2016 Annual Report MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD

Transcription

1 2016 Annual Report MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD

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3 2016 Annual Report

4 2017 CIBERNED Centro de Investigación Biomédica en Red, Enfermedades Neurodegenerativas C/ Valderrebollo, Madrid Coordination and management of content: Miguel Medina Padilla José de Arriba-Enríquez Aina Frontera Sánchez Almudena Flores Junquera Soledad Valero Rodríguez Design and Editorial coordination: Duomo Comunicación

5 Index Letter from the Scientific Director... 7 Overview... 8 Aims Directory of research groups and associated institutions Geographic distribution of CIBERNED research groups Organizational structure Organization chart External scientific advisory committee Consortium members Scientific report Research programs Program 1: Alzheimer s disease and other degenerative dementias Program 2: Parkinson s disease and other neurodegenerative movement disorders Cooperative research International relations Scientific productivity and other activities Financial report Principal investigator index

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7 Letter from the Scientific Director One more year, the 53 groups that comprise our Network Center have carried out a fruitful work, which allows us to continue at the top of the Spanish Centers with publications of greater impact factor in the field of Biomedicine. This work is greatly appreciated, as well as that of the Steering Committee and the Management Office, and that of the Carlos III Institute of Health, which has maintained its financial support. This year more than 30 articles have been published in scientific journals with an impact factor (IF) greater than 10. Seven of these publications were in journals that exceed an IF of 23, such as Science, Cell or Lancet Neurology. In addition, given the translational research nature of our Center, new patents have been applied for, over 60 clinical trials have been actively maintained and some new clinical guidelines have been published. Various courses, scientific conferences and seminars have also been organized, and the Training Program has continue to be developed. In addition, I would like to congratulate those colleagues who have received international or national awards or have been awarded relevant European and international competitive projects. Finally, I would like to underline the high and valuable participation in our Scientific and Social Forums. Jesús Ávila de Grado Scientific Director In short, this has been a good year. Thank you all CIBERNED Annual Report / 7

8 Overview The Network 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 created with the mission of fighting 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. Since 2007 the CIEN Foundation has its headquarters in the Alzheimer Center of the Reina Sofia Foundation, created with the collaboration of CIBERNED and the CIEN Foundation, and located in Vallecas (Madrid). CIBERNED was founded under the auspices of the Carlos III Institute of Health, 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 multi-institutional 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 CIBERNED Annual Report / 8

9 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. Its own statutes govern CIBERNED and since the end of 2014 its activity is structured around two Scientific Programs: Program 1: Alzheimer s disease and other degenerative dementias. Program 2: Parkinson s disease and other neurodegenerative motor disorders. During 2016 CIBERNED has consisted of 53 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: researcher members physically work in their parent institutions they belong to while simultaneously and actively participating on CIBERNED s own cooperative research agenda. Thus, it is the result of a collaborative partnership between different institutions and the sum of 53 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. Depending on budget availability, it may also consider adding some new research group currently outside CIBERNED that satisfy the requirements of scientific quality and translational activity and aligns 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 Foundation of CIBERNED 2 Research programs 53 Research groups 2016 CIBERNED Annual Report / 9

10 Aims CIBERNED has as its ultimate goal the promotion of scientific and technical research of excellence in the field of human health, with the generic purpose of producing results quickly transferable to clinical practice in order to improve the health and wellbeing of patients suffering of neurodegenerative diseases, as well as their families and caregivers, providing in turn economic and social benefits. Among its specific aims, 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 therapeutic or/and preventive interventions. 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...) To involve society in the enormous medical and socioeconomic impact of neurodegenerative diseases and facilitate their involvement in the fight against neurodegeneration CIBERNED Annual Report / 10

