author: Sara Ares Santos Title: Papel de los receptores dopaminérgicos en al neurotoxicidad inducida por metanfetamina Date: 18th February 2014 Supervisor: Rosario Moratalla Villalba Author: Ana Belen Martin Title: Implicación de los receptores D2 en la respuesta conductual, celular y molecular inducida por drogas de abuso como cocaína y THC Date: 12th June 2014 Supervisor: Rosario Moratalla Villalba Author: Irene Ruiz de Diego Title: Intracellular signaling pathways involved in L-Dopa induced dyskinesias Date: 27th June 2014 Supervisor: Rosario Moratalla Villalba 191

198 Group José Ramón Naranjo Orovio Program 2 CONTACT DETAILS oup José Ramón Naranjo Orovio Group José Ramón Naranjo Orovio CSIC Group Centro Nacional José Ramón de Biotecnología Naranjo Orovio Group José món Naranjo Orovio José Ramón Naranjo Orovio Group José Ramón Naranjo Orovio Group Mol. Cell. José Biol. Ramón Naranjo ovio Group José Ramón Naranjo Orovio Group José aranjo Group José Ramón Naranjo Orovio Darwin, Group 3 José Ramón Madrid (Spain) ranjo Orovio Group José Ramón Naranjo Orovio Group José Ramón Group José Ramón Naranjo Orovio Group José Phone: món Naranjo Orovio Group José Ramón Naranjo Orovio Group José Ramón Naranjo naranjo@cnb.csic.es Orovio José Ramón Naranjo ovio Group José Ramón Naranjo Orovio Group José Ramón Naranjo Orovio Group José Ramón Naranjo LIST OF PERSONNEL Jose Ramón Naranjo Principal Investigator Britt Mellström CIBERNED Senior Researcher Juan Casado Postdoctoral Fellow JAE-DOC Alberto Rabano Researcher Javier Morón Oset Student Elena Higuera Master s student M. Paz Gonzalez CIBERNED Assistant researcher Xose Manuel Dopazo CIBERNED Assistant researcher 192

199 2014 Annual Report Group José Ramón Naranjo Orovio Summary We investigate the nuclear components of activity- and Ca2+-dependent transcriptional responses in neurons to i) understand the molecular determinants of downstream events that are responsible for plastic changes in synaptic functionality during neurodegeneration, ii) to develop tools to expose early markers of the neurodegenerative process, and iii) to design small molecules to intervene in physiological and pathological output processes including learning and memory, motor coordination and neurodegeneration. Early compensatory changes in transcriptional programs and altered neuronal calcium homeostasis are common features of many neurodegenerative pathologies including Alzheimer s (AD), Down syndrome (DS) and Huntington s disease (HD). DREAM (DRE Antagonist Modulator), a Ca2+-dependent transcriptional repressor, also known as calsenilin because its interaction with presenilins, has an important role in neurodegenerative diseases (NDD) through the control of Ca2+ homeostasis. Importantly, changes in DREAM levels are found in several NDD mouse models, including AD, DS and HD. Genetic experiments show that this could be part of a neuroprotective mechanism. In addition, we have found that DREAM is important in the development of L-DOPA-induced dyskinesias. We foresee the Ca2+-dependent transcriptional repressor DREAM as an active/central component of several different nucleoprotein complexes that mediate specifically in the various transcriptional cascades triggered by membrane depolarization in neurons that are essential in neuronal plasticity and synaptic dysfunction. Work in progress analyzes the role of DREAM in the regulation of transcription in cell and animal models of NDDs to better understand early changes in the transcriptome and epigenome and to explore new venues for therapeutic intervention oriented to boost early endogenous neuroprotective mechanisms. Furthermore, previous work in our group has shown that many biological activities of DREAM depend on specific protein-protein interactions. In this regard, we have been using NAPPA protein arrays to further expand our knowledge of the DREAM interactome. Work in progress investigates in depth some of these new interactions, especially those between DREAM and key components of the unfolded protein response pathway. Finally, we have developed in vitro and in vivo assays to screen for small molecules able to bind to and modify DREAM biological activity. The goal is to identify potent inhibitors of the DREAM activity in vivo that could represent new therapeutic venues for the treatment of NDD patients. Keywords Calcium pathologies, DREAM, Huntington s, Alzheimer s, dyskinesias, UPR, DREAM inhibitors 193

200 Publications Ruiz-DeDiego I, Mellstrom B, Vallejo M, Naranjo JR, Moratalla R. Activation of DREAM (Downstream Regulatory Element Antagonistic Modulator), a Calcium-Binding Protein, Reduces L-DOPA-Induced Dyskinesias in Mice. Biological psychiatry. 2015;77(2): Epub 2014 Mar 27. Garcia MC, Cinquina V, Palomo-Garo C, Rabano A, Fernandez-Ruiz J. Identification of CB2 receptors in human nigral neurons that degenerate in Parkinson's disease. Neuroscience letters. 2015;587:1-4. Epub 2014 Dec 3. Alonso R, Pisa D, Rabano A, Carrasco L. Alzheimer's disease and disseminated mycoses. Eur J Clin Microbiol Infect Dis. 2014;33(7): Garatachea N, Emanuele E, Calero M, Fuku N, Arai Y, Abe Y, et al. ApoE gene and exceptional longevity: Insights from three independent cohorts. Exp Gerontol. 2014;53: Pisa D, Alonso R, Juarranz A, Rabano A, Carrasco L. Direct visualization of fungal infection in brains from patients with Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2015;43(2): Epub Alonso R, Pisa D, Marina AI, Morato E, Rabano A, Carrasco L. Fungal infection in patients with Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;41(1): Sanchez-Ferro A, Rabano A, Catalan MJ, Rodriguez-Valcarcel FC, Diez SF, Herreros-Rodriguez J, et al. In vivo gastric detection of α-synuclein inclusions in Parkinson s disease. Mov Disord. 2015;30(4): Epub 2014 Aug 11. Neant I, Mellström B, Gonzalez P, Naranjo JR, Moreau M, Leclerc C. Kcnip1 a Ca2+-dependent transcriptional repressor regulates the size of the neural plate in Xenopus. Biochimica et biophysica acta. [Epub ahead of print]. Epub 2014 Dec 10. Olazaran J, Ramos A, Boyano I, Alfayate E, Valenti M, Rabano A, et al. Pattern of and risk factors for brain microbleeds in neurodegenerative dementia. Am J Alzheimers Dis Other Demen. 2014;29(3): Rabano A, Rodal I, Cuadros R, Calero M, Hernandez F, Avila J. Argyrophilic grain pathology as a natural model of tau propagation. Journal of Alzheimer's disease: JAD. 2014;40(Suppl 1):S Llorens-Martin M, Jurado-Arjona J, Fuster-Matanzo A, Hernandez F, Rabano A, Avila J. Peripherally triggered and GSK-3β-driven brain inflammation differentially skew adult hippocampal neurogenesis, behavioral pattern separation and microglial activation in response to ibuprofen. Translational psychiatry. 2014;4:e463. Epub 2014 Oct 14. Gomez-Ramos A, Sanchez-Sanchez R, Muhaisen A, Rabano A, Soriano E, Avila J. Similarities and differences between exome sequences found in a variety of tissues from the same individual. PloS one. 2014;9(7):e Epub 2014 Jul 1. Parcerisas A, Rubio SE, Muhaisen A, Gomez-Ramos A, Pujadas L, Puiggros M, et al. Somatic signature of brain-specific single nucleotide variations in sporadic Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;42(4): Llorens-Martin M, Blazquez-Llorca L, Benavides-Piccione R, Rabano A, Hernandez F, Avila J, et al. Selective alterations of neurons and circuits related to early memory loss in Alzheimer's disease. Front Neuroanat. 2014;8:38. Epub 2014 May 27. de Pedro-Cuesta J, Mahillo-Fernandez I, Calero M, Rabano A, Cruz M, Siden Å, et al.; EUROSURGYCJD Research Group. Towards an age-dependent transmission model of acquired and sporadic Creutzfeldt-Jakob disease. PloS one. 2014;9(10):e Epub 2014 Oct 3. Lastres-Becker I, Innamorato NG, Jaworski T, Rabano A, Kügler S, Van Leuven F, et al. Fractalkine activates NRF2/NFE2L2 and heme oxygenase 1 to restrain tauopathy-induced microgliosis. Brain: a journal of neurology. 2014;137(1): Mellström B, Sahun I, Ruiz-Nuno A, Murtra P, Gomez-Villafuertes R, Savignac M, et al. DREAM controls the on/off switch of specific activity-dependent transcription pathways. Molecular and cellular biology. 2014;34(5): Program 2 194

201 2014 Annual Report José Ramón Naranjo Orovio Group Research Projects Code: S2010_BMD Title: Redes de señalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas Principal Investigator: José González Castaño CIBERNED s collaboration: Yes Groups CIBERNED: G401; G104; G111; G307; G409; G412 Other CIBER s collaboration: Yes (CIBERER) Type: International Funding Agency: Comunidad de Madrid Funding: Duration: Code: 2013/07A Title: Papel de GSK-3 β en las alteraciones de los circuitos corticales que ocurren en la enfermedad de Alzheimer Principal Investigator: Teresa Iglesias Vacas CIBERNED s collaboration: Yes Groups CIBERNED: G401; G403; G111; G504; G307 Type: International Funding Agency: CIBERNED Funding: Duration:

202 Group José Ángel Obeso Inchausti Program 2 CONTACT DETAILS oup José Ángel Obeso Inchausti Group José Ángel Obeso Centro Inchausti Integral Group de Neurociencias José Ángel Obeso AC (CINAC) Inchausti Group José gel Obeso Inchausti Group José Ángel Obeso Inchausti Group José Ángel Obeso Hospital Inchausti HM Puerta Group del Sur José Ángel Obeso chausti Group José Ángel Obeso Inchausti Group José Ángel Obeso Inchausti Universidad Group San José Pablo Ángel CEUObeso Inchausti Avenida Carlos V, 70 oup José Ángel Obeso Inchausti Group José Ángel Obeso Inchausti Group José Ángel Obeso Inchausti Group José Móstoles, Madrid (Spain) gel Obeso Inchausti Group José Ángel Obeso Inchausti Group José Ángel Obeso Phone: Inchausti Group José Ángel Obeso chausti Group José Ángel Obeso Inchausti Group José Ángel Obeso jobeso.hmcinac@hmhospitales.com Inchausti Group José Ángel Obeso Inchausti G LIST OF PERSONNEL José Ángel Obeso Inchausti Principal Investigator María Ledia Fernández Hernández Research staff Inés Trigo Damas Research staff Francisco Molinet Dronda PhD student Cristina Calvo González Administrative assistant Francisco Javier Blesa de los Mozos PhD Stéphanie Etienne PhD student Raúl Martínez Fernández PhD student Ignacio Obeso Martín PhD Lydia Vela Desojo PhD Fernando Alonso Frech PhD 196

203 2014 Annual Report José Ángel Obeso Inchausti Group Summary The Movement Disorders Laboratory is directed by Dr. Obeso currently at the Centro Integral de Neurociencias AC (CINAC) in the HM Puerta del Sur in Mostoles, Madrid. The main priority of the group is to study the onset and progression of Parkinson s disease (PD) both clinically and experimentally. The group is primarily aimed to develop a therapy capable of halting PD progression. Towards this objective, we are concentrated in identify potential factors responsible for the selective vulnerability of the Substantia nigra pars compacta (SNc) and how initial compensatory mechanisms can actually facilitate disease progression. Experimental Studies Compensatory mechanisms The cardinal features of (PD) are associated with dopaminergic cell loss in the SNc. It is well demonstrated both in PD patients and animal models that parkinsonian motor features appear when SNc cell loss reaches 50-60% and striatal dopamine falls below 70-80%, suggesting some powerful compensatory mechanisms. Traditionally, the major pathophysiological emphasis in PD has resided in striatal dopamine deficit. However, the SNc has projections to other basal ganglia nuclei, and besides the thalamus, brainstem and cortex are dopaminergically innervated. To investigate the role and impact of modulating dopaminergic activity of extra-striatal basal ganglia nuclei we infuse dopaminergic agonists and antagonistswas infused locally into the somatomotor region of the subthalamic nucleus (STN) and the external segment of the globus pallidus (GPe) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys (Macaca fascicularis) at different motor states and their behavioural effects are being evaluated. Also, we have now mastered progressive models of PD in the primate (MPTP) and rat (6-OHDA) to study the metabolic pattern associated with early vs. late dopaminergic deficit, thus defining the major pathways changes in the pre-symptomatic stages. These studies involved PET imaging techniques (18F-DOPA and 11C-DTBZ) and electrophysiological, molecular and biochemical assessment of the basal ganglia and thalamus. Prion-like spreading of synuclein aggregates A study in Murcia University in collaboration with Bordeaux University and also with Prof. Miquel Vila s group in Vall d Hebron Research Institute is under way to study the factors determining the spreading of Lewy bodies (from PD brains) in normal baboons. This is aiming to obtain the first real model of PD synucleopathy in a monkey. Studies in patients with Parkinson s disease Neurophysiology of the subthalamic nucleus We finalized a study of field potentials in the STN through electrodes implanted for deep brain stimulations (DBS) and define specific oscillations associated with inhibition of an initiated task (stop-go task) or the selection of a particular action among several options. This is also accompanied by a study of fmri while performing a similar test of inhibition. Cognitive Impairment and Functional Neuroimaging Studies We have completed a longitudinal and cross-sectional study applying a neuropsychological battery of specific tests for parkinsonian patients with mild cognitive impairment and dementia, and neuroimaging based on Z-maps to compare the metabolic abnormalities observed by PET (18F-FDG) (hypometabolism) with structural anomalies objectified by morphometry MRI (atrophy). This has allowed us to define the possible anatomical and functional bases of cognitive impairment. Specifically, this study has led to propose the concept of Neurodegenerative Penumbra that will have important use and applications in the near future. 197

204 Keywords Parkinson s disease, Substantia Nigra pars compacta, Basal Ganglia, Dopamine, Vulnerability, Compensatory mechanisms, Cognitive impairment Publications Schneider SA, Obeso JA. Clinical and Pathological Features of Parkinson's Disease. Current topics in behavioral neurosciences. [Epub ahead of print]. Epub 2014 May 22. Pifl C, Rajput A, Reither H, Blesa J, Cavada C, Obeso JA, et al. Is Parkinson's disease a vesicular dopamine storage disorder? Evidence from a study in isolated synaptic vesicles of human and nonhuman primate striatum. The Journal of neuroscience: the official journal of the Society for Neuroscience. 2014;34(24): Olanow CW, Obeso J. Profile of Mahlon DeLong and Alim Benabid, 2014 Lasker-DeBakey Medical Research Awardees. Proceedings of the National Academy of Sciences of the United States of America. 2014;111(50): Epub 2014 Dec 8. Standaert DG, Olanow CW, Obeso JA. Scientific perspectives. Movement disorders: official journal of the Movement Disorder Society. 2014;29(10):1230. Olanow CW, Obeso JA. The end of the term. Movement disorders: official journal of the Movement Disorder Society. 2014;29(14): Berg D, Postuma RB, Bloem B, Chan P, Dubois B, Gasser T, et al. Time to redefine PD? Introductory statement of the MDS Task Force on the definition of Parkinson's disease. Movement disorders: official journal of the Movement Disorder Society. 2014;29(4): Epub 2014 Mar 11. Hallett M, Obeso J. Where does chorea come from? Cortical excitability findings challenge classic pathophysiological concepts. Movement disorders: official journal of the Movement Disorder Society. 2015;30(2): Epub 2014 Dec 27. Gonzalez-Redondo R, Garcia-Garcia D, Clavero P, Gasca-Salas C, Garcia-Eulate R, Zubieta JL, et al. Grey matter hypometabolism and atrophy in Parkinson's disease with cognitive impairment: a twostep process. Brain: a journal of neurology. 2014;137(Pt 8): Epub 2014 Jun 20. Gasca-Salas C, Estanga A, Clavero P, Aguilar-Palacio I, Gonzalez-Redondo R, Obeso JA, et al. Longitudinal assessment of the pattern of cognitive decline in non-demented patients with advanced Parkinson's disease. Journal of Parkinson's disease. 2014;4(4): Obeso JA, Rodriguez-Oroz MC, Stamelou M, Bhatia KP, Burn DJ. The expanding universe of disorders of the basal ganglia. Lancet. 2014;384(9942): Epub 2014 Jun 18. Obeso I, Wilkinson L, Casabona E, Speekenbrink M, Luisa Bringas M, Alvarez M, et al. The subthalamic nucleus and inhibitory control: impact of subthalamotomy in Parkinson's disease. Brain: a journal of neurology. 2014;137(Pt 5): Epub 2014 Mar 22. Marin C, Bonastre M, Mengod G, Cortes R, Giralt A, Obeso JA, et al. Early L-dopa, but not pramipexole, restores basal ganglia activity in partially 6-OHDA-lesioned rats. Neurobiology of disease. 2014;64: Toledo JB, Lopez-Azcarate J, Garcia-Garcia D, Guridi J, Valencia M, Artieda J, et al. High beta activity in the subthalamic nucleus and freezing of gait in Parkinson's disease. Neurobiology of disease. 2014;64:60-5. Recasens A, Dehay B, Bove J, Carballo-Carbajal I, Dovero S, Perez-Villalba A, et al. Lewy body extracts from Parkinson disease brains trigger α-synuclein pathology and neurodegeneration in mice and monkeys. Annals of neurology. 2014;75(3): Program 2 198

205 2014 Annual Report José Ángel Obeso Inchausti Group Research Projects Code: MJFF Staff Initiated 2013 Title: Prion-like dissemination of synuclein pathology: a non-human primate study Principal Investigator: Erwan Bezard (Universite de Bordeaux, France) CIBERNED s collaboration: Yes Groups CIBERNED: G205; G109 Type: International Funding Agency: Michael J. Fox Foundation Funding: Duration: Code: SAF C02-01 Title: Progresión del déficit dopaminérgico y mecanismos extra-estriatales en la enfermedad de Parkinson: compensación inicial vs progresión secundaria Principal Investigator: José Ángel Obeso Inchausti Groups CIBERNED: Type: International Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: 2014/06 Title: Inicio y Progresión de la Enfermedad de Parkinson: Papel de la Activaciñon Glial Principal Investigator: María Cruz Rodríguez Oroz CIBERNED s collaboration: Yes Groups CIBERNED: G206; G404; G205 Type: International Funding Agency: N.D. Funding: Duration:

206 Group Ana María Pérez Castillo Program 2 CONTACT DETAILS oup Ana María Pérez Castillo Group Ana María Pérez Instituto Castillo de Group Investigaciones Ana María Biomédicas Pérez Castillo (CSIC-UAM) Group Ana María Pérez LIST OF PERSONNEL stillo Group Ana María Pérez Castillo Group Dpto. Ana María Modelos Pérez Experimentales Castillo Group de Enfermedades Ana María Pérez Humanas Castillo Group Ana aría Pérez Castillo Group Ana María Pérez Castillo Group Ana María Pérez Castillo Arturo Group Duperier, Ana María 4 Pérez Castillo Madrid (Spain) oup Ana María Pérez Castillo Group Ana María Pérez Castillo Group Ana María Pérez Castillo Group Ana María Phone: rez Castillo Group Ana María Pérez Castillo Group Ana María Pérez Castillo Group Fax: Ana +34 María Pérez 401Castillo Group a María Pérez Castillo Group Ana María Pérez Castillo Group Ana María Pérez Castillo aperez@iib.uam.es Group Ana María Pérez LIST OF PERSONNEL Ana María Pérez Castillo Principal Investigator, CSIC Professor Angel Santos Montes Full Professor Elena Giné Postdoctoral Fellow José A. Morales García Postdoctoral Fellow Elena Hernández Encinas Postdoctoral Fellow Marina Sanz SanCristóbal Technician Sandra Alonso Gil Technician David Martínez Pablo Student Ana Tamarit Student 200

207 2014 Annual Report Ana María Pérez Castillo Group Summary During the last year we have analyzed the effects of different inhibitors of GSK3 and PDE7 enzymes on neurodegenerative diseases, mainly Parkinson s disease, and neurogenesis. Specifically, we have investigated the effects of the PDE7 inhibitor S14, on adult neurogenesis in the 6-OHDA model of PD mimicking a severe dopaminergic striatal deficit. We have demonstrated that pharmacological manipulation of PDE7 in vivo induces a strong neurogenesis in the SNpc of 6-OHDA-lesioned animals towards a dopaminergic phenotype. In vitro, PDE7 inhibition increased the neuronal differentiation of neurospheres obtained either from embryonic ventral midbrain or adult SVZ. Then, by using PDE7 inhibitors, we could potentially contribute to up-regulate endogenous neurogenesis, and/or favor integration of new dopaminergic neurons to stimulate neurorepair in PD. Regarding the role of GSK3 inhibition on neurogenesis we have shown that inhibition of GSK3 induces proliferation, migration, and differentiation of neural stem cells towards a neuronal phenotype in in vitro studies. Also, we have demonstrated that inhibition of GSK3 with the small molecule NP03112, called tideglusib, induces neurogenesis in the dentate gyrus of the hippocampus of adult rats. On the other hand, we are collaborating with Dr. García-Verdugo s group analyzing the effects of ligands of the nuclear receptor PPARg upon neurogenesis and oligodendrogenesis during aging. Additionally, we have continued studying how PDE7 inhibition affects neuroprotection and neuroinflammarion processes during PD development. Previous studies from our group have shown that small chemical inhibitors of this enzyme are potent neuroprotective agents of dopaminergic neurons after lipopolysaccharide or 6-OHDA injuries. Very recently, we have shown that lentiviral-mediated delivery of PDE7B shrna conferred marked in vitro and in vivo neuroprotection against 6-OHDA and LPS toxicity in dopaminergic neurons, and preserved motor function involving the dopamine system in mouse. Our results substantiate our previous data and provide a validation of PDE7B enzyme as a valuable new target for therapeutic development in the treatment of PD. Keywords GSK-3, Neuroprotection, Neuroinflammation, Neurogenesis, Parkinson, PDE7, PPARg Publications Morales-Garcia JA, Palomo V, Redondo M, Alonso-Gil S, Gil C, Martinez A, et al. Crosstalk between phosphodiesterase 7 and glycogen synthase kinase-3: two relevant therapeutic targets for neurological disorders. ACS Chemical Neuroscience. 2014;5(3): Epub 2014 Jan 17. Garcia AM, Brea J, Morales-Garcia JA, Perez DI, Gonzalez A, Alonso-Gil S, et al. Modulation of campspecific PDE without emetogenic activity: new sulfide-like PDE7 inhibitors. Journal of medicinal chemistry. 2014;57(20): Epub 2014 Oct 8. Prati F, De Simone A, Bisignano P, Armirotti A, Summa M, Pizzirani D, et al. Multitarget Drug Discovery for Alzheimer's Disease: Triazinones as BACE-1 and GSK-3β Inhibitors. Angewandte Chemie (International ed. in English). 2015;54(5): Epub 2014 Dec 11. Lopez-Iglesias B, Perez C, Morales-Garcia JA, Alonso-Gil S, Perez-Castillo A, Romero A, et al. New melatonin-n,n-dibenzyl(n-methyl)amine hybrids: potent neurogenic agents with antioxidant, cholinergic, and neuroprotective properties as innovative drugs for Alzheimer's disease. Journal of medicinal chemistry. 2014;57(9): Epub 2014 Apr

208 Morales-Garcia JA, Aguilar-Morante D, Hernandez-Encinas E, Alonso-Gil S, Gil C, Martinez A, et al. Silencing phosphodiesterase 7B gene by lentiviral-shrna interference attenuates neurodegeneration and motor deficits in hemiparkinsonian mice. Neurobiology of aging. 2015;36: Epub 2014 Oct 13. Research Projects Code: SAF Title: Assessment of novel PPARγ activators and GSK-3β and PDE7 inhibitors as possible therapeutic agents for the treatment of Parkinson Disease and other neurodegenerative disorders Principal Investigator: Ana Pérez Castillo Groups CIBERNED: Type: National Funding Agency: MICINN Funding: Duration: Code: IPT Title: Desarrollo de terapias innovadoras para la enfermedad de Parkinson Principal Investigator: Araclon Biotech Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: 2014 Code: Functional validation in animal and cellular models of genetic determinants of diseases and ageing processes Title: Identification of novel pharmacotherapy in epilepsy disorders. A multimodal investigation (Patho-physiology and therapy of epilepsy and epileptiform disorders Principal Investigator: Ana Pérez Castillo Groups CIBERNED: Type: International Funding Agency: Unión Europea Funding: N.D. Duration: 2014 Code: Marie Skłodowska-Curie Actions. H2020-MSCA- ITN-2014 Title: Training and Research in epilepsy disease Principal Investigator: Ana Pérez Castillo Groups CIBERNED: Type: International Funding Agency: Unión Europea Funding: N.D. Duration: 2014 Code: INNOGRAPH H2020-PHC-2014-two-stage Title: INNOvative in vitro Point-of-Care GRAPHene Technologies for Clinical Applications in Neurodegenerative Diseases Principal Investigator: Emmanuel Ifeachor Groups CIBERNED: Type: International Funding Agency: Unión Europea Funding: N.D. Duration: Program 2 202

209 2014 Annual Report Ana María Pérez Castillo Group PhD Dissertations Author: Elena Hernández Encinas Title: Papel del Componente 3 del Complemento (C3) como mediador de C/EBPbeta en los procesos inflamatorios y excitotóxicos del Sistema Nervioso Central Date: 28th March 2014 Supervisor: Ana María Pérez Castillo 203

210 Group Jordi Pérez Tur Program 2 CONTACT DETAILS oup Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Unitat Jordi de Pérez Genètica Tur Group Molecular Jordi Pérez Tur rdi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Institut de Jordi Biomedicina Pérez Tur Group de València-CSIC Jordi Pérez Tur Group Jaume Roig, 11 ez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Tur Valencia (Spain) oup Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Phone: Jordi Pérez Tur Group Jordi Pérez Tur Fax: oup Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur di jpereztur@ibv.csic.es rez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pérez Tur Group Jordi Pé LIST OF PERSONNEL Jordi Pérez-Tur Principal Investigator Fernando Cardona Serrate Advanced Technical Irene Nebot Ruiz FPU Researcher Concepción Rubio Granero Junior researcher 204

211 2014 Annual Report Jordi Pérez Tur Group Summary This year, our group has deepened, through collaboration with other groups CIBERNED in the genetic basis of neurodegenerative diseases. Basically we worked on defining the impact of variations in risk conferred by TREMH1 against Alzheimer s disease. Moreover we have also participated in characterizing an abnormal expansion in Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia patients in the C9ORF72 locus. Finally, we have determined the existence of a founder effect in samples of carbomyl phosphate sintase 1 (CPS1) defficiency patients of Turkish origin. As for our work in progress, throughout 2014 we continued the study of the genetic factors involved in the onset of dementia in Parkinson s disease, in characterizing variations or alterations in DNA related to the appearance of ALS and in the definition of the mutation that causes a familial form of Progressive Supranuclear Palsy. Keywords Alzheimer, Parkinson, Neurodegeneration, Neurogenetics, ALS Publications Hu L, Diez-Fernandez C, Rüfenacht V, Hismi BÖ, Ünal Ö, Soyucen E, et al. Recurrence of carbamoyl phosphate synthetase 1 (CPS1) deficiency in Turkish patients: characterization of a founder mutation by use of recombinant CPS1 from insect cells expression. Molecular genetics and metabolism. 2014;113(4): Epub 2014 Oct 7. Cardona F, Tormos-Perez M, Perez-Tur J. Structural and functional in silico analysis of LRRK2 missense substitutions. Mol Biol Rep. 2014;41: Epub 2014 Feb 2. Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodriguez-Rodriguez E, Lopez de Munain A, et al. Assessing the role of the TREM2 p.r47h variant as a risk factor for Alzheimer's disease and frontotemporal dementia. Neurobiology of aging. 2014;35(2):444.e1-4. PhD Dissertations Author: Luis Navarro Sánchez Title: Genetics and Epigenetics in Parkinson's Disease Date: 20th June 2014 Supervisor: Jordi Pérez Tur 205

212 Group Manuel Rodríguez Díaz Program 2 CONTACT DETAILS oup Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Departamento Rodríguez Díaz de Group Fisiología Manuel Rodríguez az Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Facultad Rodríguez de Medicina Díaz Group Manuel dríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Universidad Manuel de La Rodríguez Laguna Díaz Group anuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz La Group Laguna Manuel (Spain) Rodríguez Díaz Phone: oup Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez Fax: az Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez mrdiaz@ull.es Díaz Group Manuel dríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group Manuel Rodríguez Díaz Group LIST OF PERSONNEL Manuel Rodríguez Díaz Principal Investigator, Full Professor Tomás González-Hernández Full Professor, Researcher Magdalena Sabaté Bel Associate Professor, Researcher Ingrid Morales Pérez Researcher CIBERNED Alberto Sánchez Fernández Researcher Catalina Llanos Mendoza Fellow Domingo Alfonso Oramas Associate Researcher Ana Manzanedo López Associate Researcher Ángel Fuentes Rodríguez Technician Javier Castro Hernández Technician Josmar Salas-Hernández Technician Sergio González Martín-Fernández Technician 206

213 2014 Annual Report Manuel Rodríguez Díaz Group Summary An animal model to study the main axonal and glial mechanisms involved in the retrograde degeneration of dopaminergic neurons which is characteristic of Parkinson s disease was developed. This model is being used to analyze the role of the striatal astrocytes on the dopaminergic degeneration, paying particular attention to studying the action of the astrocytic repopulation of the striatum by cells arriving from the subventricular region on the dopamine cell degeneration. We are studying the proliferation, migration and differentiation patterns of cells arriving at the striatum from the subventricular zone, and the influence of striatal dopaminergic denervation on these variables. We are beginning to study the action of astrocytes generated from IPs cells obtained from parkinsonian patients as a possible activator vs. inhibitor of retrograde dopaminergic degeneration. Our group is collaborating in the IPs project with other groups belonging to CIBERNED who facilitate patient samples and the differentiation of astrocytic cells. A procedure to identify the main basal ganglia in magnetic resonance images, and to obtain BOLD signals which may represent the functional activity of each center was developed. The functional connectivity of basal ganglia was studied by analyzing the BOLD activity of these centers with different methods (partial correlation, main components...). The results show that the closed-loop cortico-subcortical activity found in these center in animals is also present in humans, thereby supporting the classical basal ganglia model. The power of the functional connectivity between different basal ganglia changed with the motor behaviour. Functional connectivity was also used to study the influence of the intralaminar thalamic centers on the main basal ganglia circuits. The membrane transporter involved in the dopamine uptake by dopaminergic neurons is a relevant factor for the vulnerability of mesencephalic dopaminergic neurons in Parkinson s disease. We have studied the possible role of D3 dopamine receptors on the dopamine transporter activity in mice and cell cultures. D3 agonists decrease dopamine uptake, changing the integration of the transporter in the cytoplasmatic membrane of the dopaminergic synaptic terminals, and the phosforylation and ubiquitinization of the dopamine transporter. Keywords Astrocytes, IPs, Subventricular zone, Dopaminergic degeneration, Dopamine transporter, Dopaminergic vulnerability, Functional neuroimaging Publications Castro-Hernandez J, Afonso-Oramas D, Cruz-Muros I, Salas-Hernandez J, Barroso-Chinea P, Moratalla R, et al. Prolonged treatment with pramipexole promotes physical interaction of striatal dopamine D3 autoreceptors with dopamine transporters to reduce dopamine uptake. Neurobiology of disease. 2014;74C: Epub 2014 Dec 12. Rodriguez M, Morales I, Rodriguez-Sabate C, Sanchez A, Castro R, Brito JM, et al. The degeneration and replacement of dopamine cells in Parkinson's disease: the role of aging. Frontiers in neuroanatomy. 2014;8:80. Epub 2014 Aug 7. Rodriguez-Sabate C, Llanos C, Morales I, Garcia-Alvarez R, Sabate M, Rodriguez M. The functional connectivity of intralaminar thalamic nuclei in the human basal ganglia. Human brain mapping. [Epub ahead of print]. Epub 2014 Nov

214 Research Projects Code: 59/2011 Title: Cambios cognitivos en el envejecimiento normal: un estudio de seguimiento Principal Investigator: Antonieta Nieto Barco Groups CIBERNED: Type: National Funding Agency: Ministerio de Sanidad, Seguros Sociales e Igualdad; Research Projects científica, desarrollo e innovación tecnológica Funding: Duration: Code: PSI Title: Cerebelo y Cognición: Deterioro neuropsicológico en la Ataxia de Friedreich y su relación con la degeneración cerebelosa en imagenes de RM Principal Investigator: Antonieta Nieto Barco Groups CIBERNED: Type: National Funding Agency: Ministerio de Ciencia e Innovación. Research Projects Fundamental no orientada Funding: Duration: Code: 2014/06 Title: Inicio y Progresión de la Enfermedad de Parkinson: Papel de la Activación Glial Principal Investigator: María Cruz Rodríguez Oroz CIBERNED s collaboration: Yes Groups CIBERNED: G206; G404; G205 Type: International Funding Agency: N.D. Funding: Duration: Program 2 208

215 Group Eduard Tolosa Sarró Program 2 CONTACT DETAILS Group Eduard Tolosa Sarró Group Eduard Tolosa Hospital Sarró Group Clinic de Eduard Barcelona Tolosa / Universidad Sarró Group de Eduard Barcelona Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Villarroel, Tolosa Sarró 171 Group Eduar Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Barcelona Sarró (Spain) Group Eduard Tolos Phone: Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Fax: Sarró Group Eduard Tolosa Sarr Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró etolosa@clinic.ub.es Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduard Tolosa Sarró Group Eduar LIST OF PERSONNEL Eduard Tolosa Sarró Principal Investigator, Full Professor María José Martí Doménech Consultant Francesc Valldeoriola i Serra Consultant Esteban Muñoz Martínez Consultant Joan Santamaria Cano Consultant Josep Valls-Solé Consultant Alejandro Iranzo de Riquer Consultant Carles Gaig Specialist Mario Ezquerra Trabalón Researcher Yaroslau Compta Hinrjy Specialist Rubén Fernández-Santiago Researcher Mónica Serradell Eroles PhD student Carme Junqué Plaja Full Professor Claustre Pont PhD student Dolores Vilas PhD student Judith Navarro PhD student Mercè Bonastre García Lab Technician Manuel Fernández Sánchez Lab Technician 209

216 Summary In a study in subjects with Parkinson who were brain donors we have shown deposits of synuclein aggregated in multiple peripheral organs and rostrocaudal involvement in the gastrointestinal tract (Mov Disord 2014). These studies provide a basis for the identification of peripheral synuclein as a diagnostic marker of Parkinson s. Following up on research in Parkinson s preclinical stage, we have confirmed in a series to 173 cases that most convert to a synucleinopathy (PLoS One 2014) and autopsy studies have confirmed that most suffered from synucleinopathy (Lancet Neurology 2013). We have reported that hyposmia and transcranial sonography of the substantia nigra are not proper markers of progression in the pre-motor disease (Mov Disord 2014). Within the study ONSET PD we have reported that non-motor symptoms are common in untreated early Parkinson, appearing in pre-motor stages in a majority of cases. Early motor symptoms do not allow to discriminate between patients and controls (Mov Disord 2014). Following studies on Parkinson associated to LRRK2 mutations we have shown that neuropsychiatric, dysautonomy and sleep dysfunctions are as frequent as in idiopathic Parkinson s but not the hyposmia (PLoS One 2014). In LRR2-PD hyposmia was more common in subjects with abnormal cardiac MIBG, indicative of dysautonomia (J Neurol 2011). We have continued with activity on the biomarkers research line with studies of alpha-synuclein on cerebrospinal fluid and PET-FDDNP as a predictor of dementia in Parkinson s as well as a differential biomarker in atypical and unclassifiable parkinsonism [PET-FDDNP and in vivo diagnosis of tauopathy in unclassifiable parkinsonism PI11 / (ending recruitment)]. Regarding the research line on structural and functional brain abnormalities associated with cognitive impairment in Parkinson s, we have identified a pattern of reduced cortical thickness in patients with mild cognitive impairment (MCI) located essentially on the temporal-parietal regions and correlating with alterations in semantic fluency and MMSE pentagon execution (Parkinsonism Relat Disord 2014), both predictors of progression to dementia (Mov Disord 2014). The study of functional connectivity in resting state shows reduction in long distance connections and increased local connections (Hum Brain Mapp 2014). These data suggest a reorganization in brain networks that could explains the MCI. Blood-circulating micrornas have been proposed as biomarkers in neurodegenerative diseases such as Parkinson s disease. We analyzed 377 micrornas in serum samples from idiopathic patients, patients carrying the G2019S mutation in the LRRK2 gene and healthy subjects. We found three mirnas with low expression in patients with genetic and idiopathic Parkinson. Our results indicate that the low expression of specific mirnas is associated with Parkinson s disease and suggest its potential use as noninvasive markers of disease (J Neurosci Res 2014). We then studied the expression of mir-19b, mir-29a, and mir-29c in sera from idiopathic patients with REM behavior disorder (RBD), before and after conversion into a synucleinopathy. The results are currently being analyzed. Studies in parkin knockout mouse brains and human fibroblasts from PD patients with parkin mutations, we found high levels of parkin. We also found that parkin protects neurons from cell death induced by RTP801 through its degradation, whereas knocking down parkin exacerbates neuronal death. RTP801 is a new substrate of Parkin which can contribute to neurodegeneration caused by parkin loss of function (Cell Death Dis 2014). Other relevant studies include TAUROS (Mov Disord 2014), the first clinical trial in a tauopathy with an inhibitor of GSK-3 and a pioneering study on handicap in advanced Parkinson (Europ Neurol 2014). Program 2 210

217 2014 Annual Report Eduard Tolosa Sarró Group Keywords Neuroimaging and cognition, Biological, Diagnostic pre-motor, Non-motor symptoms, Motor complications, Genetic variants, Sleep disturbances, Experimental therapy Publications Rusz J, Megrelishvili M, Bonnet C, Okujava M, Brožova H, Khatiashvili I, et al. A distinct variant of mixed dysarthria reflects parkinsonism and dystonia due to ephedrone abuse. Journal of neural transmission. 2014;121(6): Epub 2014 Jan 21. Sabater L, Gaig C, Gelpi E, Bataller L, Lewerenz J, Torres-Vega E, et al. A novel non-rapid-eye movement and rapid-eye-movement parasomnia with sleep breathing disorder associated with antibodies to IgLON5: a case series, characterisation of the antigen, and post-mortem study. Lancet neurology. 2014;13(6): Epub 2014 Apr 3. Quadri M, Yang X, Cossu G, Olgiati S, Saddi VM, Breedveld GJ, et al. An exome study of Parkinson's disease in Sardinia, a Mediterranean genetic isolate. Neurogenetics. 2015;16(1): Epub 2014 Oct 8. Proukakis C, Shoaee M, Morris J, Brier T, Kara E, Sheerin UM, et al. Analysis of Parkinson's disease brain-derived DNA for alpha-synuclein coding somatic mutations. Movement disorders: official journal of the Movement Disorder Society. 2014;29(8): Epub 2014 Apr 21. Ferini-Strambi L, Oertel W, Dauvilliers Y, Postuma RB, Marelli S, Iranzo A, et al. Autonomic symptoms in idiopathic REM behavior disorder: a multicentre case-control study. Journal of neurology. 2014;261(6): Epub 2014 Apr 1. Antonell A, Mansilla A, Rami L, Llado A, Iranzo A, Olives J, et al. Cerebrospinal fluid level of YKL- 40 protein in preclinical and prodromal Alzheimer's disease. Journal of Alzheimer's disease: JAD. 2014;42(3): Sala-Llonch R, Junque C, Arenaza-Urquijo EM, Vidal-Pineiro D, Valls-Pedret C, Palacios EM, et al. Changes in whole-brain functional networks and memory performance in aging. Neurobiology of aging. 2014;35(10): Epub 2014 Apr 19. Baggio HC, Segura B, Sala-Llonch R, Marti MJ, Valldeoriola F, Compta Y, et al. Cognitive impairment and resting-state network connectivity in Parkinson's disease. Human brain mapping. 2015;36(1): Epub 2014 Aug 28. Frauscher B, Jennum P, Ju YE, Postuma RB, Arnulf I, Cochen De Cock V, et al. Comorbidity and medication in REM sleep behavior disorder: a multicenter case-control study. Neurology. 2014;82(12): Epub 2014 Feb 19. Llull L, la Puma D, Falgàs N, Renu A, Iranzo A. Compressive C5-C6 radiculopathy secondary to spontaneous vertebral artery dissection. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Jul 4. Melia U, Guaita M, Vallverdu M, Montserrat JM, Vilaseca I, Salamero M, et al. Correntropy measures to detect daytime sleepiness from EEG signals. Physiological measurement. 2014;35(10): Epub 2014 Sep 19. Segura B, Baggio HC, Marti MJ, Valldeoriola F, Compta Y, Garcia-Diaz AI, et al. Cortical thinning associated with mild cognitive impairment in Parkinson's disease. Movement disorders: official journal of the Movement Disorder Society. 2014;29(12): Epub 2014 Aug 7. Navarro-Otano J, Casanova-Mollà J, Morales M, Valls-Sole J, Tolosa E. Cutaneous autonomic denervation in Parkinson's disease. Journal of neural transmission. [Epub ahead of print]. Epub 2014 Dec 24. Vidal-Pineiro D, Valls-Pedret C, Fernandez-Cabello S, Arenaza-Urquijo EM, Sala-Llonch R, Solana E, 211

218 et al. Decreased Default Mode Network connectivity correlates with age-associated structural and cognitive changes. Frontiers in aging neuroscience. 2014;6:256. Epub 2014 Sep 25. Cabib C, Llufriu S, Casanova-Mollà J, Saiz A, Valls-Sole J. Defective sensorimotor integration in preparation for reaction time tasks in patients with multiple sclerosis. Journal of neurophysiology. 2014;:jn Epub 2014 Dec 4. Coelho M, Marti MJ, Sampaio C, Ferreira JJ, Valldeoriola F, Rosa MM, et al. Dementia and severity of parkinsonism determines the handicap of patients in late-stage Parkinson's disease: the Barcelona- Lisbon cohort. European journal of neurology: the official journal of the European Federation of Neurological Societies. 2015;22(2): Epub 2014 Sep 15. Tafti M, Hor H, Dauvilliers Y, Lammers GJ, Overeem S, Mayer G, et al. DQB1 locus alone explains most of the risk and protection in narcolepsy with cataplexy in Europe. Sleep. 2014;37(1): Epub 2014 Jan 1. Ferini-Strambi L, Oertel W, Dauvilliers Y, Postuma RB, Marelli S, Iranzo A, et al. Erratum to: Autonomic symptoms in idiopathic REM behavior disorder: a multicentre case-control study. Journal of neurology. 2015;262(1): Epub Iranzo A, Stockner H, Serradell M, Seppi K, Valldeoriola F, Frauscher B, et al. Five-year follow-up of substantia nigra echogenicity in idiopathic REM sleep behavior disorder. Movement disorders: official journal of the Movement Disorder Society. 2014;29(14): Epub 2014 Nov 10. Lees AJ, Tolosa E, Olanow CW. Four pioneers of L-dopa treatment: Arvid Carlsson, Oleh Hornykiewicz, George Cotzias, and Melvin Yahr. Movement disorders: official journal of the Movement Disorder Society. 2015;30(1): Epub 2014 Dec 8. Baggio HC, Sala-Llonch R, Segura B, Marti MJ, Valldeoriola F, Compta Y, et al. Functional brain networks and cognitive deficits in Parkinson's disease. Human brain mapping. 2014;35(9): Epub 2014 Mar 17. Ros C, Tercero A, Alobid I, Balasch J, Santamaria J, Mullol J, et al. Hearing loss in adult women with Turner's syndrome and other congenital hypogonadisms. Gynecological endocrinology: the official journal of the International Society of Gynecological Endocrinology. 2014;30(2): Epub 2013 Nov 20. Pujol N, Penades R, Junque C, Dinov I, Fu CH, Catalan R, et al. Hippocampal abnormalities and age in chronic schizophrenia: morphometric study across the adult lifespan. The British journal of psychiatry: the journal of mental science. 2014;205(5): Epub 2014 Sep 11. Botta-Orfila T, Morato X, Compta Y, Lozano JJ, Falgàs N, Valldeoriola F, et al. Identification of blood serum micro-rnas associated with idiopathic and LRRK2 Parkinson's disease. Journal of neuroscience research. 2014;92(8): Epub 2014 Mar 20. O'Brien JT, Oertel WH, McKeith IG, Grosset DG, Walker Z, Tatsch K, et al. Is ioflupane I123 injection diagnostically effective in patients with movement disorders and dementia? Pooled analysis of four clinical trials. BMJ open. 2014;4(7):e Epub 2014 Jul 3. Alcalay RN, Aasly J, Berg D, Bressman S, Brice A, Brockmann K, et al. Michael J. Fox Foundation LRRK2 Consortium: geographical differences in returning genetic research data to study participants. Genetics in medicine: official journal of the American College of Medical Genetics. 2014;16(8): Gelpi E, Navarro-Otano J, Tolosa E, Gaig C, Compta Y, Rey MJ, et al. Multiple organ involvement by alpha-synuclein pathology in Lewy body disorders. Movement disorders: official journal of the Movement Disorder Society. 2014;29(8): Epub 2014 Jan 2. Iranzo A, Fernandez-Arcos A, Tolosa E, Serradell M, Molinuevo JL, Valldeoriola F, et al. Neurodegenerative disorder risk in idiopathic REM sleep behavior disorder: study in 174 patients. PloS one. 2014;9(2):e Epub 2014 Feb 26. Iranzo A, Gelpi E, Tolosa E, Molinuevo JL, Serradell M, Gaig C, et al. Neuropathology of prodromal Lewy body disease. Movement disorders: official journal of the Movement Disorder Society. 2014;29(3): Epub 2014 Jan 31. Sansa G, Falup-Pecurariu C, Salamero M, Iranzo A, Santamaria J. Non-random temporal distribution of sleep onset REM periods in the MSLT in narcolepsy. Journal of the neurological sciences. 2014;341(1-2): Epub 2014 Mar 26. Program 2 212

219 2014 Annual Report Eduard Tolosa Sarró Group Hoggart CJ, Venturini G, Mangino M, Gomez F, Ascari G, Zhao JH, et al. Novel approach identifies SNPs in SLC2A10 and KCNK9 with evidence for parent-of-origin effect on body mass index. PLoS genetics. 2014;10(7):e Epub 2014 Jul 31. Fagundo AB, Via E, Sanchez I, Jimenez-Murcia S, Forcano L, Soriano-Mas C, et al. Physiological and brain activity after a combined cognitive behavioral treatment plus video game therapy for emotional regulation in bulimia nervosa: a case report. Journal of medical Internet research. 2014;16(8):e183. Epub 2014 Aug 12. Kumru H, Portell E, Barrio M, Santamaria J. Restless legs syndrome in patients with sequelae of poliomyelitis. Parkinsonism & related disorders. 2014;20(10): Epub 2014 Jun 26. Grosset DG, Tatsch K, Oertel WH, Tolosa E, Bajaj N, Kupsch A, et al. Safety Analysis of 10 Clinical Trials and for 13 Years After First Approval of Ioflupane 123I Injection (DaTscan). Journal of nuclear medicine: official publication, Society of Nuclear Medicine. 2014;55(8): Epub 2014 Jun 19. Lecomte MJ, Bertolus C, Santamaria J, Bauchet AL, Herbin M, Saurini F, et al. Selective disruption of acetylcholine synthesis in subsets of motor neurons: a new model of late-onset motor neuron disease. Neurobiology of disease. 2014;65: Epub 2014 Jan 28. Santamaria J, Iranzo A. Sleep disorders matter in neurology. Lancet neurology. 2014;13(1): Gaig C, Iranzo A, Tercero A, Herman ST, Santamaria J. Stimulus-induced generalized epileptiform discharges: an unrecognized EEG pattern in refractory nonconvulsive status epilepticus. Journal of clinical neurophysiology: official publication of the American Electroencephalographic Society. 2014;31(6): Garcia-Diaz AI,Segura B,Baggio HC,Marti MJ,Valldeoriola F,Compta Y et al. Structural MRI correlates of the MMSE and pentagon copying test in Parkinson's disease. Parkinsonism & related disorders. 2014;20(12): Epub 2014 Oct 19. Jankovic J, Berkovich E, Eyal E, Tolosa E. Symptomatic efficacy of rasagiline monotherapy in early Parkinson's disease: post-hoc analyses from the ADAGIO trial. Parkinsonism & related disorders. 2014;20(6): Epub 2014 Mar 5. Marques-Iturria I, Garolera M, Pueyo R, Segura B, Hernan I, Garcia-Garcia I, et al. The interaction effect between BDNF val66met polymorphism and obesity on executive functions and frontal structure. American journal of medical genetics. Part B, Neuropsychiatric genetics: the official publication of the International Society of Psychiatric Genetics. 2014;165B(3): Epub 2014 Mar 12. Respondek G, Stamelou M, Kurz C, Ferguson LW, Rajput A, Chiu WZ, et al. The phenotypic spectrum of progressive supranuclear palsy: A retrospective multicenter study of 100 definite cases. Movement disorders: official journal of the Movement Disorder Society. 2014;29(14): Epub 2014 Nov 5. Romani-Aumedes J, Canal M, Martin-Flores N, Sun X, Perez-Fernandez V, Wewering S, et al. Parkin loss of function contributes to RTP801 elevation and neurodegeneration in Parkinson's disease. Cell death & disease. 2014;5:e1364. Epub 2014 Aug 7. Compta Y, Valente T, Saura J, Segura B, Iranzo A, Serradell M, et al. Correlates of cerebrospinal fluid levels of oligomeric- and total-α-synuclein in premotor, motor and dementia stages of Parkinson's disease. Journal of neurology. 2014;262(2): Epub 2014 Nov 8. Bras J, Guerreiro R, Darwent L, Parkkinen L, Ansorge O, Escott-Price V, et al. Genetic analysis implicates APOE, SNCA and suggests lysosomal dysfunction in the etiology of dementia with Lewy bodies. Human molecular genetics. 2014;23(23): Epub 2014 Jun 27. Sharma M, Wenning G, Krüger R; European Multiple-System Atrophy Study Group (EMSA-SG). Mutant COQ2 in multiple-system atrophy. The New England journal of medicine. 2014;371(1):80-1. Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agundez JA, Jimenez-Jimenez FJ, Ross OA, et al. TREM2 R47H variant and risk of essential tremor: A cross-sectional international multicenter study. Parkinsonism & related disorders. [Epub ahead of print]. Epub 2014 Dec 24. Gaig C, Vilas D, Infante J, Sierra M, Garcia-Gorostiaga I, Buongiorno M, et al. Nonmotor symptoms in LRRK2 G2019S associated Parkinson's disease. PloS one. 2014;9(10):e Epub 2014 Oct 17. Pont-Sunyer C, Hotter A, Gaig C, Seppi K, Compta Y, Katzenschlager R, et al. The Onset of Nonmotor Symptoms in Parkinson's disease (The ONSET PD Study). Movement disorders: official journal of the Movement Disorder Society. 2015;30(2): Epub 2014 Dec