11 Directory of research groups and associated institutions Group Principal investigator Institution Alberch Vié, Jordi Ávila de Grado, Jesús Bullido Gómez-Heras, Mª Jesús Camins Espuny, Antonio Canela Campos, Enric Isidre Cantero Lorente, José Luis Carro Díaz, Eva Ceña Callejo, Valentín Comella Carnicé, Joan Xavier Cuadrado Pastor, Antonio de Felipe Oroquieta, Javier de Pedro Cuesta, Jesús del Río Fernández, José Antonio Fariñas Gómez, Isabel Fernández Chacón, Rafael Fernández Ruiz, Javier Ferrer Abizanda, Isidro Fuentes Rodríguez, José Manuel García Verdugo, José Manuel González Castaño, José Gutiérrez Pérez, Antonia Guzmán Pastor, Manuel Iglesias Vacas, Teresa Infante Ceberio, Jon Kulisevsky Bojarski, Jaime Labandeira García, José Luis Lanciego Pérez, José Luis Lleó Bisa, Alberto López Barneo, José López de Ceballos Lafarga, María López de Munain Arregui, Adolfo Lucas Lozano, José Javier Matute Almau, Carlos Mengod Los Arcos, Guadalupe Moratalla Villalba, Rosario University of Barcelona Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid Autonomous University, Madrid University of Barcelona University of Barcelona Pablo de Olavide University, Sevilla Hospital Doce de Octubre of Madrid University of Castilla La Mancha, Albacete Vall d Hebron University Hospital, Barcelona Autonomous University of Madrid Cajal Institute CSIC; Technical University, Madrid Institute of Health Carlos III, Madrid Catalonian Institute de Bioengineering, Barcelona University of Valencia University of Seville Complutense University of Madrid Bellvitge Institute of Biomedical Research, Barcelona University of Extremadura, Caceres Cavanilles Institute, University of Valencia Autonomous University of Madrid University of Malaga Complutense University of Madrid Institute of Biomedical Research CSIC-UAM, Madrid Marqués de Valdecilla Research Institute, Santander Santa Creu i Sant Pau Hospital, Barcelona University of Santiago de Compostela Center of Applied Medical Research, Univ. Navarra, Pamplona Santa Creu i Sant Pau Hospital, Barcelona Virgen del Rocio University Hospital, University of Sevilla Cajal Institute CSIC, Madrid Biodonostia Research Institute, San Sebastian Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid University of the Basque Country, Bilbao Institute of Biomedical Research IDIBAPS-CSIC, Barcelona Cajal Institute CSIC, Madrid 2016 CIBERNED Annual Report / 11

12 Group Principal investigator Institution Muñoz Cánoves, Pura Naranjo Orovio, José Ramón Navarro Acebes, Xavier Obeso Inchausti, José Ángel Pastor Muñoz, María Asunción Pérez Castillo, Ana María Pérez Tur, Jordi Rodríguez Álvarez, José Rodríguez Díaz, Manuel Sáez Valero, Javier Soriano García, Eduardo Tolosa Sarró, Eduardo Torres Alemán, Ignacio Trullás Oliva, Ramón Vicario Abejón, Carlos Vila Bover, Miquel Vitorica Ferrández, Fco. Javier Wandosell Jurado, Francisco Pompeu Fabra University, ICREA, Barcelona National Center of Biotechnology, CSIC, Madrid Autonomous University of Barcelona Hospitales de Madrid Foundation Center of Applied Medical Research, Univ. Navarra, Pamplona Institute of Biomedical Research CSIC-UAM, Madrid Institute of Biomedicine of Valencia, CSIC Autonomous University of Barcelona University of La Laguna, Tenerife Miguel Hernández University, Elche University of Barcelona Hospital Clinic of Barcelona Cajal Institute CSIC, Madrid Institute of Biomedical Research IDIBAPS-CSIC, Barcelona Cajal Institute CSIC, Madrid Vall d Hebron University Hospital, Barcelona University of Seville Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid PROGRAM 1 PROGRAM 2 53 RESEARCH GROUPS 2016 CIBERNED Annual Report / 12

13 Geographic distribution of CIBERNED research groups CIBERNED Annual Report / 13

14 Organizational structure CIBERNED governing bodies are the Governing Council, the Permanent Commission and the Scientific Director. Governing Council: it is the highest body of CIBERNED. Consists of three representatives of the Carlos III Health Institute, 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 Carlos III Health Institute. 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. Standing Committee: it 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. Scientific Director: 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 Avila 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: The Steering Committee 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. Eduardo Tolosa Sarró Dr. Albert Lleó Bisa Dr. José Javier Lucas Lozano Dr. Adolfo López de Munaín Dra. Teresa Iglesias Vacas Dr. Rafael Fernández Chacón 2016 CIBERNED Annual Report / 14

15 Organization chart Institute of Health Carlos III Governing Council Associated Institutions Standing Committe Steering Committe Scientific Director Manager Deputy Scientific Director Research Program Coordinator Research groups 2016 CIBERNED Annual Report / 15