220 Tolosa E, Litvan I, Höglinger GU, Burn D, Lees A, Andres MV, et al. A phase 2 trial of the GSK-3 inhibitor tideglusib in progressive supranuclear palsy. Movement disorders: official journal of the Movement Disorder Society. 2014;29(4): Epub 2014 Feb 14. Kalia LV, Lang AE, Hazrati LN, Fujioka S, Wszolek ZK, Dickson DW, et al. Clinical Correlations With Lewy Body Pathology in LRRK2-Related Parkinson Disease. JAMA neurology. 2015;72(1): Epub Arbizu J, Luquin MR, Abella J, de la Fuente-Fernandez R, Fernandez-Torron R, Garcia-Solis D, et al. Functional neuroimaging in the diagnosis of patients with Parkinsonism: Update and recommendations for clinical use. Revista espanola de medicina nuclear e imagen molecular. 2014;33(4): Epub 2014 Apr 14. Geurtsen GJ, Hoogland J, Goldman JG, Schmand BA, Tröster AI, Burn DJ, et al. Parkinson's disease mild cognitive impairment: application and validation of the criteria. Journal of Parkinson's disease. 2014;4(2): Höglinger GU, Huppertz HJ, Wagenpfeil S, Andres MV, Belloch V, Leon T, et al. Tideglusib reduces progression of brain atrophy in progressive supranuclear palsy in a randomized trial. Movement disorders: official journal of the Movement Disorder Society. 2014;29(4): Epub 2014 Jan 31. Tolosa E, Hernandez B, Linazasoro G, Lopez-Lozano JJ, Mir P, Marey J, et al. Efficacy of levodopa/ carbidopa/entacapone versus levodopa/carbidopa in patients with early Parkinson's disease experiencing mild wearing-off: a randomised, double-blind trial. Journal of neural transmission. 2014;121(4): Pereira JB, Valls-Pedret C, Ros E, Palacios E, Falcon C, Bargallo N, et al. Regional vulnerability of hippocampal subfields to aging measured by structural and diffusion MRI. Hippocampus. 2014;24(4): Arino H, Iranzo A, Gaig C, Santamaria J. Sexsomnia: parasomnia associated with sexual behaviour during sleep. Neurologia (Barcelona, Spain). 2014;29(3): Navarro-Otano J, Gaig C, Muxi A, Lomena F, Compta Y, Buongiorno MT, et al. 123I-MIBG cardiac uptake, smell identification and 123I-FP-CIT SPECT in the differential diagnosis between vascular parkinsonism and Parkinson's disease. Parkinsonism & related disorders. 2014;20(2): Padilla N, Junque C, Figueras F, Sanz-Cortes M, Bargallo N, Arranz A, et al. Differential vulnerability of gray matter and white matter to intrauterine growth restriction in preterm infants at 12 months corrected age. Brain research. 2014;1545:1-11. Eggington S, Valldeoriola F, Chaudhuri KR, Ashkan K, Annoni E, Deuschl G. The cost-effectiveness of deep brain stimulation in combination with best medical therapy, versus best medical therapy alone, in advanced Parkinson's disease. Journal of neurology. 2014;261(1): Ferreira D, Molina Y, Machado A, Westman E, Wahlund LO, Nieto A, et al. Cognitive decline is mediated by gray matter changes during middle age. Neurobiology of aging. 2014;35(5): Compta Y, Parkkinen L, Kempster P, Selikhova M, Lashley T, Holton JL, et al. The significance of α-synuclein, amyloid-β and tau pathologies in Parkinson's disease progression and related dementia. Neuro-degenerative diseases. 2014;13(2-3): Research Projects Code: 2014\FTPGB-LAGUNA Title: Estudio del perfil transcripcional en subgroups de pacientes prodrómicos con enfermedad de Parkinson Principal Investigator: Ariadna Laguna Tuset CIBERNED s collaboration: Yes Groups CIBERNED: G207; G109 Type: National Funding Agency: Fundación Tatiana Pérez de Guzmán el Bueno Funding: Duration: Code: PI2013_08-1 Title: La dinamica mitocondrial y mitofagia como dianas terapéuticas en las Enfermedades de Parkinson y Huntington Principal Investigator: Eduardo Soriano Program 2 214

221 2014 Annual Report Eduard Tolosa Sarró Group CIBERNED s collaboration: Yes Groups CIBERNED: G301; G207; G410; G109; G408 Type: National Funding Agency: ISCIII Funding: Duration: Code: Title: The Parkinson's Progression Markers Initiative (PPMI) Principal Investigator: Eduardo Tolosa Groups CIBERNED: Type: International Funding Agency: The Michael J. Fox Foundation for Parkinson's Research Funding: Duration: Code: COURAGE PD Title: Comprehensive Unbiased Risk factor Assessment for Genetics and Environment in Parkinson s Disease Principal Investigator: Thomas Gasser Groups CIBERNED: Type: International Funding Agency: JPND Funding: Duration: PhD Dissertations Author: Judith Navarro Otano Title: Estudio del Sistema Nervioso Autónomo en la enfermedad de Parkinson y otras alfasinucleinopatías Date: 15th October 2014 Supervisor: Eduardo Tolosa Sarró Author: Hugo C. Baggio Title: Brain Connectivity and cognitive impairment in Parkinson's disease Date: 14th May 2014 Supervisor: Carmen Junqué Plaja Author: Yaroslau Compta Tíitulo: Cerebrospinal fluid biomarkers in Parkinson s disease dementia. Clinico-pathological racionale and clinical correlates Date: 11th March 2014 Supervisor: María José Martí Doménech 215

222 Group Carlos Vicario Abejón Program 2 CONTACT DETAILS oup Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos CSICVicario Abejón rlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Abejón Avenida Group Doctor Carlos Arce, Vicario 37 Abejón Group icario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Madrid Abejón (Spain) Group Carlos Phone: n Group Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Fax: +34 Abejón Group 754 Carlos Vicario ejón Group Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario cvicario@cajal.csic.es Abejón Group Carlos Vicario oup Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Abejón Group Carlos Vicario Abejón LIST OF PERSONNEL Carlos Vicario Abejón Principal Investigator, CSIC Senior Scientist Eva Díaz Guerra Postdoctoral Fellow Vanesa Nieto Estévez Postdoctoral Fellow Inmaculada Crespo Galán Postdoctoral Fellow Çagla Defterali PhD student Eva Rodríguez Traver PhD student Mª José Román Lab Technician Carlos Omar Oueslati Morales Master s student Inmaculada Luque Molina Master s student 216

223 2014 Annual Report Carlos Vicario Abejón Group Summary We have obtained induced neural stem cells (inscs) derived from human induced pluripotent stem cells (ips cells or ipscs). Similar to ipscs, the inscs can be expanded in cell culture, frozen and thawed. We are using the inscs as a source of astrocytes, dopaminergic neurons (which degenerate in Parkinson s disease) as well as cortical and hippocampal neurons (affected in Alzheimer s disease) in order to investigate the cellular and molecular mechanisms altered in these neurodegenerative diseases (Vergano-Vera et al.). Our studies have shown that enhancing the levels of the transcription factor Nurr1 in neural stem cells (NSCs) isolated from the olfactory bulb (O.B.) represses proliferation and induces the formation of two populations of mature dopaminergic neurons: a) with features of the ventral mesencephalon, and b) with features of the O.B. (Vergano-Vera et al., 2014). In contrast to these results, a heterozygous deletion of the Pax6 gene reduces the incorporation and survival of dopaminergic neuron in the adult O.B. (Curto et al., 2014). We are currently studying the role of IGF-I growth factor and its receptor (IGF-IR) using a conditional knock-out specific to the Nervous System and in a mouse line in which IGF-IR levels in the dentate gyrus can be drastically reduced by the injection of retroviral particles expressing Cre recombinase. We have found that the Igf-I specific deletion produces defects on hippocampal neurogenesis in the postnataladult mice and changes the pattern of gene expression in hippocampal NSCs (Nieto-Estévez et al., submitted manuscript). We have found that functionalized graphene sheets (FGS) injected in the core of the adult mouse O.B. does not alter de novo neurogenesis or the number of neurons and astrocytes. Moreover, this nanomaterial does not provoke an evident inflammatory response supporting that FGS are a biocompatible material with neural cells in vivo (Defterali et al., manuscript under preparation). Keywords Neurogenesis, ips cells, ins cells, Parkinson s disease, Alzheimer s disease, Transcription factors, Growth factors Publications Vernal R, Diaz-Guerra E, Silva A, Sanz M, Garcia-Sanz JA. Distinct human T-lymphocyte responses triggered by Porphyromonas gingivalis capsular serotypes. Journal of clinical periodontology. 2014;41(1): Epub 2013 Nov 5. Vergano-Vera E, Diaz-Guerra E, Rodriguez-Traver E, Mendez-Gomez HR, Solis O, Pignatelli J, et al. Nurr1 blocks the mitogenic effect of FGF-2 and EGF, inducing olfactory bulb neural stem cells to adopt dopaminergic and dopaminergic-gabaergic neuronal phenotypes. Developmental neurobiology. [Epub ahead of print]. Epub 2014 Nov 28. Curto GG, Nieto-Estevez V, Hurtado-Chong A, Valero J, Gomez C, Alonso JR, et al. Pax6 is essential for the maintenance and multi-lineage differentiation of neural stem cells, and for neuronal incorporation into the adult olfactory bulb. Stem cells and development. 2014;23(23): Epub 2014 Sep

224 Research Projects Code: S2011/BMD 2336 Title: Estudio de la biología de células madre neurales para su empleo en terapia celular en enfermedades neurodegenerativas Principal Investigator: Rosario Moratalla CIBERNED s collaboration: Yes Groups CIBERNED: G305; G204; G108 Tipo: CCAA Funding Agency: Comunidad Autónoma de Madrid Funding: Duration: Code: SAF R Title: Formación de neuronas en el ser humano y ratón adultos por activación de la reprogramción celular y la neurogénesis endógena Principal Investigator: Carlos Vicario Abejón Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: Code: PI2013/01 Title: Propiedades emergentes de la relación neurona-glía que subyacen a neurodegeneración y demencia en la enfermedad de Alzheimer Principal Investigator: Ignacio Torres-Alemán CIBERNED s collaboration: Yes Groups CIBERNED: G413; G204; G409; G108; G411; G415 Type: National Funding Agency: CIBERNED Funding: Duration: Program 2 218

225 Group Miquel Vila Bover Program 2 CONTACT DETAILS Group Miquel Vila Bover Group Miquel Vila Bover Group Neurodegenerative Miquel Vila Bover Diseases Group Miquel Research Vila Group Bover Group Mique Vila Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Vall Miquel d Hebron Vila Research Bover Group Institute Miquel Vila Bove Group Miquel Vila Bover Group Miquel Vila Bover Group Mediterranean Miquel Vila Bover Building, Group First Miquel Floor, Vila Lab. Bover 102 Group Mique Passeig Vall d Hebron, Vila Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Miquel Vila Barcelona Bover Group (Spain) Miquel Vila Bove Group Miquel Vila Bover Group Miquel Vila Bover Group Miquel Vila Phone: Bover Group Miquel Vila / 3885 Bover Group Mique Fax: Vila Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Miquel Vila mvila@ir.vhebron.net Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Miquel Vila Bover Group Mique LIST OF PERSONNEL Miquel Vila Bover Principal Investigator, ICREA Research Professor Celine Perier Ramón y Cajal Researcher Jordi Bové Badell Miguel Servet Researcher Marta Martínez Vicente Miguel Servet Researcher Oriol de Fàbregues-Boixar Nebot Clinical Collaborator Iria Carballo Carbajal Postdoctoral Fellow Ainhoa Plaza Zabala Postdoctoral Fellow Ariadna Laguna Tuset Postdoctoral Fellow Ariadna Recasens Ibabe Predoctoral Fellow Sandra Franco Iborra Predoctoral Fellow Albert Torra Talavera Predoctoral Fellow Annabelle Parent Technician Thais Cuadros Arasa Technician Beatriz Rodríguez Galván Technician 219

226 Summary In 2014, we have shown that α-synuclein species contained in nigral Lewy bodies (LB) from postmortem brains of patients with Parkinson s disease (PD) have the capacity to trigger a PD-like pathological process when directly injected into the brain of mice and monkeys, including intracellular and pre-synaptic accumulations of pathological α-synuclein in different interconnected brain areas and slowly progressive axon-initiated dopaminergic nigrostriatal neurodegeneration. These pathogenic effects (i) required both human α-synuclein present in LB extracts and host expression of α-synuclein, and (ii) were not observed in animals inoculated with non-lb fractions containing soluble α-synuclein derived from the same nigral PD tissue. On the other hand, we have shown that the pro-apoptotic protein Bax, which is known to permeabilize mitochondrial membranes in PD models and is activated in PD patients, is also able to permeabilize lysosomal membranes in animal models of PD, resulting in the lysosomal-autophagic defects that occurr in this disease. Furthermore, pharmacological inhibition of Bax-induced permeabilization, in both mitochondria and lysosomes, was shown to attenuate PD-linked dopaminergic neurodegeneration in experimental PD models. These results indicate that PD-related lysosomal impairment relies on Baxinduced lysosomal membrane permeabilization and point to small molecules able to block Bax channel activity as potentially beneficial to attenuate both lysosomal defects and neurodegeneration occurring in PD. Keywords Parkinson s disease, Lewy bodies, α-synuclein, Mitochondria, Lysosomes Publications Recasens A, Dehay B. Alpha-synuclein spreading in Parkinson s disease. Frontiers in Neuroanatomy. 2014;8:159. Bove J, Martinez-Vicente M, Dehay B, Perier C, Recasens A, Bombrun A, et al. BAX channel activity mediates lysosomal disruption linked to Parkinson disease. Autophagy. 2014;10(5): Epub 2014 Mar 26. Barallobre MJ, Perier C, Bove J, Laguna A, Delabar JM, Vila M, et al. DYRK1A promotes dopaminergic neuron survival in the developing brain and in a mouse model of Parkinson s disease. Cell Death & Disease. 2014;5:e1289. Cuadros T, Trilla E, Sarro E, Vilà MR, Vilardell J, de Torres I, et al. HAVCR/KIM-1 activates the IL-6/STAT- 3 pathway in clear cell renal cell carcinoma and determines tumor progression and patient outcome. Cancer research. 2014;74(5): Epub 2014 Jan 3. Pont-Sunyer C, Hotter A, Gaig C, Seppi K, Compta Y, Katzenschlager R, et al. The Onset of Nonmotor Symptoms in Parkinson s disease (The ONSET PD Study). Movement disorders: official journal of the Movement Disorder Society. 2015;30(2): Epub 2014 Dec 1. Recasens A, Dehay B, Bove J, Carballo-Carbajal I, Dovero S, Perez-Villalba A, et al. Lewy body extracts from Parkinson disease brains trigger α-synuclein pathology and neurodegeneration in mice and monkeys. Annals of neurology. 2014;75(3): Program 2 220

227 2014 Annual Report Miquel Vila Bover Group Research Projects Code: 2014 SGR 1609 Title: Apoptosi i Neurodegeneració (GRC) Principal Investigator: Joan Comella Carnicé CIBERNED s collaboration: Yes Groups CIBERNED: G413; G109 Type: International Funding Agency: AGAUR Funding: Duration: Code: SAF R Title: Disfunción mitocondrial en la enfermedad de Parkinson: papel de la dinámica mitocondrial Principal Investigator: Celine Perier Groups CIBERNED: Type: National Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: CP11/00229 Title: Estrategias neuroprotectoras y biomarcadores en la enfermedad de Parkinson basados en la disfunción lisosomal Principal Investigator: Jordi Bove Badell Groups CIBERNED: Type: National Funding Agency: FIS-ISCIII Funding: Duration: Code: 2014\FTPGB-LAGUNA Title: Estudio del perfil transcripcional en subgroups de pacientes prodrómicos con enfermedad de Parkinson Principal Investigator: Ariadna Laguna Tuset CIBERNED s collaboration: Yes Groups CIBERNED: G207; G109 Type: National Funding Agency: Fundación Tatiana Pérez de Guzmán el Bueno Funding: Duration: Code: RTC Title: Nuevas estrategias terapéuticas para el tratamiento de sinocleunopatías Principal Investigator: Raquel Revilla (nlife Therapeutics) Groups CIBERNED: Type: National Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: MJFF TPP2014 Title: Pharmacokinetic and pharmacodynamic characterization of a novel therapy to silence selectively Principal Investigator: Raquel Revilla (nlife Therapeutics) Groups CIBERNED: Type: International Funding Agency: Michael J. Fox Foundation Funding: Duration: Code: MJFF Staff Initiated 2013 Title: Prion-like dissemination of synuclein pathology: a non-human primate study Principal Investigator: Erwan Bezard (Universite de Bordeaux, France) CIBERNED s collaboration: Yes Groups CIBERNED: G205; G109 Type: International Funding Agency: Michael J. Fox Foundation Funding: Duration: Code: Marato TV3 (120531) Title: Regenerating dopaminergic neurons in Parkinson s disease via cell-fusion-mediated reprogramming Principal Investigator: Maria Pia Cosma (CRG, Barcelona) Groups CIBERNED: Type: Private Funding Agency: Marató TV3 Funding: Duration: Code: PI13/01897 Title: Rol de la neuromelanina y alfa-sinucleína en el inicio y extensión de la Enfermedad de Parkinson: potencial diagnóstico y terapéutico en nuevos modelos pre-clínicos 221

228 Principal Investigator: Miquel Vila Bover Groups CIBERNED: Type: National Funding Agency: FIS-ISCIII Funding: Duration: Code: PI14/01529 Title: Rol patogénico de la disfunción autofágica/lisosomal en la enfermedad de Parkinson: diagnóstico y nuevas terapias pre-clínicas Principal Investigator: Marta Martinez Vicente Groups CIBERNED: Other CIBER s collaboration: Yes (CIBERBBN) Type: International Funding Agency: Fondo de Investigación Sanitaria Funding: Duration: Code: IPT Title: Suppression of alpha-synuclein expression and PDE10 as a new therapeutic strategy to Parkinson disease and improvement of cognitive dysfunctions Principal Investigator: Andres Montefeltro (nlife Therapeutics) Groups CIBERNED: Type: European Funding Agency: European Regional Development Fund, INNPACTO program Funding: Duration: Code: PI2013_08-1 Title: La dinámica mitocondrial y mitofagia como dianas terapéuticas en las Enfermedades de Parkinson y Huntington Principal Investigator: Eduardo Soriano CIBERNED s collaboration: Yes Groups CIBERNED: G301; G207; G410; G109; G408 Type: National Funding Agency: ISCIII Funding: Duration: PhD Dissertations Author: Ariadna Recasens Ibabe Title: Initiation, progression and extension of Parkinson s disease: role of α-synuclein Date: 15th June 2014 Supervisor: Miquel Vila Bover Program 2 222

229 Neuromuscular Diseases Program 3 José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo López de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Grou Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo López de Munain Group Pura Muñoz Cánoves Grou Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Group Rafael Fernández Chacó Group Isabel Illa Sendra Group Adolfo López de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebe Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendr Group Adolfo López de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilche Padilla Group José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo Lópe de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group Jos Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo López de Munain Group Pur Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo López de Munain Group Pura Muñoz Cánoves Grou Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Group Rafael Fernández Chacó Group Isabel Illa Sendra Group Adolfo López de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebe Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendr Group Adolfo López de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilche Padilla Group José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo Lópe de Munain Group Pura Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group Jos Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo López de Munain Group Pur Muñoz Cánoves Group Xavier Navarro Acebes Group Juan Jesús Vilchez Padilla Group José Berciano Blanco Group Rafael Fernández Chacón Group Isabel Illa Sendra Group Adolfo López de Munain Group Pura Muñoz Cánoves Group RESEARCH GROUPS Berciano Blanco, José Fernández Chacón, Rafael Illa Sendra, Isabel López de Munain, Adolfo Muñoz Cánoves, Pura Navarro Acebes, Xavier Vilchez Padilla, Juan Jesús

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231 2014 Annual Report Scientific Report Program 3 Neuromuscular diseases (NMDs) form a heterogeneous group of pathologies affecting the spinal cord and its tracts, nerve roots as well as motor and sensitive peripheral nerves, the neuromuscular junction and muscles. Diagnosis involves using sophisticated methods covering: neurophysiological studies, muscle or nerve biopsy with the use of immunohistochemical techniques and some others, metabolic analysis and tests, RMN studies, quantitative muscle assessment (QMA) and, in many cases, genetic studies. Just a few epidemiological studies are available in Spain, from which it can be inferred that the prevalence of chronic neurodegenerative and/or hereditary pathology would account for around 60,000 patients in our country. The members of the neuromuscular pathology program are committed to investigate into specific aspects including: Clinical characterization of neuromuscular pathologies and correlation between the clinical phenotype and their genotype (or proteomic characterization). Development of animal models based on the genotypically identified dystrophies. Research on the physiopathology of neuromuscular diseases. Development of new therapeutic strategies. The program comprises 7 clinical and basic research groups. 225

232 Groups Principal Investigator Berciano Blanco, José Fernández Chacón, Rafael Illa Sendra, Isabel * López de Munain, Adolfo Muñoz Cánoves, Pura Navarro Acebes, Xavier Vilchez Padilla, Juan Jesús * Institution Marqués de Valdecilla Hospital, Santander University of Seville, Institute of Biomedicine of Seville (IBiS), Seville Santa Creu i Sant Pau Hospital, Barcelona Biodonostia Research Institute, San Sebastián Pompeu Fabra University, Barcelona Autonomous University of Barcelona La Fe University Hospital, Valencia * These groups have voluntarily transferred to CIBERER in June 2014 Program 3 has been coordinated until June 2014 by Dr. Isabel Illa (Santa Creu i Sant Pau Hospital, Barcelona). Program 3 226

233 GROUP José Berciano Blanco Program 3 CONTACT DETAILS Group José Berciano Blanco Group José Berciano Blanco Group José Berciano Servicio Blanco Group de Neurología José Berciano Blanco José Berciano Blanco Group José Berciano Blanco Group José Hospital Berciano Universitario Blanco Group Marqués José de Valdecilla Berciano Blanco Group Berciano Blanco Group José Berciano Blanco Group José Berciano Blanco Group Universidad José Berciano de Cantabria Blanco Group Blanc Santander (Spain) Group José Berciano Blanco Group José Berciano Blanco Group jaberciano@humv.es José Berciano / combarro@unican.es Blanco Group José Berciano Blanc José Berciano Blanco Group José Berciano Blanco Group José Berciano Blanco Group José Berciano Blanco Group Blanco Group José Berciano Blanco Group José Berciano Blanco Group José Berciano Blanco Group José Berciano LIST OF PERSONNEL José Berciano Principal Investigator, Full Professor of Neurology Onofre Combarros Co-Principal Investigator, Full Professor of Neurology María Teresa Berciano Full Professor of Cell Biology Iñigo Casafont Assitant Professor Marta de la Fuente CIBERNED part-time Administrative Assistant Elena Gallardo Area Specialist Physician [ASP], Assistant Professor of Radiology Antonio García Clinical Neurophysiology ASP Isabel González-Aramburu Neurology specialist, IFIMAV postmir Fellow Andrés González-Mandly Radiology ASP Andrea González Suaréz Neurologist specialist Jon Infante ASP, Assistant Professor of Neurology Miguel A. Lafarga Full Professor of Cell Biology Jorge Mata Garrido FPI PhD student José Ignacio Mateo Neurology ASP Ana Palanca FPIMICINN PhD student Ana L. Pelayo-Negro Neurologist specialist, Lopez Albo Fellow, IDIVAL José M. Polo Associate Professor of Neurology Ana Pozueta Psychologist, FIS/IFIMAV Fellow Javier Riancho-Zarrabeitia Neurology MIR Eloy Rodríguez-Rodríguez Neurology ASP María Ruiz Soto MICINN Technician Pascual Sánchez-Juan Neurology ASP Coro Sánchez-Quintana CIBERNED Lab Technician María J. Sedano Neurology ASP María Sierra Neurology Specialist, López-Albo IFIMAV Fellow José L. Vázquez-Higuera Neurology ASP 227

234 Summary Peripheral neuropathies In 2014, a prospective study was completed in 14 patients with CMT1A versus controls, in which the clinical, neurophysiological and leg atrophy evolution illustrated by MR imaging was studied for a period of two years. This study, which was the subject of Dr. Ana L. Pelayo-Negro doctoral thesis (UC, 2014) shows that the progression of the disease is scarce, which should be taken into consideration when planning future therapeutic trials (see list of references). We have also completed a prospective clinical-neurophysiological and nerve trunks ultrasonographic study in six patients with early Guillain-Barré syndrome admitted between 2/1/2013 and 1/31/2014 (Gallardo et al., 2014). With detailed pathological correlation in a case, we demonstrated the pathogenic relevance of spinal nerves inflammation. Our work suggests that at early stages of clinical course of disease, inflammatory edema of the spinal nerves may be pathogenic because it causes an increase of the endoneural pressure compromising the transperineural blood flow, which would result in blocking the conduction with the corresponding paralysis. In the Department of Anatomy and Cell Biology at UC, Dr. Javier Riancho has set up a mouse model of ALS, hsod1 G93A analyzing the cellular mechanisms of neurodegeneration (see list of references). This research includes a therapeutic trial with bexarotene (work in progress). In collaborating with Dr. Albena Jordanova (Neurogenetics Laboratory, Antwerp University, Belgium) we are finalizing an original article in which we describe a new phenotype in the intermediate form of Charcot-Marie-Tooth disease associated with the NEFL E397K mutation. Hereditary ataxias We have continued with the European study Natural History of SCAs, which is part of the EUROSCA Project ( Also as part of EUROSCA activities, we have continued with the prospective follow-up of patients with autosomal dominant ataxia included in the RISCA Project. We have participated in the development of a predictive model of the age of onset in SCA types 1, 2, 3 and 6 (see list of references). Parkinson s disease We have shown that serum uric acid levels are not associated with the risk of developing dementia in Parkinson s disease. In two international collaborative studies, we have investigated the temporal profile of non-motor symptoms in idiopathic and LRRK2 G2019S mutation-associated Parkinson s disease (PD). By transcriptomic analysis in blood samples, we have identified 13 candidate genes in sporadic cases of parkinsonism, in asymptomatic carriers of the G2019S mutation, and controls. Within our activity as members of the EMSA-SG ( we have participated in the study that discards the pathogenic role of the COQ2 mutation in multiple systems atrophy (MSA. In collaboration with the Department of Neurology Purpam of Toulouse Hospital (INSERM), project to identify clinical and preclinical biomarkers (multimodal MRI) in LRRK2-PD is ongoing. Alzheimer s disease Together with the GIFS international consortium, we have identified two new risk loci for Alzheimer s disease (AD) on chromosomes 8 and 14, and the analysis of biological pathways involved in the disease, an increase in the expression of immune response, endocytosis, cholesterol transport and protein ubiquitination genes was observed. In collaboration with the Epistasis Project international consortium we have observed that the interaction between the gene for interleukin-10 and aromatase modifies the risk for AD in women. Program 3 228