16 External Scientific Advisory Committee Dr. George Perry Dr. Vincenzo Bonifati Dr. Angel Cedazo-Mínguez Dra. Mary Reilly University of Texas, San Antonio, USA (Chairman) Erasmus University, Rotterdam, The Netherlands Karolinska Institute, Stockholm, Sweden Institute of Neurology, London, United Kingdom 2016 CIBERNED Annual Report / 16

17 Consortium members The Central Administration, represented by: The Institute of Health Carlos III. The Spanish National Research Council (CSIC). The following Autonomous Regions: 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, University Hospital- Research Institute Vall d Hebron Foundation, Bellvitge Biomedical Research Institute Foundation (IDIBELL), Catalonian Institute of Bioengineering, University of Barcelona and Autonomous University of Barcelona. Autonomous Region of Valencia, through the 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 (Doce de Octubre University 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 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. HM Hospitales Research Foundation CIBERNED Annual Report / 17

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19 Scientific report

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21 Research programs From an organizational point of view, CIBERNED research groups are organized around two scientific programs, whose goals, composition and structure are maintained as in previous years, and whose budget allocation is based on the results of the internal scientific evaluation. This organizational structure highlights CIBERNED commitment to encourage translational research and the generation of 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 objective of the structure described is to provide a coherent conceptual framework on which the actual cooperative scientific activity carried out by the different research groups materializes. The Programs are described below: PROGRAM 1 Alzheimer s disease and other degenerative dementias 22 groups 296 researchers PROGRAM 2 Parkinson s disease and other neurodegenerative movement disorders 31 groups 444 researchers 2016 CIBERNED Annual Report / 21

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23 Program 1 Alzheimer s disease and other degenerative dementias Ávila de Grado, Jesús...27 Bullido Gómez-Heras, Mª Jesús...30 Camins Espuny, Antonio...32 Cantero Lorente, José Luis...36 Carro Díaz, Eva...39 Comella Carnicé, Joan Xavier...43 de Felipe Oroquieta, Javier...46 Ferrer Abizanda, Isidro...50 Gutiérrez Pérez, Antonia...56 Lleó Bisa, Alberto...59 López de Ceballos Lafarga, María...64 Matute Almau, Carlos...66 Mengod Los Arcos, Guadalupe...70 Pastor Muñoz, María Asunción...72 de Pedro Cuesta, Jesús...75 Rodríguez Álvarez, José...80 Sáez Valero, Javier...83 Soriano García, Eduardo...86 Torres Alemán, Ignacio...90 Trullás Oliva, Ramón...93 Vitorica Ferrández, Francisco Javier...96 Wandosell Jurado, Francisco...99

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25 Program 1: Alzheimer s disease and other degenerative dementias The aging population and longer life expectancy in our society have led in recent decades to a significant increase in cases of people with Alzheimer s disease (AD) disease, the most common cause of dementia in the elderly and against which there is no effective treatment to date. Currently, more than 500,000 people suffer from this disease in Spain. This figure could quadruple in the next 50 years, with devastating consequences not only for affected individuals and their families but also for the very stability of our health system. The hallmarks of AD are the presence in the patient s brain of two aberrant structures, senile plaques and neurofibrillary tangles, as well as synapse loss (it is considered a synaptopathy), mainly between hippocampal and cortical neurons, and significant neurodegeneration. Some aspects of these pathologies can be reproduced in cellular or animal models. Laboratories around the world work with great interest in identifying new causal and risk genes involved in this pathology that could help clarify its pathophysiological basis and lead to the identification of new therapeutic targets. One of the main problems in the EA is that at the time of clinical diagnosis, the brain has already suffered too extensive and irreparable damage. This requires urgent finding of biomarkers as a way to detect the disease much earlier, even asymptomatic, phases when any therapeutic strategy would have a greater chance of success. In this program, 22 groups of clinical and basic researchers are committed both to the diagnosis and care of patients suffering EA. Thus, research in the laboratory will continue over the next year 2016 combining experience and effort to work in a coordinated manner in the search for new genetic factors, disease biomarkers and new therapeutic strategies in 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. Picture: Antonia Gutiérrez Pérez (415), University of Málaga CIBERNED Annual Report / 25