235 2014 Annual Report José Berciano Blanco Group Our group is a founding member of the Dementia Genetics Spanish Consortium (DEGESCO) and, as a result of this collaboration, sponsored by CIBERNED, the first work of the consortium has been published, replicating in our country the association between the TREM2 gene and AD. We have also analyzed the associations between rare variants of the MAPT gene and various neurodegenerative diseases. Our group is investigating the usefulness of CSF markers and molecular imaging in patients with dementia. During 2014 we have published an article in the Journal Neurology, in collaboration with the group of Prof. Bruce Miller at UCSF, describing the clinical utility of FDG-PET and PiB-PET in patients with dementia. Prionopathies The genome-wide association (GWAs) analysis of 1,543 samples from patients with sporadic Creutzfeldt- Jakob, from 7 European countries and Australia, and 4,203 controls has been completed during The resulting manuscript entitled A genome wide association study links glutamate receptor pathway to Sporadic Creutzfeldt-Jakob disease risk is under review in the journal PLoS Genetics. We continue our collaboration with Prof. Inga Zerr, University of Goettingen, within the European project JPND DEMTEST. Cell Neurobiology of the Nucleus Studies have focused on two research lines: i) cellular basis of neuroprotection in a mouse model of ALS (SOD1- G93A transgenic mouse) and ii) compartmentalization and dynamics of foci of DNA injury/repair in sensory ganglion neurons. Regarding the first onee, we have shown that motor neurons affected in the experimental model of ALS develop neuroprotective mechanisms to extend survival and compensate for the severe translation inhibition associated with chromatolysis. Essentially, a protein synthesis machinery is reorganized de novo in the perinuclear region and a high transcriptional activity remains both in euchromatin domains (genes encoding proteins) as in the nucleolus (ribosomal genes). This response tends to enhance neuronal survival protein synthesis and ribosome biogenesis. Regarding sources of DNA injury/repair we have characterized the molecular and structural organization of the persistent foci of unrepaired DNA, which remain months after induction of DNA damage by ionizing radiation. Keywords Ataxia, SCA, Charcot-Marie-Tooth Disease, Parkinson s Disease, Alzheimer s Disease, Creutzfeldt-Jakob Disease, Nuclear Cajal Bodies, Oculopharyngeal Dystrophy Publications International Genomics of Alzheimer's Disease Consortium (IGAP), International Genomics of Alzheimer's Disease Consortium IGAP. Convergent genetic and expression data implicate immunity in Alzheimer's disease. Alzheimer's & dementia: the journal of the Alzheimer's Association. [Epub ahead of print]. Epub 2014 Dec 20. Querol L, Nogales-Gadea G, Rojas-Garcia R, Diaz-Manera J, Pardo J, Ortega-Moreno A, et al. Neurofascin IgG4 antibodies in CIDP associate with disabling tremor and poor response to IVIg. Neurology. 2014;82(10): Epub 2014 Feb 12. Medway C, Combarros O, Cortina-Borja M, Butler HT, Ibrahim-Verbaas CA, de Bruijn RF, et al. The sex-specific associations of the aromatase gene with Alzheimer's disease and its interaction with IL10 in the Epistasis Project. European journal of human genetics: EJHG. 2014;22(2): Epub 2013 Jun

236 Escott-Price V, Bellenguez C, Wang LS, Choi SH, Harold D, Jones L, et al. Gene-wide analysis detects two new susceptibility genes for Alzheimer's disease. PloS one. 2014;9(6):e Epub 2014 Jun 12. Sharma M, Wenning G, Krüger R; European Multiple-System Atrophy Study Group (EMSA-SG). Mutant COQ2 in multiple-system atrophy. The New England journal of medicine. 2014;371(1):80-1. Krasnianski A, Sanchez Juan P, Ponto C, Bartl M,Heinemann U, Varges D, et al. A proposal of new diagnostic pathway for fatal familial insomnia. Journal of neurology, neurosurgery, and psychiatry. 2014;85(6): Epub 2013 Nov 18. Riancho J, Infante J, Gonzalez-Vela C, Carril JM, Berciano J, Martinez-Rodriguez I. Acute imbalance and constitutional syndrome: the answer may lie on the front of the head. The Journal of rheumatology. 2014;41(1): Pelayo-Negro AL, Carr AS, Laura M, Skorupinska M, Reilly MM. An observational study of asymmetry in CMT1A. Journal of neurology, neurosurgery, and psychiatry. [Epub ahead of print]. Epub 2014 Oct 13. Gazulla J, Mayayo-Sinues E, Benavente I, Modrego PJ, Berciano J. Ataxia of Charlevoix-Saguenay: MR and Clinical Results in Lower-Limb Musculature. The Canadian journal of neurological sciences. Le journal canadien des sciences neurologiques. 2014;41(1): Berciano J. Commentary. Movement disorders: official journal of the Movement Disorder Society. 2014;29(1):38-9. Epub 2013 Dec 16. Riancho J, Ruiz-Soto M, Villagra NT, Berciano J, Berciano MT, Lafarga M. Compensatory Motor Neuron Response to Chromatolysis in the Murine hsod1(g93a) Model of Amyotrophic Lateral Sclerosis. Frontiers in cellular neuroscience. 2014;8:346. Epub 2014 Oct 22. Garcia A, Pelayo-Negro AL, Alvarez-Paradelo S, Antolin FM, Berciano J. Electromyographic tendon reflex recording: An accurate and comfortable method for diagnosis of Charcot-Marie-Tooth disease type 1A. Muscle & nerve. [Epub ahead of print]. Epub 2014 Oct 31. Pelayo-Negro AL, Gallardo E, Garcia A, Sanchez-Juan P, Infante J, Berciano J. Evolution of Charcot- Marie-Tooth disease type 1A duplication: a 2-year clinico-electrophysiological and lower-limb muscle MRI longitudinal study. Journal of neurology. 2014;261(4): Epub 2014 Jan 22. Infante J, Prieto C, Sierra M, Sanchez-Juan P, Gonzalez-Aramburu I, Sanchez-Quintana C, et al. Identification of candidate genes for Parkinson's disease through blood transcriptome analysis in LRRK2-G2019S carriers, idiopathic cases, and controls. Neurobiology of aging. 2015;36(2): Epub 2014 Nov 5. Tezenas du Montcel S, Durr A, Bauer P, Figueroa KP, Ichikawa Y, Brussino A, et al. Modulation of the age at onset in spinocerebellar ataxia by CAG tracts in various genes. Brain: a journal of neurology. 2014;137(Pt 9): Epub 2014 Jun 26. Berciano J, Gallardo E, Sedano MJ, Orizaola P, Sanchez-Juan P, Gonzalez-Suarez A, et al. Nerve ultrasonography in early Guillain-Barre syndrome: a need for large prospective studies. Journal of the Peripheral Nervous System. 2014;19:344. Epub Gaig C, Vilas D, Infante J, Sierra M, Garcia-Gorostiaga I,Buongiorno M, et al. Nonmotor symptoms in LRRK2 G2019S associated Parkinson's disease. PloS one. 2014;9(10):e Epub 2014 Oct 17. Sanchez-Juan P, Ghosh PM, Hagen J, Gesierich B, Henry M, Grinberg LT, et al. Practical utility of amyloid and FDG-PET in an academic dementia center. Neurology. 2014;82(3): Epub 2013 Dec 18. Tezenas du Montcel S, Durr A, Rakowicz M, Nanetti L, Charles P, Sulek A, et al. Prediction of the age at onset in spinocerebellar ataxia type 1, 2, 3 and 6. Journal of medical genetics. 2014;51(7): Epub 2014 Apr 29. Palanca A, Casafont I, Berciano MT, Lafarga M. Proteasome inhibition induces DNA damage and reorganizes nuclear architecture and protein synthesis machinery in sensory ganglion neurons. Cellular and molecular life sciences: CMLS. 2014;71(10): Epub 2013 Sep 24. Palanca A, Casafont I, Berciano MT, Lafarga M. Reactive nucleolar and Cajal body responses to proteasome inhibition in sensory ganglion neurons. Biochimica et biophysica acta. 2014;1842(6): Epub 2013 Nov 19. Mannil M, Solari A, Leha A, Pelayo-Negro AL, Berciano J, Schlotter-Weigel B, et al. Selected items from the Charcot-Marie-Tooth (CMT) Neuropathy Score and secondary clinical outcome measures serve as sensitive clinical markers of disease severity in CMT1A patients. Neuromuscular disorders: Program 3 230

237 2014 Annual Report José Berciano Blanco Group NMD. 2014;24(11): Epub 2014 Jun 19. Gonzalez-Aramburu I, Sanchez-Juan P, Sierra M, Fernandez-Juan E, Sanchez-Quintana C, Berciano J, et al. Serum uric acid and risk of dementia in Parkinson's disease. Parkinsonism & related disorders. 2014;20(6): Epub 2014 Mar 5. Gallardo E, Sedano MJ, Orizaola P, Sanchez-Juan P, Gonzalez-Suarez A, Garcia A, et al. Spinal nerve involvement in early Guillain-Barre syndrome: A clinico-electrophysiological, ultrasonographic and pathological study. Clinical neurophysiology: official journal of the International Federation of Clinical Neurophysiology. [Epub ahead of print]. Epub 2014 Aug 21. Pont-Sunyer C, Hotter A, Gaig C, Seppi K, Compta Y, Katzenschlager R, et al. The Onset of Nonmotor Symptoms in Parkinson's disease (The ONSET PD Study). Movement disorders: official journal of the Movement Disorder Society. 2015;30(2): Epub 2014 Dec 1. Tapia O, Lafarga V, Bengoechea R, Palanca A, Lafarga M, Berciano MT. The SMN Tudor SIM-like domain is key to SmD1 and coilin interactions and to Cajal body biogenesis. Journal of cell science. 2014;127(Pt 5): Epub 2014 Jan 10. Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodriguez-Rodriguez E, Lopez de Munain A, et al. Assessing the role of the TREM2 p.r47h variant as a risk factor for Alzheimer's disease and frontotemporal dementia. Neurobiology of aging. 2014;35(2):444.e1-4. Riancho-Zarrabeitia L, Delgado-Alvarado M, Riancho J, Oterino A, Sedano MJ, Rueda-Gotor J, et al. Anti-TNF-α therapy in the management of severe neurosarcoidosis: a report of five cases from a single centre and literature review. Clinical and experimental rheumatology. 2014;32(2): Delgado-Alvarado M, Gomez-Roman J, Sanchez-Salmon E, Rodriguez-Rodriguez E, Polo JM, Garcia- Castano A, et al. Nonanaplastic pleomorphic xanthoastrocytoma with meningeal dissemination presenting with bilateral visual loss. Journal of neuroimaging: official journal of the American Society of Neuroimaging. 2014;24(5): Research Projects Code: PI11/03028 Title: DEMTEST: Biomarker based diagnosis of rapid progressive dementias Principal Investigator: Pascual Sánchez CIBERNED s collaboration: Yes Groups CIBERNED: G601; G114; G503; G609 Type: Others Funding Agency: Instituto de Salud Carlos III - Comunidad Económica Europea Funding: Duration: Code: IFIMAV/LOPEZ ALBO 2013 Title: Amiotrofia de extremidades inferiores en la enfermedad de Charcot-Marie-Tooth tipo 1A: estudio longitudinal clínico, neurofisiológico y de resonancia magnética Principal Investigator: Ana Lara Pelayo Groups CIBERNED: Type: International Funding Agency: IDIVAL Funding: N.D. Duration: 2014 Code: PI13/01008 Title: Biomarcadores de enfermedad de Alzheimer como factores pronóstico en hidrocefalia a presión normal idiomática Principal Investigator: Eloy Rodríguez Groups CIBERNED: Type: International Funding Agency: ISCIII Funding: N.D. Duration: 2014 Code: FI11/00259 Title: Enfermedad de Parkinson causada por la mutación G2019S de LRRK2 en Cantabria: aspectos genéticos y estudio de biomarcadores en portadores asintomáticos de la mutación Principal Investigator: María Sierra Peña Groups CIBERNED: 231

238 Type: National Funding Agency: ISCIII Funding: Duration: Code: WLA 04/11 Title: Estudio de biomarcadores en la fase promotora en la enfermedad de Parkinson genética y su aplicación al diagnóstico de la enfermedad de Parkinson idiopática Principal Investigator: María Sierra Peña Groups CIBERNED: Type: National Funding Agency: Fundación Marqués de Valdecilla IFIMAV Funding: Duration: Code: IFIMAV/LOPEZ ALBO Title: Estudio de biomarcadores en la fase promotora en la enfermedad de Parkinson genética y su aplicación al diagnóstico de la enfermedad de Parkinson idiopática Principal Investigator: María Sierra Peña Groups CIBERNED: Type: International Funding Agency: IDIVAL Funding: N.D. Duration: 2014 Code: FI12/00229 Title: Estudio de la influencia de los factores genéticos reguladores de los niveles séricos de Progranulina sobre la penetrancia de la mutación G2019S de LRRK2 y sobre el riesgo de demencia en la enfermedad de Parkinson Principal Investigator: Isabel González Aramburu Groups CIBERNED: Type: International Funding Agency: ISCIII Funding: N.D. Duration: 2014 Code: PI12/02288 Title: Estudio multimodal de biomarcadores de enfermedad de Alzheimer en deterioro cognitivo postoperatorio Principal Investigator: Pascual Sánchez-Juan Groups CIBERNED: Type: National Funding Agency: FIS Funding: Duration: Code: Ayuda del Comité ad-hoc de Neurólogos Jóvenes de la SEN Title: Identificación mediante resonancia multimodal de biomarcadores de la fase premotora de la enfermedad de Parkinson asociada a la mutación G2019S de LRRK2 (proyecto de colaboracion con Hospital Purpan de Toulouse y con el INSERM, Francia) Principal Investigator: María Sierra Peña Groups CIBERNED: Type: International Funding Agency: Sociedad Española de Neurología Funding: N.D. Duration: 2014 Code: JPND Title: Proyecto DEMTEST Principal Investigator: Pascual Sánchez-Juan Groups CIBERNED: Type: National Funding Agency: FIS Funding: Duration: Code: RD09/0076/00076 Title: RETIC Biobancos, nodo Hospital "Universitario Marqués de Valdecilla" Principal Investigator: Pascual Sánchez Juan Groups CIBERNED: Type: National Funding Agency: ISCIII Funding: Duration: Code: PI11/00228 Title: Selección de genes candidato para la enfermedad de Parkinson idiopática a través del análisis diferencial del transcriptoma en pacientes y portadores asintomáticos de mutaciones en LRRK2 Principal Investigator: Jon Infante Ceberio Groups CIBERNED: Type: National Funding Agency: ISCIII Funding: Duration: Code: BFU Title: Señalización y reparación del daño en el DNA en neuronas. Organización estructural, molecular, espacial y Program 3 232

239 2014 Annual Report José Berciano Blanco Group temporal de los focos de lesión/reparación del DNA Principal Investigator: Miguel Angel Lafarga Groups CIBERNED: Type: National Funding Agency: Dirección General de Investigación Funding: Duration: PhD Dissertations Author: Ana Palanca Cuñado Title: Respuesta de las neuronas sensitivas al estrés genotóxico inducido por radiaciones ionizantes y por la inhibición del proteasoma Date: 13th June 2014 Supervisor: María Teresa Berciano Blanco 233

240 GROUP Rafael Fernández Chacón Program 3 CONTACT DETAILS oup Rafael Fernández Chacón Group Rafael Fernández Chacón Instituto Group de Biomedicina Rafael Fernández Sevilla Chacón (IBiS) Group Rafael rnández Chacón Group Rafael Fernández Hospital Chacón Universitario Group Rafael Virgen Fernández del Rocío/CSIC/Universidad Chacón Group Rafael de Sevilla Fernández Chacón oup Rafael Fernández Chacón Departamento Group Rafael de Fernández Fisiología Médica Chacón y Group Biofísica de Rafael la Universidad Fernández de Chacón Sevilla Group Rafael CIBERNED rnández Chacón Group Rafael Fernández Chacón Group Rafael Fernández Chacón Avenida Group Manuel Siurot, Rafael s/n Fernández Chacón oup Rafael Fernández Chacón Group Rafael Fernández Chacón Group Rafael Fernández Sevilla Chacón (Spain) Group Rafael Phone: (office) / (laboratorio) rnández Chacón Group Rafael Fernández Chacón Group Rafael Fernández Chacón Fax: Group Rafael Fernández Chacón oup Rafael Fernández Chacón Group Rafael Fernández Chacón Group Rafael Fernández rfchacon@us.es Chacón Group Rafael rnández Chacón Group Rafael Fernández Chacón Group Rafael Fernández Chacón Group Rafael Fernández Chacón LIST OF PERSONNEL LIST OF PERSONNEL Rafael Fernández Chacón Principal Investigator, Associate Professor Ana Cristina Calvo Laínez Postdoctoral researcher Fabiola Mavillard Saborido Postdoctoral researcher Carmen Paradas Postdoctoral researcher Gloria Cantero Nieto Postdoctoral researcher José Luis Rozas Espadas Postdoctoral researcher Josif Mircheski Postdoctoral researcher Pablo Luis García-Junco Clemente Postdoctoral researcher José Antonio Martínez López Predoctoral researcher Leonardo Gómez Sánchez Predoctoral researcher Macarena Cabrera Serrano Predoctoral researcher Angela Lavado Roldán Predoctoral researcher Alejandro Arroyo Saborido Technician María del Carmen Rivero Mena Technician Jose Luis Muñoz Bravo Researcher 234

241 2014 Annual Report Rafael Fernández Chacón Group Summary Our group studies the molecular basis of synaptic function and maintenance of nerve terminals. In 2014, using functional imaging techniques (synaptophluorin) and electron microscopy in transgenic mice we have concluded a study focused on important details of the dynamic and pharmacology of synaptic vesicle endocytosis in motor nerve terminals (article under review). On the other hand, Dr. Jose Luis Nieto-González, in a project led by Prof. Gómez-Scholl (IBiS, Sevilla), has studied the function of central synapses in transgenic mice expressing a mutated form of beta-neurexin 1, a synaptic protein associated to autism in humans. These mice develop autistic symptoms due to beta-neurexin 1 dysfunction. Interestingly, those symptoms are reversed upon switching off the expression of such a mutant version of beta-neurexin 1 even in mice relatively old (Rabaneda et al., 2014). A major goal in our lab is to elucidate the role of Cysteine String Protein-alpha (CSP-alpha), a molecular co-chaperone located at the synaptic vesicles that prevents synaptic degeneration. There are two mutations in the human gene encoding CSP-alpha (DNAJC5) causing autosomal-dominant adult-onset neuronal ceroid lipofuscinosis (NCL4), a devastating neurodegenerative disease in young adults. En 2014, we have initiated the analysis of novel transgenic mice lines that express those mutations in different areas of the central nervous system. In addition, we have described extrasynaptic functions of CSP-alpha such as a very interesting role in postnatal neurogenesis. Furthermore, within out group, Dr. Carmen Paradas, leads a research line focused on the genetic basis of muscular dystrophies in humans. An important progress achieved in 2014 has been the design and generation of genetically modifed mice bearing a mutation in a new gene causing familial limb-girdle muscular dystrophy. That mutation causes a deficit in Notch receptor activation that could intefere with the regenerative function of satellite cells, the muscle stem cells. Keywords Synapse, Synaptic vesicle cycle, Neurogenesis, Neuronal ceroid lipofuscinosis, Muscular dystrophies, Human genetics, Genetically modified mice Publications Cabrera-Serrano M, Fabian VA, Boutilier J, Wise C, Faiz F, Lamont PJ, et al. Adult onset distal and proximal myopathy with complete ophthalmoplegia associated with a novel de novo p.(leu1877pro) mutation in MYH2. Clinical genetics. [Epub ahead of print]. Epub 2014 Dec 22. Ohkawara B, Cabrera-Serrano M, Nakata T, Milone M, Asai N, Ito K, et al. LRP4 third β-propeller domain mutations cause novel congenital myasthenia by compromising agrin-mediated MuSK signaling in a position-specific manner. Human molecular genetics. 2014;23(7): Epub 2013 Nov 13. Rabaneda LG, Robles-Lanuza E, Nieto-Gonzalez JL, Scholl FG. Neurexin dysfunction in adult neurons results in autistic-like behavior in mice. Cell reports. 2014;8(2): Epub 2014 Jul 10. Cabrera-Serrano M, Junckerstorff RC, Atkinson V, Sivadorai P, Allcock RJ, Lamont P, et al. Novel CHKB mutation expands the megaconial muscular dystrophy phenotype. Muscle & nerve. 2015;51(1): Epub 2014 Nov 22. Paradas C, Akman HO, Ionete C, Lau H, Riskind PN, Jones DE, et al. Branching enzyme deficiency: expanding the clinical spectrum. JAMA neurology. 2014;71(1):

242 Research Projects Code: BFU P Title: Mecanismos moleculares y celulares de la respuesta cerebral y neuronal a la degeneración presináptica Principal Investigator: Rafael Fernández-Chacón Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: Code: P12-CTS-2232 Title: Análisis genético y funcional de co-chaperones sinápticos en el mantenimiento y la degeneración sináptica Principal Investigator: Rafael Fernández-Chacón Groups CIBERNED: Type: National Funding Agency: Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía Funding: Duration: Program 3 236

243 GROUP Isabel Illa Sendra Program 3 CONTACT DETAILS Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Hospital Sendra de Group la Santa Isabel Creu Illa i Sant Sendra PauGroup Isabel Ill Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Neurología Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel 4º Illa Piso. Sendra Edificio Group Nuevo Isabel Illa Sendr Avenida Pare Claret, 167 Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Barcelona Illa Sendra (Spain) Group Isabel Ill Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Phone: Group Isabel Illa Sendra Group Fax: Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa iilla@santpau.cat Sendra Group Isabel Illa Sendr Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Illa Sendra Group Isabel Ill LIST OF PERSONNEL Isabel Illa Sendra Principal Investigator, Full Professor Eduardo Gallardo Vigo Researcher Institut de Recerca Sant Pau Ricardo Rojas García Researcher Neurology Jordi Díaz Manera Researcher Neurology Gisela Nogales Gadea Postdoctoral researcher Sara Borrel FIS Xavier Suárez Calvet Predoctoral researcher FIS Luis Querol Gutiérrez Post MIR FIS, Researcher Río Hortega Aida Alejaldre Monforte Predoctoral researcher CIBERNED Alba Ramos Fransi Predoctoral researcher Josefa Araque Palacios Technician CIBERNED Esther Ortiz Losada Technician Institut de Recerca Sant Pau Miquel Navas Madroñal Technician FIS Sonia Segovia Simón Nurse. Project manager CIBERNED Noemi de Luna Researcher CIBERNED 237

244 Summary Immune-mediated neuromuscular diseases Immunopathogenesis studies with characterization of autoantibodies to new targets and their clinical correlation. Its use as biomarkers. Role of autoantibodies in the immunopathogenesis of CIDP and MG. Role of innate immunity in Guillain Barre Syndrome, Myasthenia, Inflammatory myopathies. Analysis of the effect of new immuno-modulating therapies on functional aspects of immune system cells (response to ligands, production of antibodies, etc.) and clinical response. Muscular dystrophy, dysferlinopathy and distal myopathies Clinical characterization, new diagnostic tests for dysferlinopathy and distal myopathies, and study of molecular pathways involved in differentiation and molecular and functional interactions of dysferlin. Amyotrophic lateral sclerosis (ALS). Epidemiology. Genetic studies. New therapies in neuromuscular diseases Therapy with new biological agents in subgroups of immune mediated NMD. Allogenic transplantation in MNGIE. Bone marrow transplantation in dysferlinopathy (animal model). Advanced therapy with mesoangioblasts. Evaluate the effects of treatment with vitamin D in carriers of a DYSF gene mutation. Challenges Advance knowledge of the immunological mechanisms involved in the pathogenesis of immunemediated neuromuscular diseases. Role of Innate immunity. Search for new antigens using new tissue sources. Development of diagnostic tests using the new antigens. Finding new and more specific immune therapies. Analysis of the natural history of dysferlinopathies. Implement new diagnostic methods for limb girdle muscular dystrophies (LGMDs) and distal myopathies. Discovery of new genes causing limb girdle muscular dystrophy. Gain knowledge on the pathogenetic mechanisms involved in muscular dystrophy resulting from a dysferlin deficiency. Set up of a clean room to manipulate cells for treatment in humans. Serum and skeletal muscle mirna profiling of limb girdle muscular dystrophies. Description of skeletal muscle MRI patterns specific for different LGMDs. Program 3 238

245 2014 Annual Report Isabel Illa Sendra Group Keywords Immune neuromuscular diseases (CIDP, MMN, MG, IM), Pathogenesis of Muscular dystrophies, Biological and cell therapies for neuromuscular diseases, ALS Publications Querol L, Nogales-Gadea G, Rojas-Garcia R, Diaz-Manera J, Pardo J, Ortega-Moreno A, et al. Neurofascin IgG4 antibodies in CIDP associate with disabling tremor and poor response to IVIg. Neurology. 2014;82(10): Epub 2014 Feb 12. Suarez-Calvet X, Gallardo E, Nogales-Gadea G, Querol L, Navas M, Diaz-Manera J, et al. Altered RIG-I/ DDX58-mediated innate immunity in dermatomyositis. The Journal of pathology. 2014;233(3): Epub 2014 Apr 29. Nogales-Gadea G, Ramos-Fransi A, Suarez-Calvet X, Navas M, Rojas-Garcia R, Mosquera JL, et al. Analysis of serum mirna profiles of myasthenia gravis patients. PloS one. 2014;9(3):e Epub 2014 Mar 17. Gallardo E, Martinez-Hernandez E, Titulaer MJ, Huijbers MG, Martinez MA, Ramos A, et al. Cortactin autoantibodies in myasthenia gravis. Autoimmunity reviews. 2014; 13(10): Epub 2014 Sep 3. Diaz-Manera J, Alejaldre A, Llauger J, Mirabet S, Rojas-Garcia R, Ramos-Fransi A, et al. Cranial, axial and proximal myopathy and hypertrophic cardiomyopathy caused by a mutation in the globular head region of the MYH7 gene. European journal of neurology: the official journal of the European Federation of Neurological Societies. 2014;21(6):e51-2. Gallardo E, Ankala A, Nunez-Alvarez Y, Hegde M, Diaz-Manera J, Luna ND, et al. Genetic and epigenetic determinants of low dysferlin expression in monocytes. Human mutation. 2014;35(8): Epub 2014 Jun 24. Aragonès JM, Roura-Poch P, Hernandez-Ocampo EM, Alonso F, Pont-Lluelles M, Xandri I, et al. Myasthenia gravis: a disease of the very old. Journal of the American Geriatrics Society. 2014;62(1): Aragonès JM, Altimiras J, Roura P, Alonso F, Bufill E, Munmany A, et al. Prevalence of myasthenia gravis in the Catalan county of Osona. Neurologia (Barcelona, Spain). [Epub ahead of print]. Epub 2014 Nov 11. Diaz-Manera J, Querol L, Alejaldre A, Rojas-Garcia R, Ramos-Fransi A, Gallardo E, et al. Severe exacerbation of Andersen-Tawil syndrome secondary to thyrotoxicosis. Journal of human genetics. 2014;59(8): Epub 2014 May 22. Labasque M, Hivert B, Nogales-Gadea G, Querol L, Illa I, Faivre-Sarrailh C. Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. The Journal of biological chemistry. 2014;289(11): Epub 2014 Feb 4. Dols-Icardo O, Garcia-Redondo A, Rojas-Garcia R, Sanchez-Valle R, Noguera A, Gomez-Tortosa E, et al. Characterization of the repeat expansion size in C9orf72 in amyotrophic lateral sclerosis and frontotemporal dementia. Human molecular genetics. 2014;23(3): Evilä A, Vihola A, Sarparanta J, Raheem O, Palmio J, Sandell S, et al. Atypical phenotypes in titinopathies explained by second titin mutations. Annals of neurology. 2014;75(2):