26 Principal investigator Institution Ávila de Grado, Jesús Bullido Gómez-Heras, Mª Jesús Camins Espuny, Antonio Cantero Lorente, José Luis Carro Díaz, Eva Comella Carnice, 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 Center of Molecular Biology Severo Ochoa CSIC-UAM, Madrid Autonomous University of Madrid University de Barcelona Pablo de Olavide University, Sevilla Hospital Universitario Doce de Octubre, Madrid Foundation Vall d Hebron University Hospital, Barcelona Cajal Institute CSIC, Center of Biomedical Technology, Madrid Institute of Health Carlos III, Madrid Bellvitge Biomedical Research Institute, Barcelona University of Málaga Santa Creu i Sant Pau Hospital, Barcelona Cajal Institute CSIC, Madrid University of the Basque Country, Bilbao IDIBAPS-CSIC Biomedical Research institute, Barcelona Center of Applied Medical Research, University of, Pamplona Autonomous University of Barcelona Miguel Hernández University, Elche University of Barcelona Cajal Institute CSIC, Madrid IDIBAPS-CSIC Biomedical Research institute, CSIC, Barcelona University of Seville Center of Molecular Biology Severo Ochoa CSIC-UAM, Madrid Program 1 is coordinated by Drs. Alberto Lleó (Santa Creu i San Pau Hospital) and Jesús Avila de Grado (Center of Molecular Biology Severo Ochoa CSIC-UAM, Madrid) CIBERNED Annual Report / 26

27 GROUP Jesús Ávila de Grado 401 Principal Investigator Jesús Ávila de Grado Research professor Research team Félix Hernández Pérez UAM professor, Co-PI Marta Bolos Jurado Researcher María Llorens-Martín Researcher Vega García-Escudero Researcher Jerónimo Jurado Arjona Researcher Patricia Martín-Maestro Noemí Pallas Bazarra Jesús Merchán Robira Alberto García Rodríguez Juan Ramón Perea Julia Terreros Roncal Raquel Cuadros Catalán Technician Esther García García Technician Nuria de la Torre Alonso Administrative assistant Centro de Biología Molecular Severo Ochoa C/ Nicolás Cabrera, 1 (Laboratorio 208) Campus de Cantoblanco Madrid (Spain) Tel.: Fax: javila@cbm.csic.es 2016 CIBERNED Annual Report / 27

28 Summary During the last year, we have found, among other studies, that in the adult neurogenesis process, taking place at the dentate gyrus; the overexpression of GSK3, in the stem cells (lacking tau protein), resulted in an increase in the proliferation of those stem cells, in that transgenic mice. However, when the overexpression of GSK3 occurs in neuronal cells (containing tau protein), a degeneration of dentate gyrus and memory impairment were found. It may indicate the role of tau presence when GSK3 is overexpressed. Additionally, a novel tau function was described, also related to the neurons of the dentate gyrus using a tau k.o. mouse model. We have found that tau could regulate the effect of external, negative or positive stimulus, in adult neurogenesis. On the other hand, we have described a novel technique to validate somatic mutations that are only present in the brain of Alzheimer disease patients. That technique involves the use of restriction nucleases acting on DNA lacking the somatic mutations but not on DNA having it. In that way, mutated DNA culd be enriched. This work was done in collaboration with the group of Dr. Soriano (CIBERNED). In addition, we have carried out other works with the groups of Dr. Lucas and de Felipe, both of them members of CIBERNED. Keywords Tau, GSK3, Alzheimer disease, Neurogenesis Publications Soler H., Dorca-Arevalo J., Gonzalez M., Rubio S.E., Avila J., Soriano E. et al. The GABAergic septohippocampal connection is impaired in a mouse model of tauopathy. Neurobiology of Aging. 2017;49: Epub Gomez-Ramos A., Picher A.J., Garcia E., Garrido P., Hernandez F., Soriano E. et al. Validation of Suspected Somatic Single Nucleotide Variations in the Brain of Alzheimer s Disease Patients. Journal of Alzheimer s Disease. 2017;56(3): Epub Gamir-Morralla A, López-Menéndez C, Medina M, Iglesias T. A novel neuroprotection target with distinct regulation in stroke and Alzheimer s disease. In: Neuroprotection in Alzheimer s Disease. Gozes I, editors. Elsevier, ISBN: Garcia-Ayllon M.-S., Botella-Lopez A., Cuchillo-Ibanez I., Rabano A., Andreasen N., Blennow K. et al. HNK-1 Carrier Glycoproteins Are Decreased in the Alzheimer s Disease Brain. Molecular Neurobiology. 2016; Fernandez-Nogales M., Santos-Galindo M., Merchan-Rubira J., Hoozemans J.J.M., Rabano A., Ferrer I. et al. Tau-positive nuclear indentations in P301S tauopathy mice. Brain Pathology Avila J., Pallas N., Bolos M., Sayas C.L., Hernandez F. Intracellular and extracellular microtubule associated protein tau as a therapeutic target in Alzheimer disease and other tauopathies. Expert Opinion on Therapeutic Targets. 2016; 1-9. Torres-Cruz F.M., Rodriguez-Cruz F., Escobar-Herrera J., Barragan-Andrade N., Basurto-Islas G., Ripova D. et al. Expression of Tau Produces Aberrant Plasma Membrane Blebbing in Glial Cells Through RhoA-ROCK-Dependent F-Actin Remodeling. Journal of Alzheimer s Disease. 2016;52(2): Pallas-Bazarra N., Jurado-Arjona J., Navarrete M., Esteban J.A., Hernandez F., Avila J. et al. Novel function of Tau in regulating the effects of external stimuli on adult hippocampal neurogenesis. EMBO Journal Sanchez-Mut J.V., Heyn H., Vidal E., Moran S., Sayols S., Delgado-Morales R. et al. Human DNA methylomes of neurodegenerative diseases show common epigenomic patterns. Translational Psychiatry. 2016;6(1) CIBERNED Annual Report / 28

29 Program 1 Group: Jesús Ávila de Grado Ramirez-Rios S., Denarier E., Prezel E., Vinit A., Stoppin-Mellet V., Devred F. et al. Tau antagonizes end-binding protein tracking at microtubule ends through a phosphorylation-dependent mechanism. Molecular Biology of the Cell. 2016;27(19): Moreno H., Morfini G., Buitrago L., Ujlaki G., Choi S., Yu E. et al. Tau pathology-mediated presynaptic dysfunction. Neuroscience. 2016;325: Llorens-Martin M., Teixeira C.M., Jurado-Arjona J., Rakwal R., Shibato J., Soya H. et al. Retroviral induction of GSK-3β expression blocks the stimulatory action of physical exercise on the maturation of newborn neurons. Cellular and Molecular Life Sciences. 2016; Houck A.L., Hernandez F., Avila J. A simple model to study tau pathology. Journal of Experimental Neuroscience. 2016;10(1): Leon-Espinosa G., Garcia E., Gomez-Pinedo U., Hernandez F., DeFelipe J., Avila J. Decreased adult neurogenesis in hibernating Syrian hamster. Neuroscience. 2016;333: Jurado-Arjona J., Llorens-Martin M., Avila J., Hernandez F. GSK3β overexpression in dentate gyrus neural precursor cells expands the progenitor pool and enhances memory skills. Journal of Biological Chemistry. 2016;291(15): Medina M, Hernández F, Avila J. New Features about Tau Function and Dysfunction. Biomolecules. 2016;6(2). Rabano A., Cuadros R., Merino-Serrais P., Rodal I., Benavides-Piccione R., Gomez E. et al. Protocols for monitoring the development of tau pathology in Alzheimer s disease. Methods in Molecular Biology. 2016;1303: Perez M., Cuadros R., Hernandez F., Avila J. Secretion of full-length tau or tau fragments in a cell culture model. Neuroscience Letters. 2016;634: Fernandez-Blazquez M.A., Avila-Villanueva M., Maestu F., Medina M. Specific Features of Subjective Cognitive Decline Predict Faster Conversion to Mild Cognitive Impairment. Journal of Alzheimer s Disease. 2016;52(1): 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 groups: G111, G401, G104, G307, G409, G412. Other CIBER s collaboration: Yes (CIBERER). Type: CC.AA. Funding agency: Community of Madrid. Funding: Duration: Code I2016/02. Title: Monitoring the onset and evolution of neuronal dysfunctions in propagative neural disorders using microfluidic devices and translational approaches. Principal Investigator: José Antonio del Río. CIBERNED s collaboration: Yes. CIBERNED groups: G504, G401, G403, G111, G307. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 29

30 GROUP María Jesús Bullido Gómez-Heras 510 Principal Investigator María Jesús Bullido Research team Ana Frank García Associate professor Jesús Aldudo Soto Researcher María Recuero Vicente Researcher Ángel Martín Montes Researcher Isabel Sastre Merlín Researcher Henrike Kristen Patricia Llorente Ginés Centro de Biología Molecular Severo Ochoa (CSIC-UAM) Universidad Autónoma de Madrid Edificio CBM Tel.: Fax: CIBERNED Annual Report / 30