246 Research Projects Code: FIS PI 13/00772 Title: Base de datos española de enfermedades neuromusculares Principal Investigator: Ricardo Rojas García Groups CIBERNED: Type: International Funding Agency: Instituto de Salud Carlos III Funding: N.D. Duration: 2014 Code: PI 12/02991 Title: Búsqueda de biomarcadores de distrofias musculares con debilidad de cinturas Principal Investigator: Eduardo Gallardo Groups CIBERNED: Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: Dysferlinopathy 2012 Title: Clinical Outcome Study for Dysferlinopathy Principal Investigator: Isabel Illa Sendra Groups CIBERNED: Type: International Funding Agency: Jain Foundation Funding: Duration: Code: FIS PI/13/00937 Title: Nuevas reatividades antigénicas y estudios de inmunidad innata en enfermedades neuromusculares Principal Investigator: Isabel Illa Sendra Groups CIBERNED: Type: International Funding Agency: Instituto de Salud Carlos III Funding: N.D. Duration: 2014 Code: 2009 SGR1004 Title: Research on neuromuscular diseases. Grup de recerca consolidat de la Generalitat Principal Investigator: Isabel Illa Sendra Groups CIBERNED: Type: National Funding Agency: Generalitat de Catalunya Funding: Duration: Program 3 240

247 GROUP Adolfo López de Munain Program 3 Group Adolfo López de Munain Group Adolfo LIST OF PERSONNEL LIST OF PERSONNEL López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo CONTACT DETAILS López de Munain Group Adolfo López de Munain Instituto de Investigación Biodonostia Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López d Paseo Dr. Beguiristain, s/n Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo San Sebastián López de (Spain) Munain Group Adolf López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Leyre Curto Adolfo ArzacLópez de Munai Phone: Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo Lópe leyre.curtoarzac@osakidetza.net de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Group Adolfo López de Munain Principal Investigator María Cruz Rodríguez Oroz Researcher Ana Aiastui Pujana Researcher Garazi Aldanondo Aristizabal Researcher Miren Karmele Arnaiz Pérez Researcher Myriam Barandiarán Amillano Researcher Alberto Bergareche Yarza Researcher Cristina Caballero Martínez Researcher M. Pilar Camaño González Researcher Manuel Delgado Alvarado Researcher Ainara Estanga Alustiza Researcher Juan Bautista Espinal Valencia Researcher Roberto Fernandez Torrón Researcher Belén Gago Calderón Researcher Federico García Bragado Researcher Patricia Garcia Parra Researcher Francisco Javier Gil Bea Researcher María Goicoechea Bianchi Researcher Ana Gorostidi Pagola Researcher Begoña Indacoechea Juanbeltz Researcher Jaione Lacalle Prieto Researcher José Félix Martí Masso Researcher Pablo Martinez-Lage Álvarez Researcher Alba Mateos Mateos Ayerdi Researcher Aroa Méndez Zabalo Researcher Elisabeth Mondragón Rezola Researcher Fermín Moreno Izco Researcher Neia Naldaiz Gastezi Researcher Nahikari Pastoriza Polo Researcher Juanjo Poza Aldea Researcher Patricia de la Riva Juez Researcher Javier Ruiz Martínez Researcher Amets Sáenz Peña Researcher Ernesto Sanz Arigita Researcher Andone Sistiaga Berrondo Researcher Ivan Toral Ojeda Researcher Miguel Ángel Urtasun Ocariz Researcher Ainara Vallejo Illarramendi Researcher Miren Zulaica Ijurco Researcher Olaia Zuriarráin Bergara Researcher 241

248 Summary The main research lines are: Impulse control disorders (ICD) and dyskinesia: Physiopathology, risk factors and therapeutic approaches in these complications derived from chronic dopaminergic treatment in Parkinson s disease. We focus on clinical and experimental research in animal models with several approaches comprising behavioural, neuropsychological, molecular, genetic and image studies (PET using different radiotracers and multimodal MRI). A multicentre clinical trial on ICD coordinated by this group is also under development. Dementia and mild cognitive impairment (MCI) in PD. This line is focused on the identification of biomarkers of MCI that might be used in the early diagnosis of dementia in PD as well as in the implementation of early therapeutic interventions. In this regard, cognitive stimulation is also one of the lines of study at this moment. The approach to this line of research includes multimodal MRI, PET, electrophysiological and molecular studies in CSF and plasma as well as proteomic and epigenetic studies. Genetic and premotor biomarkers of PD. This line is focused on the study of the phenotype of patients with the LRRK2 R1441G mutation (frequent in the Basque country) and of asymptomatic carriers of this mutation aiming to know pre-motor biomarkers of PD including clinical, molecular, (CSF and plasma) and image (DATSCAN and fmri) studies as well as their evolution. Familiar esencial tremor. This line comprises clinical, neurophysiológical, genetic (exoming and genetic linkage analysis), and image (MRI) studies. The objective is to study genetic, molecular functional abnormalities in families with different clinical phenotypes. Inflammation in PD. It comprises studies in animal models of parkinsonism and in ips derived from patients with PD. The aim is to study the relationship between inflammation and dopaminergic death in animal models using in vivo PET and postmortem studies. On the other hand, the study of ips derived from genetic forms (patients and asymptomatic carriers of the LRRK2 R1441G mutation) and sporadic cases of PD will allow to deepen in the relationship between glial cells, neuroinflammation and neuronal death in both sporadic and genetic forms of PD. Neuromuscular Diseases The neuromuscular group is composed by neurologists, neuropsychologists and biologists who develop a comprehensive approach to neuromuscular pathology (including motor neuron disease, spinal atrophy, polyneuropathies, endplate diseases and muscular diseases) both in basic and clinical care. The group is also interested in the role of muscle metabolic control and its influence on the development of central phenomena associated with aging (aging brain and proteinopathies). Current research lines of the group are focused on the pathophysiology of the main forms of muscular dystrophy (myotonic dystrophy, limb girdle dystrophy type 2A, facioscapulohumeral dystrophy and Duchenne dystrophy), and on abnormal muscle homeostasis in aging and brain neurodegeneration. Myotonic dystrophy: study of the pathways that determine the risk of oncogenicity and brain disorders. Duchenne dystrophy: study of alterations in calcium homeostasis and its rescue with compounds targeting ryanodine receptor stability. Facioscapulohumeral dystrophy: study of genomic and epigenetic mechanisms that regulate the phenotypic expression in scapuloperoneales syndromes. Limb Girdle Dystrophy type 2 A: study of the influence of calpain 3 in proliferation, differentiation and replacement of muscle satellite cells. Program 3 242

249 2014 Annual Report Adolfo López de Munain Group Keywords Parkinson s disease, LRRK2, Dyskinesias, Impulse control disorder, Dementia, Cognitive impairment, Neuroinflammation, Muscular dystrophies, Frontotemporal dementia, Calpain, Progranulin, Biomarkers, Neurogenetics Publications Yakhine-Diop SM, Bravo-San Pedro JM, Gomez-Sanchez R, Pizarro-Estrella E, Rodriguez-Arribas M, Climent V, et al. G2019S LRRK2 mutant fibroblasts from Parkinson's disease patients show increased sensitivity to neurotoxin 1-methyl-4-phenylpyridinium dependent of autophagy. Toxicology. 2014;324:1-9. Epub 2014 Jul 10. Alcalay RN, Aasly J, Berg D, Bressman S, Brice A, Brockmann K, et al. Michael J. Fox Foundation LRRK2 Consortium: geographical differences in returning genetic research data to study participants. Genetics in medicine: official journal of the American College of Medical Genetics. 2014;16(8): Thelen M, Razquin C, Hernandez I, Gorostidi A, Sanchez-Valle R, Ortega-Cubero S, et al. Investigation of the role of rare TREM2 variants in frontotemporal dementia subtypes. Neurobiology of aging. 2014;35(11):2657.e13-9. Epub 2014 Jun 20. Tolosa E, Litvan I, Höglinger GU, Burn D, Lees A, Andres MV, et al. A phase 2 trial of the GSK-3 inhibitor tideglusib in progressive supranuclear palsy. Movement disorders: official journal of the Movement Disorder Society. 2014;29(4): Epub 2014 Feb 14. Manterola L, Guruceaga E, Gallego Perez-Larraya J, Gonzalez-Huarriz M, Jauregui P, Tejada S, et al. A small noncoding RNA signature found in exosomes of GBM patient serum as a diagnostic tool. Neuro-oncology. 2014;16(4): Epub 2014 Jan 16. Arruti M, Aldazabal M, Blanco A, Pineiro L, Zapico MS, Gonzalez F, et al. Acute herpes simplex virus type 1 retinal necrosis three years after herpes simplex encephalitis. Revista de neurologia. 2014;58(1):45-6. Mateos-Aierdi A, Aiastui A, Goicoechea M, Lopez de Munain A. Advances in gene therapies for limbgirdle muscular dystrophies. Advances in Regenerative Biology. 2014;1: Abete I, Arriola L, Etxezarreta N, Mozo I, Moreno-Iribas C, Amiano P, et al. Association between different obesity measures and the risk of stroke in the EPIC Spanish cohort. European journal of nutrition. [Epub ahead of print]. Epub 2014 Jun 6. Abete I, Romaguera D, Vieira AR, Lopez de Munain A, Norat T. Association between total, processed, red and white meat consumption and all-cause, CVD and IHD mortality: a meta-analysis of cohort studies. The British journal of nutrition. 2014;112(5): Epub 2014 Jun 16. Munoz-Culla M, Irizar H, Castillo-Trivino T, Saenz-Cuesta M, Sepulveda L, Lopetegi I, et al. Blood mirna expression pattern is a possible risk marker for natalizumab-associated progressive multifocal leukoencephalopathy in multiple sclerosis patients. Multiple sclerosis (Houndmills, Basingstoke, England). 2014;20(14): Epub 2014 May 22. Saenz-Cuesta M, Irizar H, Castillo-Trivino T, Munoz-Culla M, Osorio-Querejeta I, Prada A, et al. Circulating microparticles reflect treatment effects and clinical status in multiple sclerosis. Biomarkers in medicine. 2014;8(5): Kalia LV, Lang AE, Hazrati LN, Fujioka S, Wszolek ZK, Dickson DW, et al. Clinical Correlations With Lewy Body Pathology in LRRK2-Related Parkinson Disease. JAMA neurology. 2015;72(1): Epub Estanga A, Rodriguez-Oroz MC, Ruiz-Martinez J, Barandiaran M, Gorostidi A, Bergareche A, et al. Cognitive dysfunction in Parkinson's disease related to the R1441G mutation in LRRK2. Parkinsonism & related disorders. 2014;20(10): Epub 2014 Jul 30. Sistiaga A, Castillo-Trivino T, Aliri J, Gaztanaga M, Acha J, Arruti M, et al. Cognitive performance and 243

250 quality of life in multiple sclerosis in Gipuzkoa. Revista de neurologia. 2014;58(8): Suarez-Boomgaard D, Gago B, Valderrama-Carvajal A, Roales-Bujan R, Craenenbroeck KV, Duchou J, et al. Dopamine D4 receptor counteracts morphine-induced changes in µ opioid receptor signaling in the striosomes of the rat caudate putamen. International journal of molecular sciences. 2014;15(1): Epub 2014 Jan 21. Vallejo-Illarramendi A, Toral-Ojeda I, Aldanondo G, Lopez de Munain A. Dysregulation of calcium homeostasis in muscular dystrophies. Expert reviews in molecular medicine. 2014;16:e16. Epub 2014 Oct 8. Jaka O, Azpitarte M, Paisan-Ruiz C, Zulaika M, Casas-Fraile L, Sanz R, et al. Entire CAPN3 gene deletion in a patient with limb-girdle muscular dystrophy type 2A. Muscle & nerve. 2014;50(3): Epub 2014 Aug 5. Gomez-Uriz AM, Goyenechea E, Campion J, de Arce A, Martinez MT, Puchau B, et al. Epigenetic patterns of two gene promoters (TNF-α and PON) in stroke considering obesity condition and dietary intake. Journal of physiology and biochemistry. 2014;70(2): Epub 2014 Feb 7. Alvarez-Castelao B, Gorostidi A, Ruiz-Martinez J, Lopez de Munain A, Castano JG. Epitope Mapping of Antibodies to Alpha-Synuclein in LRRK2 Mutation Carriers, Idiopathic Parkinson Disease Patients, and Healthy Controls. Frontiers in aging neuroscience. 2014;6:169. Epub 2014 Jul 15. Arbizu J, Luquin MR, Abella J, de la Fuente-Fernandez R, Fernandez-Torron R, Garcia-Solis D, et al. Functional neuroimaging in the diagnosis of patients with Parkinsonism: Update and recommendations for clinical use. Revista espanola de medicina nuclear e imagen molecular. 2014;33(4): Epub 2014 Apr 14. Narvaez M, Millon C, Borroto-Escuela D, Flores-Burgess A, Santin L, Parrado C, et al. Galanin receptor 2-neuropeptide Y Y1 receptor interactions in the amygdala lead to increased anxiolytic actions. Brain structure & function. [Epub ahead of print]. Epub 2014 May 20. Gonzalez-Redondo R, Garcia-Garcia D, Clavero P, Gasca-Salas C, Garcia-Eulate R, Zubieta JL, et al. Grey matter hypometabolism and atrophy in Parkinson's disease with cognitive impairment: a twostep process. Brain: a journal of neurology. 2014;137(Pt 8): Epub 2014 Jun 20. Agalliu I, San Luciano M, Mirelman A, Giladi N, Waro B, Aasly J, et al. Higher Frequency of Certain Cancers in LRRK2 G2019S Mutation Carriers With Parkinson Disease: A Pooled Analysis. JAMA neurology. 2015;72(1): Epub Lemmers RJ, Goeman JJ, van der Vliet PJ, van Nieuwenhuizen MP, Balog J, Vos-Versteeg M, et al. Inter-individual differences in CpG methylation at D4Z4 correlate with clinical variability in FSHD1 and FSHD2. Human molecular genetics. 2015;24(3): Epub 2014 Sep 25. Gasca-Salas C, Estanga A, Clavero P, Aguilar-Palacio I, Gonzalez-Redondo R, Obeso JA, et al. Longitudinal assessment of the pattern of cognitive decline in non-demented patients with advanced Parkinson's disease. Journal of Parkinson's disease. 2014;4(4): Gomez-Suaga P, Rivero-Rios P, Fdez E, Blanca Ramirez M, Ferrer I, Aiastui A, et al. LRRK2 delays degradative receptor trafficking by impeding late endosomal budding through decreasing Rab7 activity. Human molecular genetics. 2014;23(25): Epub 2014 Jul 30. Etxaniz U, Perez-San Vicente A, Gago-Lopez N, Garcia-Dominguez M, Iribar H, Aduriz A, et al. Neural-competent cells of adult human dermis belong to the Schwann lineage. Stem cell reports. 2014;3(5): Epub 2014 Oct 16. Marti Masso JF, Zarranz JJ, Otaegui D, de Munain AL. Neurogenetic disorders in the basque population. Annals of human genetics. 2015;79(1): Epub 2014 Dec 1. Gomez-Uriz AM, Milagro FI, Mansego ML, Cordero P, Abete I, De Arce A, et al. Obesity and ischemic stroke modulate the methylation levels of KCNQ1 in white blood cells. Human molecular genetics. [Epub ahead of print]. Epub 2014 Nov 4. Geurtsen GJ, Hoogland J, Goldman JG, Schmand BA, Tröster AI, Burn DJ, et al. Parkinson's disease mild cognitive impairment: application and validation of the criteria. Journal of Parkinson's disease. 2014;4(2): Obeso JA, Rodriguez-Oroz MC, Stamelou M, Bhatia KP, Burn DJ. The expanding universe of disorders of the basal ganglia. Lancet. 2014;384(9942): Epub 2014 Jun 18. Program 3 244

251 2014 Annual Report Adolfo López de Munain Group Obeso I, Wilkinson L, Casabona E, Speekenbrink M, Luisa Bringas M, Alvarez M, et al. The subthalamic nucleus and inhibitory control: impact of subthalamotomy in Parkinson's disease. Brain: a journal of neurology. 2014;137(Pt 5): Epub 2014 Mar 22. Hallast P, Batini C, Zadik D, Maisano Delser P, Wetton JH, Arroyo-Pardo E, et al. The Y-Chromosome Tree Bursts into Leaf: 13,000 High-Confidence SNPs Covering the Majority of Known Clades. Molecular biology and evolution. 2014;32(3): Epub 2014 Dec 2. Höglinger GU, Huppertz HJ, Wagenpfeil S, Andres MV, Belloch V, Leon T, et al. Tideglusib reduces progression of brain atrophy in progressive supranuclear palsy in a randomized trial. Movement disorders: official journal of the Movement Disorder Society. 2014;29(4): Epub 2014 Jan 31. Irizar H, Munoz-Culla M, Sepulveda L, Saenz-Cuesta M, Prada A, Castillo-Trivino T, et al. Transcriptomic profile reveals gender-specific molecular mechanisms driving multiple sclerosis progression. PloS one. 2014;9(2):e Epub 2014 Feb 28. Martinez-Martin P, Hernandez B, Ricart J; FAST Study Group. Factors determining when to start levodopa/carbidopa/entacapone treatment in Spanish patients with Parkinson's disease. Neurologia (Barcelona, Spain). 2014;29(3): Alquezar C, Esteras N, de la Encarnacion A, Alzualde A, Moreno F, Lopez de Munain A, et al. PGRN haploinsufficiency increased Wnt5a signaling in peripheral cells from frontotemporal lobar degeneration-progranulin mutation carriers. Neurobiology of aging. 2014;35(4): Toledo JB, Lopez-Azcarate J, Garcia-Garcia D, Guridi J, Valencia M, Artieda J, et al. High beta activity in the subthalamic nucleus and freezing of gait in Parkinson's disease. Neurobiology of disease. 2014;64:60-5. Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodriguez-Rodriguez E, Lopez de Munain A, et al. Assessing the role of the TREM2 p.r47h variant as a risk factor for Alzheimer's disease and frontotemporal dementia. Neurobiology of aging. 2014;35(2):444.e1-4. Tercjak A, Bergareche A, Caballero C, Tunon T, Linazasoro G. Lewy bodies under atomic force microscope. Ultrastructural pathology. 2014;38(1):1-5. Garcia-Parra P, Naldaiz-Gastesi N, Maroto M, Padin JF, Goicoechea M, Aiastui A, et al. Murine muscle engineered from dermal precursors: an in vitro model for skeletal muscle generation, degeneration, and fatty infiltration. Tissue engineering. Part C, Methods. 2014;20(1): Martinez-Martin P, Rodriguez-Blazquez C, Forjaz MJ, Alvarez-Sanchez M, Arakaki T, Bergareche-Yarza A, et al. Relationship between the MDS-UPDRS domains and the health-related quality of life of Parkinson's disease patients. European journal of neurology: the official journal of the European Federation of Neurological Societies. 2014;21(3): Riancho-Zarrabeitia L, Delgado-Alvarado M, Riancho J, Oterino A, Sedano MJ, Rueda-Gotor J, et al. Anti-TNF-α therapy in the management of severe neurosarcoidosis: a report of five cases from a single centre and literature review. Clinical and experimental rheumatology. 2014;32(2): Ruiz-Martinez J, de la Riva P, Rodriguez-Oroz MC, Mondragon Rezola E, Bergareche A, Gorostidi A, et al. Prevalence of cancer in Parkinson's disease related to R1441G and G2019S mutations in LRRK2. Movement disorders: official journal of the Movement Disorder Society. 2014;29(6): Lopez de Maturana R, Aguila JC, Sousa A, Vazquez N, Del Rio P, Aiastui A, et al. Leucine-rich repeat kinase 2 modulates cyclooxygenase 2 and the inflammatory response in idiopathic and genetic Parkinson's disease. Neurobiology of aging. 2014;35(5): Delgado-Alvarado M, Gomez-Roman J, Sanchez-Salmon E, Rodriguez-Rodriguez E, Polo JM, Garcia- Castano A, et al. Nonanaplastic pleomorphic xanthoastrocytoma with meningeal dissemination presenting with bilateral visual loss. Journal of neuroimaging: official journal of the American Society of Neuroimaging. 2014;24(5):

252 Research Projects Code: PI11/03028 Title: DEMTEST: Biomarker based diagnosis of rapid progressive dementias Principal Investigator: Pascual Sánchez CIBERNED s collaboration: Yes Groups CIBERNED: G601; G114; G503; G609 Type: Others Funding Agency: Instituto de Salud Carlos III - Comunidad Económica Europea Funding: Duration: Code: PI13/00722 Title: Análisis in vitro del recambio (turnover) proteico en músculo de pacientes con distrofia de cinturas tipo 2A Principal Investigator: Amets Saenz Groups CIBERNED: Type: National Funding Agency: FIS Funding: Duration: 2014 Code: BECAGVHARITZ Title: Beca formación investigador-modalidad predoctoral(a) haritz jimenez urbieta Principal Investigator: Maria Cruz Rodríguez Oroz Groups CIBERNED: Tipo: CCAA Funding Agency: Departamento de Educación, Politica Lingüística y Cultura Funding: Duration: 2014 Code: GANGOITIMANUELDELGADO Title: Biomarcadores de deterioro cognitivo leve en enfermedad de Parkinson Principal Investigator: Maria Cruz Rodríguez Oroz Groups CIBERNED: Type: International Funding Agency: GANGOITI Funding: Duration: 2014 Code: GV Title: Estudio de Biomarcadores en la fase temprana de la demencia en la enfermedad de Parkinson Principal Investigator: María Cruz Rodríguez Oroz Groups CIBERNED: Type: International Funding Agency: Gobierno Vasco Funding: Duration: Code: MIRAMAR Title: Factores neuroprotectores de demencia en una muestra de nonagenarios cognitivamente sanos con depósitos de betaamiloide cerebral comparada con pacientes con enfermedad de Alzheimer (Proyecto Miramar) Principal Investigator: Adolfo López de Munain Groups CIBERNED: Type: International Funding Agency: Fundación 2000 Funding: Duration: 2014 Code: SAIO13-PE13BN012 Title: Mecanismos fisiopatológicos del trastorno del control de impulsos en la enfermedad de parkinson Principal Investigator: Maria Cruz Rodríguez Oroz Groups CIBERNED: Type: National Funding Agency: Departamento de Desarrollo Económico y Competitividad Funding: Duration: 2014 Code: MULTIPAGEN Title: Multimodal MRI in Basque LRRK2 PD. Multipagen Principal Investigator: Maria Cruz Rodríguez Oroz Groups CIBERNED: Type: National Funding Agency: Red transfronteriza de investigación biomédica de los pirineos Funding: Duration: 2014 Code: DPPE14/001 Title: Testado de nuevas sustancias con potencial terapéutico in vitro e in vivo para el tratamiento de la distro- Program 3 246

253 2014 Annual Report Adolfo López de Munain Group fia muscular de Duchenne Principal Investigator: Ainara Vallejo Groups CIBERNED: Type: International Funding Agency: Asociación Duchenne Parent Project España Funding: Duration: PhD Dissertations Author: Oihane Jaka Irizar Title: LGMD2A gerrietako muskulu-distrofiaren adierazpen-analisiaren aplikazioa gaixotasunaren diagnostikoan, eta sagu-ereduan eta giza eredu zelularretan oinarritutako karakterizazio fisiopatologikoan Date: 21st March 2014 Supervisor: Amets Sáenz Peña 247

254 GROUP Pura Muñoz Cánoves Program 3 CONTACT DETAILS oup Pura Muñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves Cell Group Biology Pura Group Muñoz Cánoves ra Muñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz ICREA and Cánoves Pompeu Group Fabra Pura University Muñoz (UPF) Cánoves Group a Department of Experimental and Life Sciences (DCEXS) uñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves Group Phone: Pura +34 Muñoz Cánoves 891 Group Pura Fax: s Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves pura.munoz@upf.edu oup Pura Muñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves Group Pura Muñoz Cánoves LIST OF PERSONNEL Pura Muñoz Cánoves Principal Investigator, ICREA Research Professor Antonio L. Serrano Sánchez Senior Researcher Eusebio Perdiguero Santamaría Senior Researcher CIBERNED Laura García Prat PhD student Susana Gutarra Díaz Technician Mercè Jardí Ripoll Technician Yacine Kharraz Postdoctoral Fellow Vera Lukesova Technician Laura Ortet Postdoctoral Fellow Patrizia Pessina Postdoctoral Fellow Marina Raya Labmanager Vanessa Ruíz-Bonilla Postdoctoral Fellow Mónica Zamora Villafaina Postdoctoral Fellow Jessica Segales Dalmau Postdoctoral Fellow Diana Sobral Mesquita Predoctoral Fellow Begoña Ampudia Carrasco Technician Beatriz de Lucas Moreno Predoctoral Fellow 248