31 Summary To identify genes and mechanisms involved in the neurodegeneration process characteristic of Alzheimer s disease (AD), we have developed cellular models simulating different aspects of AD pathogenesis. These models are intended to identify novel genes/ processes involved in AD, that could constitute therapeutic targets, by means of the study of gene expression in the models complemented with genetic association studies in case-control samples. Currently, we work on models of oxidative stress (OS) and of herpes simplex type 1 virus (HSV-1) infection. These models present markers characteristic of AD, like the alterations in amyloid precursor protein (APP) traffic, metabolism and proteolysis and in tau protein phosphorylation. The study of the autophagy lysosome pathway in these models has shown us that infection with HSV-1 and OS profoundly affect the final steps of the route, causing an increased cellular burden of lysosomes accompanied by a significant reduction in the activity of different cathepsins and decreased degradation of lysosomal substrates. Furthermore, we found that another alpha herpesvirus with neurotropic properties, HSV-2, is also able to induce the main neurodegeneration markers previously observed for HSV-1, supporting the potential of diverse infectious agents to participate in the neurodegeneration associated to AD. Together, results of the functional, gene expression and genetic association studies reinforce the hypothesis that a failure of the lysosomal function is a relevant neurodegeneration mechanism, both in the models and in AD patients, so we are currently centered on the analysis of this functional pathway, extending the study to new models that include ipscs derived from patients and controls skin fibroblasts. We also participate in several collaborative projects searching for novel AD genetic risk factors, mainly as part of the Dementia Genetics Spanish Consortium (DEGESCO), as well as with the EADI and IGAP consortia, actively revealing novel factors and functions potentially relevant in the pathogenesis of AD. Keywords Alzheimer, Herpesvirus, oxidative stress, neurodegeneration, lysosomal function, functional genomics, genetic association Publications Itzhaki R.F., Lathe R., Balin B.J., Ball M.J., Bearer E.L., Braak H. et al. Microbes and Alzheimer s disease. Journal of Alzheimer s Disease. 2016;51(4): Sanchez-Benavides G., Pena-Casanova J., Casals-Coll M., Gramunt N., Manero R.M., Puig- Pijoan A. et al. One-Year reference norms of cognitive change in Spanish old adults: Data from the NEURONORMA Sample. Archives of Clinical Neuropsychology. 2016;31(4): Lubrini G, Ríos Lago M, Periañez JA, Tallón Barranco A, De Dios C, Fernández-Fourier M, Diez Tejedor E, Frank García A. The contribution of depressive symptoms to slowness of information processing in relapsing remitting multiple sclerosis. Multiple Sclerosis Oct; 22(12): Research projects Code: SAF R. Title:Identificación de alteraciones funcionales en modelos patogénicos de la enfermedad de Alzheimer: implicacion de la vía lisosomal. Principal Investigator: María J. Bullido Gómez-Heras. CIBERNED groups: G510. Type: National Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 31

32 GROUP Antonio Camins Espuny 402 Principal Investigator Antoni Camins Espuny Full professor Research team Merce Pallás Lliberia Full professor Jordi Vilaplana Associate professor Francesc Sureda Associate professor Carme Pelegrí Associate professor Jaume Folch Associate professor Carme Auladell Associate professor Anna María Canudas Teixidó Lecturer Andrés Jiménez Assistant professor Ester Verdaguer Cardona Lecturer Miren Ettcheto Researcher Sonia Abad Researcher Christian Grañe Farre Assistant professor Itsaso Cabezón Rodríguez Research fellow Victor Rimbau Researcher Oriol Busquets Assistant professor Facultad de Farmacia Departamento de Farmacología y Química Terapéutica Universidad de Barcelona Av. Joan XXIII, s/n Barcelona (Spain) Tel.: camins@ub.edu Elisabet Augé Research fellow 2016 CIBERNED Annual Report / 32