255 2014 Annual Report Pura Muñoz Cánoves Group Summary Regulation of skeletal myogenesis by p38 MAPK signaling during myogenesis, at the different stages of the in vivo regeneration process: activation, proliferation, differentiation and return to quiescence (selfrenewal). New roles for the p38/mkp-1 balance in controlling macrophage phenotypic transitions necessary for appropriate muscle stem cell dependent tissue repair. This novel regulatory mechanism may have implications in chronic inflammatory degenerative diseases. Matrix-associated fibrin-driven inflammation coupled to the αmβ2 leukocyte integrin receptor modulates muscle regeneration and/or fibrosis. We found that fibrin accumulates in dystrophic muscles of DMD patients and mdx mice and disruption of fibrin-αmβ2 interactions attenuated dystrophy progression. Identification of the extracellular upa/pai-1 proteolytic balance as an important regulator of mir 21 biogenesis, controlling age-associated muscle fibrosis and dystrophy progression. Fibrosis in aged mdx mice could be reversed by mir-21 or upa-selective interference. Regulation of muscle growth by IL-6. Our results have provided novel functional evidence for the critical involvement of the cytokine IL-6 in mediating hypertrophic muscle growth by controlling muscle stem cell (satellite cell) incorporation to growing myofibers. Geriatric muscle stem cells switch reversible quiescence into senescence, due to derepression of p16ink4a. We demonstrated that p16ink4a silencing in geriatric satellite cells restores quiescence and muscle regenerative functions. This may provide a basis for stem cell rejuvenation in sarcopenic muscles. Keywords Skeletal muscle, Regeneration, Inflammation, Fibrosis, Muscle stem cells, Sarcopenia, Aging Publications Lopez-Arribillaga E, Rodilla V, Pellegrinet L, Guiu J, Iglesias M, Roman AC, et al. Bmi1 regulates murine intestinal stem cell proliferation and self-renewal downstream of Notch. Development (Cambridge, England). 2015;142(1): Epub 2014 Dec 5. Segales J, Perdiguero E, Munoz-Canoves P. Epigenetic control of adult skeletal muscle stem cell functions. The FEBS journal. [Epub ahead of print]. Epub 2014 Sep 22. Sousa-Victor P, Gutarra S, Garcia-Prat L, Rodriguez-Ubreva J, Ortet L, Ruiz-Bonilla V, et al. Geriatric muscle stem cells switch reversible quiescence into senescence. Nature. 2014;506(7488): Epub 2014 Feb 12. Sousa-Victor P, Perdiguero E, Munoz-Canoves P. Geroconversion of aged muscle stem cells under regenerative pressure. Cell cycle (Georgetown, Tex.). 2014;13(20): Bouche M, Munoz-Canoves P, Rossi F, Coletti D. Inflammation in muscle repair, aging, and myopathies. 2014;2014: Epub 2014 Aug 4. Pessina P, Cabrera D, Morales MG, Riquelme CA, Gutierrez J, Serrano AL, et al. Novel and optimized strategies for inducing fibrosis in vivo: focus on Duchenne Muscular Dystrophy. Skeletal muscle. 2014;4:7. Epub 2014 Aug

256 Kharraz Y, Guerra J, Pessina P, Serrano AL, Munoz-Canoves P. Understanding the process of fibrosis in Duchenne muscular dystrophy. Biomed Research International. 2014;2014: Epub 2014 May 4. Acuna MJ, Pessina P, Olguin H, Cabrera D, Vio CP, Bader M, et al. Restoration of muscle strength in dystrophic muscle by angiotensin-1-7 through inhibition of TGF-β signalling. Human molecular genetics. 2014;23(5): Research Projects Code: DPP Title: Controlling fibrosis for improving muscle recovery and cell therapy efficiency in DMD Principal Investigator: Pura Muñoz Cánoves Groups CIBERNED: Type: International Funding Agency: DPP NL Funding: Duration: 2014 Code: ENDOSTEM Title: ENDOSTEM Optimization of stem cell therapy for disease of epithelia and skeletal muscle through combined basic and applied research Principal Investigator: David Sassoon Groups CIBERNED: Type: European Funding Agency: UE Funding: Duration: Code: La Marató - TV3 Title: Novel approaches for degenerative muscular dystrophies therapies Principal Investigator: Pura Muñoz Cánoves Groups CIBERNED: Type: National Funding Agency: La Marató - TV3 Funding: Duration: Code: SAF Title: Regulation and dysregulation of skeletal muscle regeneration and growth Principal Investigator: Pura Muñoz Cánoves Groups CIBERNED: Type: National Funding Agency: MINECO Funding: Duration: Code: ERARE13-fp-149 Title: Stimulating Intrinsic Repair for DMD (SIRD) Principal Investigator: Michael Rudnicki Groups CIBERNED: Type: International Funding Agency: CIBERNED Funding: Duration: 2014 Code: Inter-CIBERs Title: Molecular links between diabetes and neurodegenerative disorders Principal Investigator: Angel Raya CIBERNED s collaboration: Yes Groups CIBERNED: G409; G604 Other CIBER s collaboration: Yes (CIBERDEM) Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: 2014 Program 3 250

257 GROUP Xavier Navarro Acebes Program 3 CONTACT DETAILS Group Xavier Navarro Acebes Group Xavier Navarro Dept. Acebes Cell Group Biology, Xavier Physiology Navarro and Acebes Immunology Group Xavier Navarr Acebes Group Xavier Navarro Acebes Group Xavier Navarro Acebes Group Institute Xavier of Navarro Neurosciences Acebes Group Xavie Navarro Acebes Group Xavier Navarro Acebes Group Xavier Navarro Universitat Acebes Autonoma Group Xavier de Barcelona Navarro Acebes Grou Edif. M, Campus UAB Xavier Navarro Acebes Group Xavier Navarro Acebes Group Xavier Navarro Acebes Bellaterra Group (Spain) Xavier Navarro Acebe Group Xavier Navarro Acebes Group Xavier Navarro Acebes Group Xavier Phone: Navarro +34 Acebes Group 966 Xavier Navarr Fax: Acebes Group Xavier Navarro Acebes Group Xavier Navarro Acebes Group xavier.navarro@uab.cat Xavier Navarro Acebes Group Xavie Navarro Acebes Group Xavier Navarro Acebes Group Xavier Navarro Acebes Group Xavier Navarro Acebes Group LIST OF PERSONNEL Xavier Navarro Acebes Principal Investigator, Full Professor Jordi Cuadras Mas Researcher, Emeritous Professor Esther Udina Bonet Researcher, Associate Professor Rubèn López Vales Researcher, Associate Professor interim Caty Casas Louzao Researcher, Associate Professor interim Assumpció Bosch Merino Researcher, Associate Professor Jordi Bruna Escuer Assistant Professor Stefano Cobianchi Postdoctoral Fellow Jaume del Valle Macià Postdoctoral Fellow Joaquim Hernández Martín Postdoctoral Fellow Mireia Herrando Grabulosa Postdoctoral Fellow Albert Alé Miranda PhD student Ariadna Arbat Plana PhD student Marina Coll Miró PhD student Natalia De la Oliva Muñoz PhD student Isaac Francos Quijorna PhD student Francisco González Pérez PhD student Víctor Manuel López Álvarez PhD student Clara López Serrano PhD student Renzo Mancuso PhD student Anna Martínez Muriana PhD student Elena Redondo Castro PhD student David Romeo Guitart PhD student Eva Santos Nogueira PhD student Daniel Santos Rojas PhD student Abel Torres Espín PhD student Jordi Badia Casahuja Researcher Jessica Jaramillo Rodríguez Technician Marta Morell Orduña Technician 251

258 Summary The Group of Neuroplasticity and Regeneration of the UAB is a multidisciplinary research group working on repair, regeneration and functional recovery after peripheral nerve and spinal cord lesions and in neurodegenerative diseases. The research activities of the Group of Neuroplasticity and Regeneration have focused on the study of physiopathological mechanisms of neural lesions, neuropathic pain and neurodegeneration, and on the application of novel therapeutic strategies for regenerating traumatic and degenerative lesions of the nervous system. The active research lines include: Cellular and molecular mechanisms implicated in degeneration of motoneurons in experimental models. Search for biomarkers. Neuroprotective strategies based on gene and pharmacological therapy. Cell therapy by transplantation of mesenchimal stem cells and olfactory glia for the repair of spinal cord injuries and motoneuron degenerative diseases. Repair of spinal root avulsion injuries by surgical reimplantation and pharmacological neuroprotection. Etiopathogenic role of lipid mediators and modulators of the neuroinflammatory response in neurodegeneration induced by central nervous system lesions. Activity-dependent therapies for enhancing axonal regeneration and functional recovery after peripheral nerve lesions. Physiopathologic mechanisms in neuropathic pain induced by nerve and spinal cord lesions. Study of the relationship between neuroinflammation and hyperexcitability. Study of etiopathogenic mechanisms and potential neuroprotective treatments in peripheral neuropathies induced by diabetes and by antitumoral agents. Design and evaluation of neural interfaces for the development of neuroprostheses applied to neurorehabilitation. Study of new intraneural electrodes for the selective stimulation and recording of neural activity. Keywords Neurodegeneration, Nerve regeneration, Spinal cord injury, Neuropathic pain, Motoneuron diseases, Peripheral neuropathies, Neural interfaces Publications Arbat-Plana A, Torres-Espin A, Navarro X, Udina E. Activity dependent therapies modulate the spinal changes that motoneurons suffer after a peripheral nerve injury. Experimental neurology. 2015;263: Epub 2014 Oct 23. Cobianchi S, de Cruz J, Navarro X. Assessment of sensory thresholds and nociceptive fiber growth after sciatic nerve injury reveals the differential contribution of collateral reinnervation and nerve regeneration to neuropathic pain. Experimental neurology. 2014;255:1-11. Epub 2014 Feb 16. Program 3 252

259 2014 Annual Report Xavier Navarro Acebes Group Torres-Espin A, Redondo-Castro E, Hernandez J, Navarro X. Bone marrow mesenchymal stromal cells and olfactory ensheathing cells transplantation after spinal cord injury - a morphological and functional comparison in rats. The European journal of neuroscience. 2014;39(10): Epub 2014 Mar 18. Mòdol L, Mancuso R, Ale A, Francos-Quijorna I, Navarro X. Differential effects on KCC2 expression and spasticity of ALS and traumatic injuries to motoneurons. Frontiers in cellular neuroscience. 2014;8:7. Epub 2014 Jan 24. Ariza L, Pagès G, Garcia-Lareu B, Cobianchi S, Otaegui PJ, Ruberte J, et al. Experimental diabetes in neonatal mice induces early peripheral sensorimotor neuropathy. Neuroscience. 2014;274: Epub 2014 May 15. Brunner SM, Farzi A, Locker F, Holub BS, Drexel M, Reichmann F, et al. GAL3 receptor KO mice exhibit an anxiety-like phenotype. Proceedings of the National Academy of Sciences of the United States of America. 2014;111(19): Epub 2014 Apr 29. Torres-Espin A, Redondo Castro E, Hernandez J, Navarro X. Immunosuppression of allogenic mesenchymal stem cells transplantation after spinal cord injury improves graft survival and beneficial outcomes. Journal of neurotrauma. [Epub ahead of print]. Epub 2014 Sep 9. Homs J, Pages G, Ariza L, Casas C, Chillon M, Navarro X, et al. Intrathecal administration of IGF-I by AAVrh10 improves sensory and motor deficits in a mouse model of diabetic neuropathy. Molecular Therapy - Methods & Clinical Development. 2014;1:7. Mancuso R, Del Valle J, Morell M, Pallas M, Osta R, Navarro X. Lack of synergistic effect of resveratrol and sigma-1 receptor agonist (PRE-084) in SOD1 G93A ALS mice: overlapping effects or limited therapeutic opportunity? Orphanet journal of rare diseases. 2014;9:78. Epub 2014 May 21. David M, Machuca-Gayet I, Kikuta J, Ottewell P, Mima F, Leblanc R, et al. Lysophosphatidic acid receptor type 1 (LPA1) plays a functional role in osteoclast differentiation and bone resorption activity. The Journal of biological chemistry. 2014;289(10): Epub 2014 Jan 15. Darbra S, Mòdol L, Llido A, Casas C, Vallee M, Pallarès M. Neonatal allopregnanolone levels alteration: effects on behavior and role of the hippocampus. Progress in neurobiology. 2014;113: Epub 2013 Aug 17. Mòdol L, Casas C, Llido A, Navarro X, Pallarès M, Darbra S. Neonatal allopregnanolone or finasteride administration modifies hippocampal K(+) Cl(-) co-transporter expression during early development in male rats. The Journal of steroid biochemistry and molecular biology. 2014;143: Epub 2014 May 23. Torres-Espin A, Santos D, Gonzalez-Perez F, del Valle J, Navarro X. Neurite-J: an image-j plug-in for axonal growth analysis in organotypic cultures. Journal of neuroscience methods. 2014;236: Epub 2014 Aug 12. Ale A, Bruna J, Navarro X, Udina E. Neurotoxicity induced by antineoplastic proteasome inhibitors. Neurotoxicology. 2014;43: Epub 2014 Feb 10. Giudetti G, Del Valle Macia J, Navarro Acebes X, Micera S. NGF-loaded PLGA microparticles for advanced multifunctional regenerative electrodes. Conference proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference. 2014;2014: Mancuso R, Osta R, Navarro X. Presymptomatic electrophysiological tests predict clinical onset and survival in SOD1(G93A) ALS mice. Muscle & nerve. 2014;50(6): Epub 2014 Oct 30. Mòdol L, Cobianchi S, Navarro X. Prevention of NKCC1 phosphorylation avoids downregulation of KCC2 in central sensory pathways and reduces neuropathic pain after peripheral nerve injury. Pain. 2014;155(8): Epub 2014 May 9. Navarro X, Del Valle J. Regenerative neural interfaces for neuroprosthetic applications. Front Neuroeng. 2014; Conference Abstract: MERIDIAN 30M Workshop. Velasco R, Videla S, Villoria J, Ortiz E, Navarro X, Bruna J. Reliability and accuracy of quantitative sensory testing for oxaliplatin-induced neurotoxicity. Acta neurologica Scandinavica. [Epub ahead of print]. Epub 2014 Oct

260 Mancuso R, del Valle J, Modol L, Martinez A, Granado-Serrano AB, Ramirez-Nunez O, et al. Resveratrol improves motoneuron function and extends survival in SOD1(G93A) ALS mice. Neurotherapeutics: the journal of the American Society for Experimental NeuroTherapeutics. 2014;11(2): Allodi I, Mecollari V, Gonzalez-Perez F, Eggers R, Hoyng S, Verhaagen J, et al. Schwann cells transduced with a lentiviral vector encoding Fgf-2 promote motor neuron regeneration following sciatic nerve injury. Glia. 2014;62(10): Epub 2014 Jul 2. Gonzalez-Perez F, Cobianchi S, Geuna S, Barwig C, Freier T, Udina E, et al. Tubulization with chitosan guides for the repair of long gap peripheral nerve injury in the rat. Microsurgery. [Epub ahead of print]. Epub 2014 Dec 4. Modol L, Casas C, Navarro X, Llido A, Vallee M, Pallarès M, et al. Neonatal finasteride administration alters hippocampal α4 and δ GABAAR subunits expression and behavioural responses to progesterone in adult rats. The international journal of neuropsychopharmacology: official scientific journal of the Collegium Internationale Neuropsychopharmacologicum (CINP). 2014;17(2): Redondo-Castro E, Navarro X. Chronic ibuprofen administration reduces neuropathic pain but does not exert neuroprotection after spinal cord injury in adult rats. Experimental neurology. 2014;252: Ale A, Bruna J, Morell M, van de Velde H, Monbaliu J, Navarro X, et al. Treatment with anti-tnf alpha protects against the neuropathy induced by the proteasome inhibitor bortezomib in a mouse model. Experimental neurology. 2014;253: Research Projects Code: FP Title: Biohybrid templates for peripheral nerve regeneration (BIOHYBRID) Principal Investigator: Xavier Navarro Acebes Groups CIBERNED: Type: European Funding Agency: European Commission Funding: Duration: Code: FP7-NMP4-SL Title: Micro and nano engineered bidirectional carbon interfaces for advanced peripheral nervous system prosthetic and hybrid bionics (MERIDIAN) Principal Investigator: Xavier Navarro Acebes Groups CIBERNED: Type: International Funding Agency: European Commission Funding: Duration: Code: PI11/00464 Title: Modulación del arco reflejo de estiramiento para mejorar la recuperación funcional despues de lesiones nerviosas Principal Investigator: Esther Udina Bonet Groups CIBERNED: Type: National Funding Agency: Fondo de Investigación Sanitaria, Instituto Salud Carlos III Funding: Duration: Code: FP Title: Natural sensory feedback for phantom limb pain modulation and therapy (EPIONE) Principal Investigator: Xavier Navarro Acebes Groups CIBERNED: Type: International Funding Agency: Project FP Funding: Duration: Code: FP Title: Neurocontrolled bidirectional artificial upper limb and hand prosthesis (NEBIAS) Principal Investigator: Xavier Navarro Acebes Groups CIBERNED: Type: International Funding Agency: European Commission Funding: Duration: Program 3 254

261 2014 Annual Report Xavier Navarro Acebes Group Code: SAF R Title: Participación de los receptores del ácido lisofosfatídico en la fisiopatología de la lesión medular Principal Investigator: Rubèn López Vales Groups CIBERNED: Type: International Funding Agency: Ministerio de Economía y Competitividad Funding: Duration: Code: TV Title: Potenciación de los mecanismos endógenos de neuroprotección y reparación del sistema nervioso despues de daño agudo Principal Investigator: Caty Casas Groups CIBERNED: Other CIBER s collaboration: Yes (CIBERER) Type: National Funding Agency: Fundació Marató TV3 Funding: Duration: Code: RD12/0019/0011 Title: RETIC de Terapia Celular Principal Investigator: Xavier Navarro Acebes (Coordinador de red: José María Moraleda) CIBERNED s collaboration: Yes Groups CIBERNED: G301; G102; G113; G208; G105; G607 Type: National Funding Agency: Instituto de Salud Carlos III Funding: Duration: Code: PIRG05-GA Title: Role of lysophosphatidic acid in the pathophysiology of spinal cord injury (ROLPASCI) Principal Investigator: Xavier Navarro Acebes Groups CIBERNED: Type: European Funding Agency: Marie-Curie International Reintegration Grant, European Commission FP7 Funding: Duration: Code: IRP Research Grant P148 Title: Role of pro-resolving lipid mediators in SCI Principal Investigator: Rubèn López Vales Groups CIBERNED: Type: International Funding Agency: International Foundation for Research in Paraplegia Funding: Duration: Code: TV Title: Therapeutic role of IL-37 in spinal cord injury Principal Investigator: Rubèn López Vales Groups CIBERNED: Type: National Funding Agency: Fundació Marató TV3 Funding: Duration: PhD Dissertations Author: Renzo Mancuso Title: Electrophysiological biomarkers for the study of new therapeutic strategies for motoneuron disease Date: 18th June 2014 Supervisor: Xavier Navarro Acebes Author: Albert Alé Miranda Title: Neurotoxicity induced by proteasome inhibitor bortezomib. Study of the mechanisms of action and evaluation of neuroprotective strategies in a mouse model Date: 12nd December 2014 Supervisor: Esther Udina Bonet Author: Eva Santos Nogueira Title: Spinal cord injury. Role of endothelial differentiation gene family lysophosphatidic acid receptors Date: 18th December 2014 Supervisor: Rubèn López Vales 255

262 GROUP Juan Jesús Vílchez Padilla Program 3 CONTACT DETAILS oup Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Padilla Hospital Group Universitari Juan Jesús Vílchez i Politecnic Padilla La Group Fe Juan Jesús lchez Padilla Group Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Padilla Servicio Group de Neurología Juan Jesús Vílchez Padilla oup Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Torre Padilla D, planta Group 5 Juan Jesús Bulevar Sur, s/n lchez Padilla Group Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Padilla Group Valencia Juan (Spain) Jesús Vílchez Padilla oup Juan Jesús Vílchez Padilla Group Phone: +34 Juan 961 Jesús 244 Vílchez 000 (centralita) Padilla Group / Juan 543 Jesús (secretaria Vílchez neurología) Padilla Group Juan Jesús vilchez_jje@gva.es lchez Padilla Group Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Padilla Group Juan Jesús Vílchez Padilla G LIST OF PERSONNEL Juan Jesús Vílchez Padilla Principal Investigator, Associate Professor, Chief of Neurology Department Teresa Sevilla Mantecón Neurologist Luís Bataller Alberola Neurologist María José Chumillas Clinical Neurophysiologist Nuria Muelas Gómez PhD, Neurologist Rafael Sivera Neurologist Juan Francisco Vazquez Costa Post-MIR Fernándo Mayordomo Researcher, Morphology and Neuromuscular Pathology Inmaculada Azorín Villena CIBERNED Postdoctoral Fellow Pilar Martí PhD student E. Torres Vega PhD student Clara Gomis Coloma PhD student Lorena Perpiñá Gómez CIBERNED Technician Roger Vílchez Medina Lab Technician 256

263 2014 Annual Report Juan Jesús Vílchez Padilla Group Summary In the area of genetic neuropathies, particularly in Charcot Marie Tooth Disease (CMT) neuropathies, in the annuity 2013, it has been established the constitution of clinical Workpage led by Dr. Sevilla within the cooperative project TREAT- CMT who was elected for funding by the European call ISCIII-IRDYC Among his achievements is the creation of a unified database which contains a casuistry of CMT over 700 patients in which prominent Spanish centers (La Paz, Bellvitge, Rocío, etc.) has been involved. As an example of this work has been published a series CMT full genetic characterization which is a reference in this field. Other contributions of interest are clinical and genetic reassessment of hereditary motor and sensory neuropathy type Russe associated with a founder mutation affecting a predominantly Spanish Gypsies, also has published a study original on the vestibular involvement as a primary symptom in CMT4A neuropathy (Perez-Garrigues et al., 2014). A milestone in the field of motor neuron disease, also led by Dr. Sevilla, it has been obtaining a fellowship in Post- Mir public call by the ISS La Fe awarded to Juan Francisco Vazquez Costa, on a research oriented ELA project, developed in collaboration with the group of Dr. Neurogenetic Jordi Pérez-Tur, Institute of Biomedicine of Valencia. In the field of congenital myopathies and myasthenias, led by Dr. Muelas and Dr. Vilchez, the highlights were our participation in the discovery of a mutation in the gene that is responsible Transportina III Muscular Dystrophy LGMD1F in collaboration with several Spanish and international groups. This form of dystrophy is unique to a Spanish family whose clinical following up is mainly performed in our hospital, which has allowed the publication of additional data on the clinical phenotype in collaboration with Italian researchers, and also it has put forward the basis for undertaking projects with prestigious groups in order to decipher the pathogenesis and treatment of this myopathy and to deepen transport in the pathogenesis of HIV virus that depends on the Transportina III as well. Another notable collaboration is the study of mirna dysregulation in a model of both Drosophila and human biopsies of patients with myotonic dystrophy has clarified that data on the pathogenesis of the disease. In the field of neuromuscular diseases and autoimmune encephalopathies led by Dr. Bataller should be noted the confirmation of the presence of ACS against aquaporin serum of patients with paraneoplastic myelitis-hyperckemia as expression of an ovarian teratoma. We also noticed two major collaborations on the involvement of NMDA receptor Abs against both pediatric encephalitis and late encephalitis. This field should also be noted obtaining a predoctoral fellowship EAI (Valencian Health Department) for a degree in Biology (E. Torres Vega) under the direction of Dr. Bataller develop a project to decipher the pathogenesis of the thymus in developing neuromyotonia, myasthenia syndromes. Keywords Charcot-Marie-Tooth disease, Genetic neuropathy type Russe, CMT4G, HK1 gene, CMT4A, SH3-TCH gene, Vestibular neuropathy, LGMD1F, Transportina III, Myotonic dystrophy, CTG expansion, mirna, Limbic encephalitis, Myelitis, Anti-NMDA receptor Abs, Anti-Aquoporin Abs 257

264 Publications Vazquez-Costa JF, Baquero-Toledo M, Sastre-Bataller I, Mas-Estelles F, Vilchez-Padilla JJ. Inflammatory amyloid angiopathy. Neurologia (Barcelona, Spain). 2014;29(4): Epub 2012 Aug 11. Lamont PJ, Wallefeld W, Hilton-Jones D, Udd B, Argov Z, Barboi AC, et al. Novel mutations widen the phenotypic spectrum of slow skeletal/β-cardiac myosin (MYH7) distal myopathy. Human mutation. 2014;35(7): Epub 2014 May 21. Kornak U, Mademan I, Schinke M, Voigt M, Krawitz P, Hecht J, et al. Sensory neuropathy with bone destruction due to a mutation in the membrane-shaping atlastin GTPase 3. Brain: a journal of neurology. 2014;137(Pt 3): Epub 2014 Jan 22. Perez-Garrigues H, Sivera R, Vilchez JJ, Espinos C, Palau F, Sevilla T. Vestibular impairment in Charcot- Marie-Tooth disease type 4C. Journal of neurology, neurosurgery, and psychiatry. 2014;85(7): Epub 2014 Mar 10. Research Projects Code: Proyecto de Investigación Terapia en Distrofias Musculares / Fundación Isabel Gemio Title: Biología, fisiopatología y terapia de las células satélite del músculo esquelético Principal Investigator: Juan J Vílchez Padilla Groups CIBERNED: Type: Private Funding Agency: Fundación Isabel Gemio Funding: Duration: Code: ACIF/2012/047 Title: Estudio de autoinmunidad en síndromes neuromusculares asociados a timoma Principal Investigator: Luis Bataller Alberola Groups CIBERNED: Tipo: CCAA Funding Agency: Generalitat Valenciana Funding: Duration: 2014 Code: PI11/01330 Title: Estudio de la Hiperckemia Asintomática o Paucisintomática Principal Investigator: Juan J Vílchez Groups CIBERNED: Type: National Funding Agency: FIS Funding: Duration: Code: 2013_0312_CPC_POSTRESIDENTES Title: Estudio integral de la Esclerosis Lateral Amiotrófica en una Unidad de Referencia Principal Investigator: Teresa Sevilla Groups CIBERNED: Type: International Funding Agency: Instituto de Investigación Sanitaria La Fe Funding: N.D. Duration: 2014 Program 3 258

265 Cooperative Research COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE PERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESE TIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARC RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH SEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPE COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERA COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE PERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RE RATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESE TIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARC RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH COOPERATIVE RESEARCH C