33 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. Our main current lines of research are: The role of obesity and insulin resistance as a target for the prevention of Alzheimer s disease. It is also intended to study the administration of different drugs through nanoparticles to increase its passage through the blood-brain barrier and decrease its toxicity in the treatment of Alzheimer s disease. It is also collaborating in the development of new synthesis molecules for a potential treatment of Alzheimer s disease. Characterization of epigenetic mechanisms involved in Alzheimer s disease. To study the role of MAPK kinases in excitotoxicity processes. To study in cerebral aging in mice the progressive appearance and expansion of degenerative granular structures frequently referred to as PAS granules due to their positive staining with periodic acid-schiff (PAS). PAS granules are present mainly in the hippocampus, although they have also been described in other brain areas such as the piriform and entorhinal cortex, and have been observed in mice, rats and monkeys. PAS granules have been identified as a wide 2016 CIBERNED Annual Report / 33

34 range of brain deposits related to numerous neurodegenerative diseases, such as amyloid deposits, neurofibrillary tangles, Lafora bodies, and polyglucosal bodies. Keywords Alzheimer s, APPSwe / PS1dE9, SAMP8, Resveratrol, aging, diabetes Publications Cosin-Tomas M., Antonell A., Llado A., Alcolea D., Fortea J., Ezquerra M. et al. Plasma mir- 34a-5p and mir-545-3p as Early Biomarkers of Alzheimer s Disease: Potential and Limitations. Molecular Neurobiology. 2016; Corpas R., Revilla S., Ursulet S., Castro-Freire M., Kaliman P., Petegnief V. et al. SIRT1 Overexpression in Mouse Hippocampus Induces Cognitive Enhancement Through Proteostatic and Neurotrophic Mechanisms. Molecular Neurobiology. 2016; Sanchez-Lopez E., Egea M.A., Cano A., Espina M., Calpena A.C., Ettcheto M. et al. PEGylated PLGA nanospheres optimized by design of experiments for ocular administration of dexibuprofenin vitro, ex vivo and in vivo characterization. Colloids and Surfaces B: Biointerfaces. 2016;145: Gonzalez-Castillo C., Ortuno-Sahagun D., Guzman-Brambila C., Marquez-Aguirre A.L., Raisman-Vozari R., Pallas M. et al. The absence of pleiotrophin modulates gene expression in the hippocampus in vivo and in cerebellar granule cells in vitro. Molecular and Cellular Neuroscience. 2016;75: Grizzanti J., Lee H.-G., Camins A., Pallas M., Casadesus G. The therapeutic potential of metabolic hormones in the treatment of age-related cognitive decline and Alzheimer s disease. Nutrition Research. 2016;36(12): Abad S., Ramon C., Pubill D., Camarasa J., Camins A., Escubedo E. Adolescent exposure to MDMA induces dopaminergic toxicity in substantia nigra and potentiates the amyloid plaque deposition in the striatum of APPswe/PS1dE9 mice. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2016;1862(9): Grinan-Ferre C., Palomera-Avalos V., Puigoriol-Illamola D., Camins A., Porquet D., Pla V. et al. Behaviour and cognitive changes correlated with hippocampal neuroinflammaging and neuronal markers in female SAMP8, a model of accelerated senescence. Experimental Gerontology. 2016;80: Folch J., Petrov D., Ettcheto M., Abad S., Sanchez-Lopez E., Garcia M.L. et al. Current Research Therapeutic Strategies for Alzheimer s Disease Treatment. Neural Plasticity. 2016;2016. Grinan-Ferre C., Puigoriol-Illamola D., Palomera-avalos V., Perez-Caceres D., Companys- Alemany J., Camins A. et al. Environmental enrichment modified epigenetic mechanisms in SAMP8 mouse hippocampus by reducing oxidative stress and inflammaging and achieving neuroprotection. Frontiers in Aging Neuroscience. 2016;8(OCT). Griñán-Ferré C, Sarroca S, Ivanova A, Puigoriol-Illamola D, Aguado F, Camins A et al. Epigenetic mechanisms underlying cognitive impairment and Alzheimer disease hallmarks in 5XFAD mice. Aging. 2016;8(4). Ettcheto M., Petrov D., Pedros I., Alva N., Carbonell T., Beas-Zarate C. et al. Evaluation of neuropathological effects of a high-fat diet in a presymptomatic Alzheimer s disease stage in APP/PS1 mice. Journal of Alzheimer s Disease. 2016;54(1): Pedros I, Patraca I, Martinez N, Petrov D, Sureda FX, Auladell C et al. Molecular links between early energy metabolism alterations and Alzheimer s disease.frontiers in bioscience (Landmark edition). 2016; CIBERNED Annual Report / 34