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267 2014 Annual Report Cooperative Research The cooperative research program initiated in 2010 continues to be viewed as an essential scientific tool for planning CIBERNED future actions. This program is one of the cornerstones of the Centre s activities, and aims to encourage collaborative research between the various research groups, joining forces and exploiting synergies and complementary skills. It is intended to identify collaborative research projects with a marked innovative and translational nature where the cooperative effort significantly increase the added value of the research activity. The proposals, although subject to final approval by the Steering Committee, are externally reviewed by the National Evaluation Agency (ANEP), so that the allocation of funds is independent and transparent. The cooperative research program has so far launched a total of four calls for projects. The first two (2010 and 2011) have been already completed with a total of ten approved projects and a total accumulated budget allocation of 3,514,100 Euros. The following projects are related to both calls: N. Title Coordinator 1 Reelin and GSK3 as therapeutic targets in Alzheimer's disease Ávila de Grado, Jesús 2 BESAD-P: Biomarkers of Early Stages of Alzheimer Disease- Prevention Ferrer Abizanda, Isidro 3 Glial activation during the neuroinflammatory process: a potential therapeutic target for Alzheimer's disease Vitorica Ferrández, Francisco Javier 4 Neuroprotection in Huntington's disease García de Yébenes Prous, Justo 5 Generating a dopaminergic neuronal model from induced pluripotent stem cells from patients with Parkinson's disease associated to mutations in the LRRK2 gene 6 Consortium to generate a common database aimed at implementing clinical and basic research on neuromuscular diseases 7 The transcription factor Nrf2 as a new therapeutic target for Parkinson's disease 8 Onset and progression of Parkinson's disease. Vulnerability of the nigrostriatal pathway, events at origin and destination 9 Generation of dopaminergic neurons from somatic cells of parkinsonian patients with cognitive failure 10 Multicenter study on LCR and neuroimaging biomarkers in the continuum preclinical- prodromal Alzheimer's disease (SIGNAL Study) Tolosa Sarró, Eduardo Illa Sendra, Isabel Cuadrado Pastor, Antonio Obeso Inchausti, José Ángel Moratalla Villalba, Rosario Lleó Bisa, Alberto Those ten already completed projects have actively involved a total of 49 CIBERNED research groups, in addition to some others external, representing 84.5% of the existing groups. The third call for CIBERNED cooperative projects was launched in May 2013, which was again externally evaluated by the ANEP and resolved in early September awarding 5 new collaborative projects, which are listed below: 261

268 N. Title Coordinator 11 Emergent properties of the neuron-glia relationship underlying neurodegeneration and dementia in Alzheimer's disease 12 Mitochondrial dynamics and mitophagy as therapeutic targets in Parkinson's and Huntington's diseases 13 Identification and molecular characterization of cannabinoid receptors subpopulations in poliglutaminopatías 14 Role of GSK-3β in the cortical circuits alterations occurring in Alzheimer's disease 15 Synaptic degeneration and dysregulation of adult neurogenesis in murine models of neurodegeneration Torres Alemán, Ignacio Soriano García, Eduardo Guzmán Pastor, Manuel Iglesias Vacas, Teresa Fernández Chacón, Rafael This third call for projects, funded with 688,000 per year for two years, involves 23 CIBERNED research groups and one external associated group. These projects were initiated during the last quarter of 2013 and have a duration of two years. A brief description of the scientific activity carried out by each of the projects in its first year is as follows: Project 11: Emerging properties of neuron-glia relationship underlying neurodegeneration and dementia in Alzheimer s disease Principal Investigator Torres Alemán, Ignacio Vicario Abejón, Carlos Vitorica Ferrández, Francisco Javier Comella Carnicé, Joan Xavier Gutiérrez Pérez, Antonia Moratalla Villaba, Rosario Boada Rovira, Mercé Institution CIBERNED, Cajal Institute CSIC, Madrid CIBERNED, Cajal Institute CSIC, Madrid CIBERNED, University of Seville Vall d'hebron University Hospital, Barcelona University of Málaga Associated group, CIBERNED, Cajal Institute CSIC, Madrid Associated group, Fundación ACE, Barcelona Summary In this collaborative project, we proposed as a general objective to determine changes associated with neuroinflammatory processes that appear throughout Alzheimer s disease (AD) in the neuron-glia relationship and its impact on the development of synapto-toxicity and subsequent neuronal death. The ultimate goal would be identifying new therapeutic targets to influence neuronal degeneration and therefore the clinical progression of AD. In particular we are studying the emergent properties of microglia, astroglia and neurons considering the functional relationship between them and considering the Aβ/tau binomial as the running thread triggering these new cellular properties. We are determining the impact of Aβ/tau in the process of 262

269 2014 Annual Report Cooperative Research microglial differentiation characteristic of AD as first adaptive event. In astrocytes we analyze the induction by Aβ, likely through microglial inflammatory mediators, of an interaction between the phosphatase calcineurin (involved in inflammatory processes), the transcription factor FOXO (involved in responses to oxidative stress) and adaptive changes in the neuroprotective factor IGF-I signaling. In neurons we analyze the interrelationship between survival mechanisms induced by the pro-inflammatory cytokine TNF and death mechanisms and inflammation where Fas-ligand is involved, as well as synapto-toxic processes involving for instance loss of synaptic plasticity and IGF-1 and BDNF synapto-trofic processes. Another part of the project seeks to develop disease cellular models to compare the findings in these models, including the necessary gathering of descriptive data in patient samples. We can summarize very briefly the results obtained so far in the following main findings: 1. Microglia suffers a degenerative process in the hippocampus of Alzheimer s patients (Braak stages V-VI). In the same samples, astroglia shows no sign of degeneration, in any case activation/ proliferation. Furthermore, both tau and extracellular Aβ are toxic to neurons and microglia but not to astrocytes, which seems to get activated. 2. The interaction between calcineurin and FOXO in astrocytes activated by inflammatory cytokines present in the AD brain is modulated by small molecule drugs (produced by the company associated with the consortium) which in turn inhibit the neuronal death induced by the inflammatory process. 3. Glial-derived pro-inflammatory cytokines (TNF-alpha) sensitize neural cells (neuroblastoma) to apoptotic death via Fas. In addition, Aβ inhibits the FAIM-L death receptors inhibitor in primary neurons, essential element of the protective effect of TNF on neurons. FAIM-L is reduced in AD brains and in mouse models of the disease. 4. Although we have not been able to work with human samples as planned (see below), we have characterized in detail and standardized the process of obtaining ipscs from human tissue from healthy controls and initiated a characterization of autophagy processes in those ipscs. Project 12: Mitochondrial dynamics and mitophagy as therapeutic targets in Parkinson s Disease and Huntington Principal Investigator Soriano García, Eduardo Vila Bover, Miquel Trullás Oliva, Ramón Alberch Vié, Jordi Tolosa Sarró, Eduardo Institution CIBERNED, University of Barcelona CIBERNED, Vall d'hebron University Hospital, Barcelona CIBERNED, IDIBAPS-CSIC, Barcelona CIBERNED, University of Barcelona CIBERNED, Clinic Hospital, Barcelona Summary Mitochondrial dysfunction is a key process in the pathogenesis of many neurodegenerative diseases, in particular Parkinson s disease (PD), and Huntington s disease (HD). In addition to mitochondrial disorders related to cellular respiration and production of ATP, calcium homeostasis and oxidative stress, recent studies suggest a second type of mitochondrial alterations that converge on two essential processes for 263

270 neuronal viability and physiology: mitochondrial transport and dynamics, and mitochondria recycling thorugh mitophagy. Although some of the genes involved in these processes are known, the exact mechanisms that regulate mitochondrial dynamics and mitophagy in neurons, as well as their involvement in the pathogenesis of PD and HD -especially in early stages- remain unknown. The objective of this project is to dissect the role of mitochondrial proteins recently identified by the consortium groups as regulators of mitochondrial dynamics and mitophagy processes (the Armcx/Armc10 family, Pentraxin1/ NP1 and HTT) and in the pathogenesis of PD and HD. Second, we will determine its potential therapeutic utility in treating these disorders. To this end, three specific objectives are proposed: 1. To dissect the molecular mechanisms by which the Armcx/Armc10, NP1 and HTT genes regulate mitochondrial dynamics and mitophagy. 2. To characterize the pathological alterations in mitochondrial dynamics and mitophagy in PD and HD (in brains of patients as in cellular models of disease, including fibroblasts and neurons differentiated from patient-derived ipscs) and their relationship to the above mentioned genes. 3. To investigate whether manipulating the expression of those genes is able to reverse the PD and HD associated mitochondrial deficits and restore function/neuronal viability both in vitro (fibroblasts and neurons from patient-derived ipscs) as in experimental models in vivo. The results of this project will help to decipher the role of the alterations of mitochondrial dynamics and mitophagy in the pathogenesis of PD and HD, and will be useful in assessing whether Armcx/Armc10, NP1 and HTT genes may represent new therapeutic targets for the treatment of these diseases. Project 13: Identification and molecular characterization of cannabinoid receptors subpopulations in polyglutaminopathies Principal Investigator Guzmán Pastor, Manuel Fernández Ruiz, Javier McCormick, Peter Joseph Mena Gómez, María Ángeles Institution CIBERNED, Complutense University of Madrid CIBERNED, Complutense University of Madrid CIBERNED, University of Barcelona CIBERNED, Ramón y Cajal University Hospital, Madrid Summary Understanding the process of neuronal survival is essential for the characterization of the etiology and progression of neurodegenerative diseases and, therefore, for the rational design of therapies for treatment. In this context, in recent years the four groups of this consortium have studied how the cannabinoid system modulates these processes, and what may be the therapeutic soundness of cannabinoid receptors as neuropharmacological targets. Specifically, the CB1 cannabinoid receptor is the most abundant mammalian brain metabotropic receptor and it is highly expressed in the basal ganglia and cerebellum, where it plays an essential role in controlling the motor activity and coordination, respectively. It has been reported that expression of the CB1 receptor decreases in basal ganglia of patients and animal models of Huntington s disease (HD), the most studied neurodegenerative disease among all caused by triplet expansion. Although there have been detailed studies on the protective role of cannabinoid receptors in HD models, we still do not know the precise subpopulations of cannabinoid receptors involved in the neuroprotective effects. The existence of subpopulations for the CB1 receptor have recently been suggested by the discovery that this receptor has different efficacy to signal depending on its location on GABAergic and glutamatergic terminals and the possibility to form heteromers with 264

271 2014 Annual Report Cooperative Research other metabotropic receptors in transfected cells or tissues ex vivo. Therefore, the overall objective of this project is to identify and molecularly characterize subpopulations of cannabinoid receptors (CB1 and CB1- adenosine receptors or CB1-other cannabinoid receptors heteromers) and their potential as new targets to induce neuroprotection in poly-glutaminopathies. These goals can be broken down into the following specific objectives: 1. Studying the differential expression of the CB1 receptor heteromers and its neuroprotective role in GABAergic and glutamatergic terminals. 2. Studying the neuroprotective role of the CB1 receptor and its heteromers in murine models of two poly-glutaminopathies (HD and SCA3) and in animals in which the selective activation of neuronal populations is induced by the pharmacogenetics technique Designer Receptors Exclusively Activated by Designer Drugs (DREADD). 3. Studying possible alterations in the expression of CB1 and its heteromers in postmortem brain tissue from patients with two poly-glutaminopathies (HD and SCA3). These studies, which will combine the complementary expertise and capabilities of the four groups of the consortium will help to clarify the molecular basis and potential clinical relevance of two important issues: (i) whether the alterations of the cannabinoid system are involved in the pathogenesis of polyglutamine disorders, and (ii) whether the stimulation of the cannabinoid signaling promotes neuroprotection and thus delay the onset and/or attenuate the progression of these diseases. Project 14: Role of GSK-3β in cortical circuits alterations occurring in Alzheimer s disease Principal Investigator Iglesias Vacas, Teresa Ávila de Grado, Jesús de Felipe Oroquieta, Javier Naranjo Orovio, José Ramón Lleó Bisa, Alberto Institution CIBERNED, Institute of Biomedical Research CSIC-UAM, Madrid CIBERNED, Center of Molecular Biology "Severo Ochoa" CSIC- UAM, Madrid CIBERNED, Cajal Institute CSIC, Center of Biomedical Technology, Madrid CIBERNED, National Center of Biotechnology CSIC, Madrid CIBERNED, Santa Creu i Sant Pau Hospital, Barcelona Summary It is generally considered that altered calcium levels and over-activation of glycogen synthase kinase-3β (GSK-3β) are directly related to the pathogenesis of AD, hence therapies aimed at modulating calcium homeostasis and/or inhibiting GSK-3β, applied in early stages of the disease, could have great potential for treatment. In this project we are analyzing modifications/alterations associated with GSK-3β hyperactivity, such as the levels and degree of phosphorylation of substrates like tau and Kidins220 in the continuum of AD and its relationship to early changes in the calcium neuronal homeostasis, in order to identify those changes associated with the disease in preclinical or prodromal stages that can help make an early diagnosis of AD. 265

272 For our studies will are mainly using murine models that inducibly overexpress GSK-3β as well as brain samples and cerebrospinal fluid (CSF) from patients at different stages of AD. With this work we are deepening our understanding of the molecular basis by which GSK-3β is involved in the etiopathogenesis of AD. Moreover, we are also studying possible markers that may facilitate early detection of the disease in order to be able to intervene therapeutically before major signs of cognitive impairment appear. Project 15: Synaptic degeneration and dysregulation of adult neurogenesis in mouse models of neurodegeneration Principal Investigator Fernández Chacón, Rafael Lucas Lozano, José Javier Fariñas Gómez, Isabel CIBERNED, University of Seville Institution CIBERNED, Center of Molecular Biology "Severo Ochoa" CSIC- UAM, Madrid CIBERNED, University of Valencia Summary It is widely accepted that neurodegenerative processes show, from the early stages of the disease, a synaptic-type dysfunction. However, synaptic alterations are more difficult to analyze experimentally that the neuronal vulnerability, contributing to the lack of information on how they can participate in neurodegenerative processes. On the other hand, recent evidence show that synaptic activity can regulate adult neurogenesis processes and this may contribute to the progression of certain symptoms in neurodegenerative processes and to the possible brain palliative response to these processes. The identification, in recent years, of molecules involved in the neurotransmitters release and related with various neurodegenerative diseases has opened the possibility to study the connection between the maintenance of synaptic function and disorders such as Parkinson s (PD) or Huntington s disease (HD). Therefore, this consortium aims at studying the relationship between synaptic activity and neurodegeneration through a strategy that consists in the use of mouse models in which three of these molecules are affected: CSP-alpha, alpha-synuclein and huntingtin in order to study 1) adult neurogenesis processes in the context of synaptic alterations and 2) endoplasmic reticulum stress as a basic pathogenic mechanism in response to synaptic alterations. This consortium will study the relationship between synaptic activity, regulation of adult neurogenesis and neurodegeneration in several genetically modified mouse models: a model of polyglutamine-mediated toxicity (R6/1mice with mutant huntingtin), a model of presynaptic degeneration [Cysteine String Proteinalpha (CSPalpha)-deficient mice] and models with genetic modifications of the synaptic protein alphasynuclein. Previous results suggest an intriguing connection, yet little explored, between huntingtin and CSP in neurodegenerative mechanisms. On the other hand, pathological interactions between CSP-alpha and alpha-synuclein have been described and both proteins regulate neurotransmitter release by acting on the SNARE complex. Unpublished data indicate an alteration of the hippocampal neurogenesis and susceptibility to epilepsy due in part to a deficit of GABAergic inhibition in CSP-alpha KO mice. In this project we will investigate if they have effects on neurogenesis by studying the neurogenic niches in the subgranular and subventricular zone. In particular, we will characterize the effects on neurogenesis of the degeneration of synaptic terminals that innervate that neurogenic niche and the synaptic dysfunction in the various models. In parallel, we will study the specific weight of the endoplasmic reticulum stress 266

273 2014 Annual Report Cooperative Research as a cause of neuronal death in our murine models and the possibility of reversing the degeneration therapeutically by pharmacological inhibition of the that mechanism. The projects from the third call will remain active during In parallel, during a forth call for proposals have been launched in 2014, with a budget significantly reduced compared with previous calls due to the need to cover the funding by CIBERNED international calls traditionally funded directly by the ISCIII (in particular those of JPND and ERA-NET). After the scientific evaluation of the ANEP, the call was resolved by awarding only two projects: Nº Title Coordinator 16 Onset and progression of Parkinson's disease: role of glial activation Rodríguez Oroz, María Cruz 17 Epigenetic mechanisms involved in the etiology and progression of rapidly progressive neurodegenerative dementias Calero Lara, Miguel This fourth call for proposals is funded with 532,074 for two years, and involves the active participation of eight CIBERNED research groups. A summary of the objectives set for the projects approved in the fourth call, initiated during the second half of 2013 is presented below. Project 16: Onset and progression of Parkinson s disease: role of glial activation Principal Investigator Rodríguez Oroz, María Cruz Matute Almau, Carlos Obeso Inchausti, José Ángel Rodríguez Díaz, Manuel Institution CIBERNED, Biodonostia Research Institute, San Sebastián CIBERNED, University of the Basque Country, Bilbao CIBERNED, Center for Applied Medical Research, University of Navarra, Pamplona CIBERNED, University La Laguna, Tenerife Summary Parkinson s disease (PD) is characterized by degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc) and the presence of cytoplasmic inclusions containing aggregates of alphasynuclein (a-syn). In most cases it is sporadic, but a genetic cause exist in 5-10% of the cases, with the most common mutations lying in LRRK2 gene. GWAS studies point to an important role of this gene in sporadic PD. The first motor signs in PD appear when dopamine loss is about 40-60%, suggesting that neurodegeneration precedes diagnosis by several years. In fact, asymptomatic carriers of LRRK2 mutations show a dopaminergic depletion years before the diagnosis of PD. Several mechanisms have been implicated in the pathogenesis and progression of PD, among which glial activation and neuroinflammation have become highly relevant. Studies in patients with advanced PD and in animal models of acute and severe dopaminergicv injury (6-OHDA rats and MPTP monkeys) show numerous evidence of an activation of microglia and astrocytes and high levels of proinflammatory cytokines (PIC) in the striatum and SNc associated with dopaminergic death. However, astrocytes may 267

274 also have a neuronal protective role through various mechanisms. Moreover, it has been hypothesized that LRRK2 mutations are associated with a proinflammatory state in microglial cells. Thus, existing data show a strong interaction between inflammation and dopaminergic death, although no human studies or animal models used so far allow to elucidate the role of microglia and astrocytes in the PD (onset/ progression or a consquence of dopaminergic death) nor is it known whether PD-causing mutations induce pro-inflammatory changes that can lead to dopaminergic degeneration. In this work, we intend to clarify the role that astrocytes and microglia have in PD through various complementary approaches. To that end, we will analyze the relationship between glial cells, neuroinflammation and dopaminergic neuronal death in relation to sporadic PD and LRRK2 mutations. Studies will be performed in cell cultures derived from patients with and without LRRK2 mutation, asymptomatic carriers of the mutation and in vivo studies in animal models of progressive nigrostriatal injury as well as in patients and asymptomatic carriers. In particular, we will generate and characterize astrocyte (Ast) differentiated from induced pluripotent cells (ips) derived from fibroblasts of patients with sporadic PD and LRRK2 mutations as well as asymptomatic carriers of these mutations. We will study in vivo the survival, proliferation, differentiation and migration of Ast-iPS implanted in the striatum of control rats and with dopaminergic denervation to know its role in the onset and progression of dopaminergic death. The temporal relationship between nigrostriatal lesion and microgrial activation will be defined in vivo by PET in the model overexpressing mutant human a-syn A53T in rat and by progressive MPTP treatment in monkeys. In addition, the relationship between striatal dopaminergic denervation and the presence of inflammatory changes by PET will be clinically validated in asymptomatic carriers of the LRRK-2 mutation and in eraly sporadic PD patients. General aim To study the relationship between glial cells, neuroinflammation and dopaminergic neuronal death in relation to sporadic PD and mutations in LRRK2 thorugh studies in cell cultures derived from patients with and without mutations, asymptomatic carriers of the mutation and in vivo studies in animal models of progressive dopaminergic lesion in patients. Specific aims 1. Generation and characterization of astrocytes (Ast) differentiated from induced pluripotent stem cells (ips), fibroblasts derived from patients with sporadic PD and with LRRK2 mutations and asymptomatic carriers of this. 2. To study in vivo survival, proliferation, differentiation and migration of astrocytes derived from ips (Ast-iPS) implanted in control rat striatum and after dopaminergic denervation to know its role in the onset and progression of nigrostriatal degeneration 3. To define in vivo the temporal relationship between nigrostriatal lesion and microgrial activation by PET ([11C]DTBZ and [11C]PK11195) through overexpression of A53T a-syn (rat) and progressive MPTP treatment (monkey). 4. To analyze the relationship between striatal dopaminergic denervation and the presence of inflammatory changes by PET ([18F]DOPA and ([11C]PK11195) in asymptomatic LRRK-2 mutation carriers and early sporadic PD patients. 268

275 2014 Annual Report Cooperative Research Project 17: Epigenetic mechanisms involved in the etiology and progression of rapidly progressive neurodegenerative dementias Principal Investigator Calero Lara, Miguel Ferrer Abizanda, Isidro del Río Fernández, José Antonio Sánchez Juan, Pascual Institution CIBERNED, Institute of Health Carlos III - CIEN Foundation, Madrid CIBERNED, Bellvitge Biomedical Research Institute, Barcelona CIBERNED, Catalonian Bioengineering Institute, Barcelona CIBERNED, Marqués de Valdecilla Foundation, Santander Summary Rapidly progressive neurodegenerative dementias (RPND) in the elderly include Creutzfeldt-Jakob disease (CJD), rapidly progressive dementia with Lewy bodies (DLBrp) and rapidly progressive Alzheimer s disease (RPAD), in which the cognitive impairment progresses rapidly and patients die within a period of a few months. The rapid evolution of the disease hinders its early detection and reduces the possibility of therapeutic interventions. Very little is known about the causes of the rapid development in the case of CJD and virtually nothing about the rapid forms of AD and DLB. Only a small proportion of RPND is associated with autosomal dominant point mutations in different genes (PRNP in CJD, APP, PSEN1 and PSEN2 in EA; SNCA, SCNB and GBA in DLB) while most of the cases are sporadic. Recent studies suggest a regulation of neurodegenerative diseases through epigenetic mechanisms. These mechanisms include methylation of CpG-rich regions especially in promoter regions of DNA, histone modifications that allow or block the accessibility of transcription factors to the binding sites on DNA and the involvement of various non-coding RNAs including micrornas that facilitate or inhibit translation of RNA into proteins. Moreover, the hypermutability of CpG dinucleotides may have a role in the etiology of various degenerative diseases. However, no study on CpG dinucleotide hypermutability in RPND exist. The generation of patient-derived induced pluripotent stem cells (ipsc) may represent an excellent model of human neurodegenerative diseases in which to study epigenetic alterations. This project focuses on the study of epigenetic mechanisms involved in the etiology and progression of RPND compared with their slower evolution counterparts and has the following main objectives: 1. Identify epigenetic molecular mechanisms that modulate the expression in brain tissue from RPND: CJD, RPAD, DLBrp. 2. Analyze whether CpG dinucleotide hypermutability in genes associated with CJD, AD and DLB plays a role as a regulatory mechanism in RPDN. 3. Identify epigenetic and transcriptomic signatures in differentiated ipsc. 4. Promote a sub-classification and definition of RPND endophenotypes according to neuropathological and molecular features based on epigenetic modulations. 5. Elucidate whether the observed changes may be useful as biomarkers in biological samples affordable in life such as blood and CSF. 269

276 CIBERNED cooperative program has so far invested an overall budget of about 5,422,000, distributed among the 17 cooperative projects mentioned above. A total of 55 CIBERNED research groups have actively participated in the cooperative program, which represents 94.5% of all groups belonging to CIBERNED during this period, along with a dozen of outside associated groups. In addition, 25 groups (43%) have participated in more than one cooperative project and projects are currently still active, with the involvement of 31 research groups CIBERNED (57%). A table is presented below with an estimate of the evolution of the investment made so far in the program of cooperative research as per project and participating group: Call Budget Duration in years Total number of groups Number of projects Average budget for project and year Average budget for project and group ,322, , , ,192, , , ,376, , , , , ,

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279 2014 Annual Report International Relations International scientific collaboration increases more and more, not only because of the availability of international funding and the drive of modern communication technologies, but also because science itself has become a truly international collaborative activity. In particular, the scope and scale of the problem of neurodegenerative diseases in today s society require a global response to confront this great challenge and thus has been recognized by various international institutions such as the European Union (EU), the Organization for Economic Cooperation and Development (OECD), the World Health Organization (WHO), etc.), and the industrialized countries that constitute the G8. The leaders of governments, businesses and academia also recognize the need for a coordinated strategy to address this major global challenge for health systems. There is consensus among all stakeholders on the need to build capacities, infrastructures and R&D resources in the field of neurodegenerative diseases. There is also a pressing need for global participation and a commitment to a significant increase in investment in skills and resources to reduce the duration of these chronic brain pathologies and/or the number of people at risk. This budgetary effort should be accompanied by sound policies and legislative initiatives to encourage public-private partnerships. History has shown that collaboration between academic researchers, government agencies and pharmaceutical and biotechnology companies is an essential ingredient in promoting this type of ambitious initiatives, especially when resources are limited. In this context, in recent years CIBERNED, has given a boost to its relations with international organizations in the area of research in neurodegenerative diseases such as the EU Joint Programme for Research in Neurodegenerative Diseases (JPND) and the Network of Centres of Excellence in Neurodegeneration (CoEN), among other initiatives. European Union Joint Programming on Neurodegenerative Disease Research (Jpnd) The EU Joint Programming for Research in Neurodegenerative Diseases (JPND) is an innovative collaborative research initiative created to address the growing challenges posed by these disorders. The JPND is a pioneering example of joint programming for the promotion of research within the European Union aimed at scientific challenges requiring a response that exceeds the capacity of a single country, based on the alignment of national research programs devoted to these challenges. Its objective is to enhance the impact of research by aligning existing national research programs and the identification of common objectives whose scope would benefit from joint action. The JPND Scientific Advisory Committee has significant participation of two CIBERNED researchers, Drs. Jesús Avila and Jesús de Pedro, as well as Dr. Angel Cedazo-Mínguez, from the Karolinska Institute in Stockholm and member of the CIBERNED Scientific External Advisory Committee. The Research Strategy designed by JPND provides a framework for future investments and shows that the research effort within the European Union can be leveraged to improve care on prevention, diagnosis and treatment of patients suffering from these diseases. To achieve impact there is a need to encourage novel as well as multidisciplinary approaches, and to strengthen and extend existing capabilities across the full spectrum of basic, clinical, health and social care, and translational research. To that end, a number of priority areas for future research have been identified: The origins of neurodegenerative diseases; Disease mechanisms and models; Disease definition and diagnosis; Treatment and prevention; Health and social care. This Research Strategy also provides a framework of opportunities for countries involved in JPND and willing to participate in joint actions, which will be implemented through co-operative activities that realign or link national investments to achieve increased impact, and the provision of new funding. A guiding principle for its delivery will be that the research to be supported is of the highest scientific quality. In this regard, during 2011 took place the first call for European research projects JPND. Under the theme Optimization of biomarkers and harmonization of their use in the clinic, a total of four transnational projects were awarded for the period As described below, two of the approved projects have participation CIBERNED research groups. 273

280 BIOMARKAPD: Biomarkers for Alzheimer s disease and Parkinson s disease Total Funding 8.5 M (approx.) Start date June, 2012 Duration Coordinator CIBERNED participation 3 years Bengt Winblad, Karolinska Institutet, Stockholm, Sweden Alberto Lleó and Javier Sáez-Valero groups Project partners Judes Poirier, McGill University, Quebec, Canada Bengt Winblad, Karolinska Institutet, Stockholm, Sweden Lennart Minthon, Skåne University Hospital, Sweden Henrik Zetterberg, The Sahlgrenska Academy, University of Gothenburg, Sweden Gunhild Waldemar, Rigshospitalet, Copenhagen Univ Hospital, Denmark Niels Heegard, Statens Serum Institut, Denmark Hilkka Soininen, University of Eastern Finland, Finland Juha Rinne, Turku University Hospital, Finland Bruno Dubois, Salpetriere University Hospital (UPMC - University Paris 6), France Sylvain Lehmann, CHRU de Montpellier, France Armand Perret-Liaudet, Université Lyon 1, France Jens Wiltfang, Universität Duisburg-Essen, Germany Thomas Klockgether, German Center for Neurodegenerative Diseases (DZNE), Germany Brit Mollenhauer, Paracelsus-Elena-Klinik + Georg August University Göttingen, Germany Piotr Lewczuk, Universitätsklinikum Erlangen, Germany Markus Otto, University of Ulm, Germany Katrin Marcus, Ruhr University Bochum, Germany Dieter Willbold, Heinrich-Heine University Medical School Duesseldorf, Germany Lutz Froehlich, Central Institute of Mental Health Mannheim, Germany Thomas Arendt, University of Leipzig, Germany Magda Tsolaki, Aristotle University of Thessaloniki, Greece Elisabeth Kapaki, University of Athens, Greece Brian Lawlor, Trinity College Dublin, Ireland Andrea Urbani, IRCCS Foundation Santa Lucia, Italy Fabrizio Tagliavini, IRCCS Foundation Carlo Besta, Italy Giovanni Frisoni, IRCCS Foundation San Giovanni Di Dio Fatebenefratelli, Italy Paul Wilmes, University of Luxembourg, Luxembourg Pieter Jelle Visser, String of Pearls Initiative Neurodegeneration, The Netherlands Marcel Verbeek, Radboud University Medical Centre, Nijmegen, The Netherlands Tormod Fladby, Akershus University Hospital, Norway Jacek Kuznicki, International Institute of Molecular and Cell Biology (IIMCB), Poland 274

281 2014 Annual Report International Relations Andrzej Szczudlik, Jagiellonian University College of Medicine (UJ), Poland Tomasz Gabryelewicz, IMDiK, Mossakowski Medical Research Centre, Poland Marzena Zboch, Scinawa, Wroclaw Medical University, Poland Alexandre Mendonca, Instituto de Medicina Molecular, Portugal Catarina Resende de Oliveira, University of Coimbra, Portugal Odete da Cruz e Silva, University of Aveiro, Portugal Norbert Zilka, Slovak Academy of Science, Slovakia Uros Rot, University Medical Centre, Ljubljana, Slovenia José Luis Molinuevo, Hospital Clinic, Barcelona, Spain Alberto Lleó, Santa Creu i Sant Pau Hospital, Barcelona, Spain Joan Gispert, Pasqual Maragall Foundation, Barcelona, Spain Javier Sáez Valero, University Miguel Hernández-CSIC, CIBERNED, Spain Christoph Hock, University of Zurich, Psychiatric & University Hospital, Switzerland Panteleimon Giannakopoulos, University of Geneva Hospital, Switzerland Sermin Genc, Dokuz Eylul University, Turkey Esen Saka Topcuoglu, Hacettepe University, Turkey Hakan Gurvit, Istanbul University, Turkey Alzheimer s and Parkinson s diseases (AD and PD) are the two most common neurodegenerative conditions. They cause major costs for society and suffering and death for millions of patients around the globe. In Europe, more than 8 million individuals have AD or PD. Current treatments are symptomatic but do not stop the underlying disease process. Using biomarkers, we can detect biochemical changes that show when neurons start to die. There are also biochemical tests for brain changes that are specific to AD and PD. Studies suggest that such abnormalities start to appear years before onset of symptoms. If we want to do something substantial about these diseases, we need to diagnose them early, before too many neurons have been lost, and then treat them with drugs that inhibit the destructive process. Such drugs are in development. However, in these very early stages of the disease we cannot rely on clinical symptoms, as they may be very subtle, or even absent. Instead, research tells us that we could use biomarkers for disease-specific pathologies. Established biomarkers exist for early AD and promising candidates are underway for PD. However, a major problem today is the lack of standardisation regarding exactly how to perform and use the biomarker tests. In BIOMARKAPD we detail how we will standardise the biomarker measurements across Europe, how to collect samples, how to perform the measurements and how to interpret the results. We will also create a biobank with samples from well characterised AD and PD patients, including patients in very early disease stages, as well as neurologically healthy controls. These samples will be used to develop new and better assays and to test new and better biomarker candidates. Finally, we will develop certified reference materials that can be used to harmonise assays that are used to measure the different biomarkers. The deliverables of the proposal will have a major influence on clinical research and drug development for neurodegenerative conditions in general and for AD and PD in particular. They will impact these types of efforts globally and make Europe world-leading in this area. 275

282 DEMTEST: Biomarker based diagnosis of rapid progressive dementiasoptimisation of diagnostic protocols Total Funding 2.2 M (approx.) Start date May, 2012 Duration Coordinator CIBERNED participation 3 years Inga Zerr, University Medical Center Göttingen, Germany Pascual Sánchez-Juan (Berciano group) and Miguel Calero (de Pedro group) Project partners Inga Zerr, University Medical Center Göttingen, Germany Carsten Korth, Heinrich Heine University Medical School, Düsseldorf, Germany Hans Kretzschmar, Ludwig-Maximilians-University, Munich, Germany Jean-Louis Laplanche, Hopital Lariboisiere AP-HP, France Olivier Andreoletti, UMR-INRA-EVNT, France Theodoros Sklaviadis, Aristotle University of Thessaloniki, Greece Stefano Ruggieri & Maurizio Pocchiari & Anna Ladogana, University Sapienza, Rome, Italy Pawel Liberski, Medical University of Lodz, Poland Catarina Resende Oliveira, University of Coimbra, Portugal Eva Mitrová, Slovak Medical University Bratislava, Slovakia Gorazd Bernard Stokin, University Psychiatric Hospital, Ljubljana, Slovenia Miguel Calero, National Institute of Health Carlos III, Spain Pascual Sanchez-Juan, IFIMAV and CIBERNED, Marqués de Valdecilla University Hospital, Spain Anna-Lena Hammarin, Swedish Institute of Communicable Disease Control, Sweden Adriano Aguzzi & Herbert Budka, University Hospital Zürich, Switzerland John Collinge, University College London, United Kingdom Robert G. Will, Western General Hospital, United Kingdom DEMTEST has established a large European and global collaboration between national surveillance units and dementia research centres, facilitating cooperation between neurologists, neuropathologists and neuroscientists. Our goals are: To harmonize the protocols involved in patient documentation, biomaterial sampling/storage, biomarker testing/assay analysis and data sharing. To standardise a more precise diagnosis in patients with rapidly progressive dementia by analysis of the biochemical markers in the cerebrospinal fluid and blood. To improve CSF based diagnosis in dementia by application of new methodologies. We work on standardisation of tests that are currently available and harmonise their use between centers worldwide. We define standards for biochemically based diagnosis in most relevant rapid progressive dementia such as CJD and rapidly progressive Alzheimer s disease. We will improve innovative methods for amplification assays for mis-folded proteins and introduce their use into clinical routine. 276

283 2014 Annual Report International Relations As an add-on value, we will define criteria for early differential diagnosis between rapidly progressive neurodegenerative or potentially reversible dementia. The second JPND call for collaborative research projects was launched in 2013 and was focused on two identified topics of particular interest such as i) risk factors (genetic, epigenetic, environmental) for developing neurodegenerative diseases such as Alzheimer s, and ii) medical and social care strategies for people living with these mitigating diseases. This call for projects was funded with a total of 29 million euros to be distributed between both topics: 19 millions for topic 1 and 10 millions for topic 2. A total of five transnational projects were awarded for the period , one of which has participation from CIBERNED: COURAGE-PD: Comprehensive Unbiased Risk factor Assessment for Genetics and Environment in Parkinson s Disease Total Funding 4.5 M (approx.) Start date January, 2014 Duration Coordinator CIBERNED participation 3 years Thomas Gasser, University of Tübingen, Germany Eduardo Tolosa group Project partners Thomas Gasser & Rejko Krüger University of Tübingen, Germany Alexis Elbaz, INSERM, France Stefano Goldwurm, Istituti Clinici di Perfezionamento, Italy Mathias Toft, Oslo University Hospital, Norway Nicholas Wood, University College London, United Kingdom Avi Orr-Urtreger, Tel Aviv Sourasky Medical Center, Israel Joaquim Ferreira, Institute of Molecular Medicine, Portugal Eduardo Tolosa, University of Barcelona, Spain Rudi Balling, University of Luxembourg, Luxembourg Peter Heutink, VU University Medical Centre, Amsterdam, The Netherlands Despite years of research and the considerable progress that has been achieved by identifying a number of genes and genetic risk factors for Parkinson s disease (PD), the cause for vast majority of cases is still unknown. It is suspected that a multitude of interacting genetic and environmental risk factors, rather than single genetic mutations or single toxins, are responsible for the disease in most instances. The project COmprehensive Unbiased Risk factor Assessment for Genetics and Environment in Parkinson s Disease (COURAGE-PD) takes an integrated approach to unravel these underlying factors. The genetic make-up of large, well-characterized patient cohorts from different ethnic populations will be deciphered. At the same time, environmental exposures will be documented, and interactions between both classes of risk factors will be elucidated using novel statistical methods. Finally, the relevance and validity of the findings will be tested in innovative cellular models, including neuronal cells, which will be generated from patient-derived induced pluripotent stem cells. We expect that this combination of state-of-the-art genetic technologies with a detailed ascertainment of 277

284 environmental modifiers will provide important clues to decipher the complexity of neurodegeneration in Parkinson s Disease. Centers of Excellence in Neurodegeneration (Coen) A major obstacle to the advancement of research on neurodegenerative diseases is the relative lack of common standards and mechanisms for validation of potentially relevant results in preclinical studies, and clinical studies based on population. One approach to deal with these challenges on a large scale is through more effective use of large centers and institutes, where there is already the necessary critical mass of resources and expertise. Increased collaboration between national centers of excellence should also provide the opportunity to accelerate progress in understanding the basic mechanisms of disease, and the identification of new therapeutic approaches. To this end, on June 10, 2010, the Canadian Institutes of Health Research (CIHR), the German Centre for Neurodegenerative Diseases (DZNE, Germany) and the Medical Research Council (MRC, UK) launched a funding initiative to establish a collaborative approach to research in neurodegenerative diseases, called Centers of Excellence in Neurodegeneration (CoEN). These founding members were later joined by other European institutions and thus, in December 2011 the CoEN membership application by CIBERNED- CIEN Foundation was approved, recognizing the scientific excellence in both basic and clinical science of the institution which became part of the CoEN Oversight Group. Current CoEN members are: Canadian Institutes of Health Research (CIHR), Canada Deutsche Zentrum für Neurodegenerative Erkrankungen (DZNE), Germany Medical Research Council (MRC), United Kingdom Flanders Institute of Biotechnology (VIB Flanders), Belgium Health Research Board (HRB) / Science Foundation Ireland (SFI), Ireland Ministero della Salute (MDS), Italy Centre of Excellence for Brain Research, Slovakia CIBERNED- CIEN Foundation, Spain The overlapping of the CoEN group members with those of the JPND will ensure that their complementary objectives progress in close cooperation with each other. This is accomplished through a two-step process, involving expert workshops for the analysis of needs, followed by a call for proposals for collaborative teams between PIs within the participating national Centers of Excellence. Since 2012, CIBERNED and CIEN Foundation form part of the Oversight Group to actively participate in the design of CoEN future scientific strategy. Both institutions are represented in the CoEN Oversight Group by Dr. Miguel Medina, CIBERNED Deputy Scientific Director and member of the Scientific Advisory Committee of the CIEN Foundation. The first phase of the CoEN initiative began in late 2010 and has been aimed at the development of common resources and methodological approaches that underpin future studies. Some of the key issues addressed are: the development and validation of cell and animal models of disease; development of new measures to define subgroups of patients for clinical studies; identification of new biomarkers to support translational research; the development and harmonization of the battery of cognitive tests for the diagnosis and monitoring of disease progression; and the establishment of common platforms for improved data analysis and exchange. Phase II of the initiative was launched during the year 2013, with the aim of catalyzing collaborative research between centers with a critical mass of resources and expertise to thus promote a radical change in research on neurodegeneration. To do this, the countries participating in CoENs contributed 278

285 2014 Annual Report International Relations a total amount of 5.5 million euros (of which Spain has provided 600,000 euros) in this call to establish an innovative and progressive research program to address the major challenges in this field. The call is intended to encourage the community to think outside the pre-established frameworks and stimulate new and creative approaches and solutions to the challenges of research in neurodegeneration. This call of Pathfinder projects intends to combine the strengths of research groups through Centers of Excellence in at least two partner countries to provide a truly collaborative effort and value that will advance our approach to research neurodegeneration. The projects would address issues that are not easily financed through standard grant mechanisms from CoEN partners, and is expected to further collaboration between Centers of Excellence, the projects would also serve to provide a platform for future collaboration with industry. A total of eighteen project applications were submitted, eight of which included a CIBERNED research group (as coordinator in four of them). The result of the evaluation of these applications by an international panel of experts resulted in the approval of five research projects, three of which inlude Spanish participation (which is an excellent 37.5% success rate for CIBERNED applications) and a total committed funding for two years of 570,000 euros. The three projects involving CIBERNED are described below: WNT signaling: biomarker and target evaluation in Alzheimer s disease Project partners: Antonio Cuadrado (coordinator, CIBERNED, Spain), James Woodgett (Canada) and Simon Lovestone (UK) The failure of biopharmaceutical pipelines to modify Alzheimer s disease (AD) progression indicates the need for new creative approaches. This project was established to determine if novel elements of the canonical and non-canonical WNT signaling pathways could be targeted to modify and monitor AD progression. During the first year of the CoEN-pathfinder grant, we have been using multidisciplinary approaches, including brain oriented drug screening, a novel proprietary mouse model of AD, new genetic and pharmacological tools for modulation of WNT/GSK3 and WNT/JNK pathways, and human blood samples from AddNeuroMed, DCF and EMIF platforms to identify novel biomarkers of AD progression. The end goal is to explore the impact on AD of novel WNT elements including but not limited to clusterin, DKK1 and NRF2. In vivo neuronal cell reprogramming for a new regenerative approach in Parkinson s disease Project partners: Vania Broccoli (coordinator, Italy), Alexander Dityatev (Germany) and José Luis Lanciego (CIBERNED, Spain) Neurodegenerative diseases cause a significant burden on the elderly population in Europe. Parkinson s disease (PD) affects 1.2 million people in Europe and with the increasing life expectancy this number will rise, putting more pressure on health care. Treatment of PD is only symptomatic, and therefore, there is an urgent need for more efficient therapies. Degeneration of mesencephalic DA neurons triggers the initial phases of PD, which raised the concept that cell replacement might represent a long-term restorative option for this neuropathology. Indeed, previous studies in PD patients have indicated that cell therapy has the potential to significantly sustain an enduring symptomatic relief. However, these studies suffered from lacking an ideal source of transplantable human DA neurons. We have recently developed a methodology that promotes transdifferentiation of mouse and human fibroblasts into functional induced dopaminergic neurons (idans), which display sophisticated neuronal properties including pacemaking firing activity, synaptic integration, and activity-dependent dopamine release. Therefore, idan cells offer an unprecedented cellular source with ideal features for cell therapy in PD, since they can be generated from the patients in high amounts. Here, we propose to push forward this technology by elaborating methods for direct in vivo reprogramming to further induce in situ local neuronal transdifferentiation in relevant animal models of PD (mouse and monkey). MicroRNA as novel therapeutic targets and disease biomarkers in Alzheimer s Disease, Frontotemporal dementia and Amyotrophic lateral sclerosis (NEURO-MIR) Project partners: Jochen Prehn (coordinator, Ireland), Andre Fischer (Germany), Pierre Lau (Flanders, Belgium), José J. Lucas (CIBERNED, Spain) 279

286 NEURO-MIR focuses on the role of microrna (mirna) in neurodegeneration. mirnas are small (~22 nucleotide) non-coding RNA molecules that control the expression of multiple genes. Previous data from the members of NEURO-MIR consortium have shown that mirna deregulation contributes to the pathogenesis of Alzheimer s disease and motor neuron disorders. NEURO-MIR aims to explore the full potential of mirnas as major contributors to disease progression and as therapeutic targets and biomarkers. NEURO-MIR also recognizes that the complexity of the biology of mirna can only be addressed through bioinformatics and computational modeling methods, so that it aims to develop a computational platform to address the role of mirnas in neurodegeneration at the systems, holistic level. International Congress for Research and Innovation in Neurodegenerative Diseases (Ciiien) During 23 rd and 24 th of September, 2014 took place in Barcelona the second International Congress on Research and Innovation in Neurodegenerative Diseases (CIIIEN), promoted by the Queen Sofia Foundation in collaboration with CIEN Foundation. The main objective of CIIIEN is having a forum in which to share progress and information of interest on neurodegenerative diseases among the scientific community. CIIIEN consolidates the merger of the two major scientific conferences neurodegenerative diseases in general and Alzheimer s disease in particular, organized in Spain: the X International Symposium Advances in Alzheimer s Disease, promoted annually by the Queen Sofia Foundation and CIEN Foundation, and the 8th CIBERNED Scientific Forum, that brings together every year more than 500 scientists constituting the CIBER on Neurodegenerative Diseases. Unifying both congresses is a first step in creating a new operating structure in the two main institutions devoted to research on neurological and neurodegenerative diseases in Spain: CIEN Foundation and CIBERNED, both under the Ministry of Economy and Competitiveness through the Institute of Health Carlos III. This new structure seeks greater effectiveness and efficiency in research, promoting the interaction of the different research groups. This second edition of CIIIEN was chaired by Her Majesty Queen Sofia and the scientific program consisted of an opening session and eight scientific sessions covering various aspects of cutting edge research in neurodegenrativas diseases, such as stem cells, biomarkers, preclinical changes, aging, immunity, genetics, mechanisms of neurodegeneration and neuroprotection. Among the speakers at the conference can be highlighted some international researchers who are leaders in their field of research such as Michael Heneka (University of Bonn), Vincenzo Bonifati (Erasmus University, Rotterdam), Ammar Al-Chalabi (King s College Institute of Psychiatry, London), Anders Fjell (University of Oslo), and Leslie M. Thompson (University of California, Irvine). Thus, this event establishes in its second edition as a meeting point for the world s leading experts in neurodegenerative diseases, enabling sharing of knowledge, working methods, new advances and discoveries in a field in which international cooperation and between different institutions is becoming increasingly important to obtain optimum results in research. 280

287 2014 Annual Report International Relations Bilateral Uk-Spain Meeting On March 18 took place in the Ernest Lluch Hall of the Institute of Health Carlos III a bilateral conference United Kingdom-Spain, organized by the British Embassy in Madrid and in collaboration with the Institute of Health Carlos III and the Sanitas Foundation, entitled Dementia: A Global Challenge. The aim of the conference was to bring together scientists, health professionals, policy advisers, companies and associations of health care services, to debate the challenge posed by dementia, an issue of national importance both for the UK and Spain. During the meeting was held a round table on the importance of research in dementia policies, with the participation of Robin Buckle, Director of Science Programs at the Medical Research Council UK (MRC); Jesús Ávila, Scientific Director of CIBERNED; Agustin Ruiz, Research Director and CSO of the ACE Foundation; Steve Parr, Director of Business Development in Ixico; Pablo Martínez-Lage, Head of Neurology at the CITA Foundation and member of CIBERNED group led by Dr. Adolfo Lopez de Munain; Martin Orrell, Professor at University College London; and Antonio Paez, Head of the Department of Clinical Operations at Grifols Institute. Among those attending were also Antonio Andreu, Director General of the Institute of Health Carlos III; Simon Manley, British Ambassador in Spain; Mercedes Vinuesa, Director General of Public Health; Lorraine Jackson, Director of Policy Assistance for Dementia of the UK Department of Health; and Joaquín Casariego, Director of Development and International Relations, Faculty of Medicine at the University Francisco de Vitoria. The conference constituted a meeting point for contrasting experiences and establish an excellent forum for discussion to reduce the personal, social and economic impacts posed by dementia. 281

288 I Aviesan-Alinnsa Symposium On February 6 took place in the Gustavo Pittaluga Aula Magna of the National Health School of the Institute of Health Carlos III, the First Symposium between the French Alliance for Life Sciences and Health (AVIESAN) and the Spanish Alliance for Research and Innovation in Health (ALINNSA). The aim of the initiative was to foster cooperation of existing agents, through the coordination, joint programming, internationalization and promotion of public-private partnerships. The conference was inaugurated by the Secretary of State for Research and Innovation of the Ministry of Economy and Competitiveness, Ms. Carmen Vela, and the Ambassador of France Mr. Jerome Bonnafont. In addition, André Syrota, President of AVIESAN, and Antonio Andreu, President of ALINNSA presented the alliance between the two initiatives. During the event, a joint working group (ALINNSA-AVIESAN Joint Meeting: Opportunities for future cooperation) was established, where experts from both countries shared, in several subgroups, knowledge on thematic priorities from a multidisciplinary perspective, identifying actions and opportunities for joint future collaboration: In the working subgroup on Developmental Neurobiology and Neurodegenerative Diseases, Miguel Medina, CIBERNED Deputy Scientific Director, presented the structure and scientific objectives of CIBERNED and CIEN Foundation, as well as some opportunities for potential future scientific collaborations. In this group also participated Miguel Calero, member of the CIBERNED group led by Jesús de Pedro. 282

289 Scientific Productivity and Other Activities SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PR OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIV PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY VITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIF AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIV PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY VITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIF AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIV PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY VITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIF AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES SCIENTIFIC PRODUCTIVITY AND OTHER ACT

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291 2014 Annual Report Scientific Productivity and Other Activities Analysis of Ciberned Scientific Productivity in 2014 To complete this section we provide a table of bibliometric indicators for 2013 and A quick analysis allows to observe a strengthened of the improved scientific production in 2014 (in line with the ongoing historical improvement observed in the last years since the creation of the Centre), not only in quantity but particularly in quality (percentage of publications in quartiles 1 and 2, number of publications in high impact journals, average impact factor, international collaborations). INDICATOR % CHANGE (A) Total number of publications (B) Number of publications in quartiles (Q) 1 y (C) ) Number of publications in Q1 y Q2 as percentage of total (A) (E) Number of publications in 1 st decile (F) Number of publications (1 st decile) as percentage of total (A) (G) Number of publications in Q1 (it include 1 st decile) (H) Number of publications in Q1 as percentage of total (A) (I) Number of publications in Q1 as percentage of total Q1 and Q2 (B) (J) Number of publications in Q (K) Number of publications in Q2 as percentage of total (A) (L) Number of publications (Q1 + Q2) led by CIBERNED groups (1st author or corresponding author) (M) Number of publications in Q1 and Q2 led by CIBERNED groups as percentage of total (B) (N) Number of publications (Q1 and Q2) not led by CIBERNED groups (1 st author or corresponding author) (O) Number of publications in Q1 and Q2 not led by CIBERNED groups as percentage of total (B) (P) Number of publications in Q1 and Q2 in which CIBERNED appears in the afiliation (as percentage of total (B)) (Q) Number of publications in Q1 and Q2 led by CIBERNED groups in which CIBERNED appears in the afiliation (as percentage of total (L)) (R) Number of publications in Q1 and Q2 not led by CIBERNED groups in which CIBERNED appears in the afiliation (as percentage of total (N)) (S) Number of publications in Q1 and Q2 with international groups

292 INDICATOR % CHANGE (T) Number of publications in Q1 and Q2 with international groups (S) as percentage of total number of publications (B) (U) Number of publications in Q1 and Q2 with international groups, and led by CIBERNED groups (V) Number of publications in Q1 and Q2 with international groups, and led by CIBERNED groups as percentage of all international publications (S) (W) Number of publications in Q1 and Q2 with other CIBERs (X) Percentage of publications in Q1 and Q2 with other CIBERs led by CIBERNED groups (Y) Number Percentage of published in Q1 y Q (Z) Percentage of Percentage of led by CIBERNED groups (A2) Addition of impact factors of journals (Q1 y Q2) in which articles have been published (B2) Average of impact factors of journals (Q1 y Q2) in which articles have been published (C2) Number of publications in journals with impact factor >15 Number of publications in Q1 and Q2 with 2 CIBERNED groups Number of publications in Q1 and Q2 with 3 CIBERNED groups Number of publications in Q1 and Q2 with 4 CIBERNED groups Number of publications in Q1 and Q2 with 5 CIBERNED groups Number of publications in Q1 and Q2 with 6 CIBERNED groups Cooperative projects Program Cooperative projects Program Cooperative projects Program Program 1 cooperative projects Program 2 cooperative projects Program 3 cooperative projects Projects with other CIBERs/RETICs/CIEN networks Basic-type projects Translational-type projects Number of publications indexed in ISI (Web of Knowledge). 286

293 2014 Annual Report Scientific Productivity and Other Activities Below are some figures summarizing some of the main bibliometric indicators for CIBERNED scientific output during 2014 and its evolution since 2008: Publications Ciberned 2014 D1 28% D1 28% Q1 51% Q1 51% Q2 21% Q2 21% Ciberned Yearly Output Evolution

294 Ciberned Output Evolution by Quartile/Decile N. publications Q1+Q2 1 st DECILE N. publications Q1+Q2 1 st DECILE Ciberned Excellence Publications (1 St Decile) Rate

295 2014 Annual Report Scientific Productivity and Other Activities Ciberned Number of Publications in 1 St and 2 Nd Quartile/1 St Decile D1 Q1 Q2 D1 Q1 Q Impact Factor Evolution Accumulated IF Average Accumulated IF IF 3500 Average IF