35 Program 1 Group: Antonio Camins Espuny Manich G., Cabezon I., Auge E., Pelegri C., Vilaplana J. Periodic acid-schiff granules in the brain of aged mice: From amyloid aggregates to degenerative structures containing neoepitopes. Ageing Research Reviews. 2016;27: Research projects Code: PI2016/01. Title: Alteraciones del metabolismo glucolipídico y desarrollo de la demencia de alzhéimer. Principal Investigator: Ignacio Torres Alemán. CIBERNED s collaboration: Yes. CIBERNED groups: G409; G402; G511; G502; G412. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: RTC Title: Evaluación de la eficacia de inhibidores epigenéticos en modelos experimentales de patologías humanas. Principal Investigator: Merce Pallas (G402)/ Esther Pérez (G301). CIBERNED s collaboration: Yes. CIBERNED groups: G402; G301. Type: National. Funding agency: Ministerio de Economía, Insdustria y Competitividad. Funding: ,25. Duration: 2016 Code: BFU P. Title: Caracterización de un neo-epitopo cerebral relacionado con el envejecimiento y estudio de la presencia de anticuerpos naturales anti-neoepitopo. Principal Investigator: Carme Pelegri Gavalda. CIBERNED groups: Other CIBER s collaborarion: No. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: SAF C2-1-R. Title: Estudio de la Epoxido hidrolasa soluble como una nueva diana farmacológica para la enfermedad de Alzheimer: modulación del estrés oxidativo y la función mitocondrial. Principal Investigator: Merce Pallas. CIBERNED groups: G402. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: BFU P. Title: Estudio de la presencia de neoepitopos en estructuras degenerativas cerebrales y de la existencia en el plasma de anticuerpos naturales dirigidos contra dichos neo-epitopos. Principal Investigator: Carme Pelegri Gabalda. CIBERNED groups: G402. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: PhD Dissertations Author: Sònia Abad Florensa. Títle: Estudi dels efectes neuroplàstics i neurotòxics de l MDMA en models experimentals d alteracions del Sistema Nerviós Central. Date: 7 July Supervisor: Antonio Camins Espuny. Author: : Ignacio Pedros. Title: Alteraciones metabólicas en el proceso de amiloidogenesis en el hipocampo de ratones APP/PS1. Date: 26 February Supervisor: Jaume Folch. Author: Christian Griñan Ferre. Title: Participación de la modulación epigenética en modelos murinos de envejecimiento y enfermedad de Alzheimer. Date: 12 Dicembre Supervisor: Mercedes Pallas Lliberia CIBERNED Annual Report / 35

36 GROUP José Luis Cantero Lorente 511 Principal Investigator José Luis Cantero Lorente Research team Mercedes Atienza Ruiz Researcher Mayely Sánchez Espinosa Ana María Escorza Aguilar Engineer Sofía Rodríguez Peñuela Technician Juan Jesús Toro Murillo Technician Maité Crespo García Collaborator David García Solís Collaborator Eulogio Gil Néciga Collaborator Laboratorio de Neurociencia Funcional Servicios Centralizados de Investigación (Edificio 21) Universidad Pablo de Olavide Ctra. de Utrera, Km Sevilla (Spain) Tel.: jlcanlor@upo.es 2016 CIBERNED Annual Report / 36

37 Summary In 2016, we have performed experiments aimed at describing failures of structural/functional cortical connectivity patterns underlying prodromal stages of Alzheimer s disease (AD). We found that levels of high-density lipoprotein (HDL) cholesterol were positively correlated with the strength of cortical synchronization, and increased triglycerides accompanied bilateral atrophy of the precuneus in amci patients (González-Escamilla et al., 2016). By using analysis techniques based on the graph theory, we have described how different levels of the cortical hierarchy are affected in the progression to AD. We showed that the transition from normal aging to AD is associated with less efficient cortical networks, characterized by a regional isolation at global, hemispheric and lobular level as well as by a reorganization of cortical hubs. This organization denotes segregation and impaired integration of cortical systems (Romero-García et al., 2016). We further shown that encoding-induced abnormal alpha desynchronized activity over the posterior cingulate cortex together with the magnitude of amygdala atrophy at baseline showed the best accuracy in predicting AD progression, supporting the relevance of combining EEG oscillations and MRI markers for improving early AD detection (Prieto del Val et al., 2016). Finally, we have investigated if subjective memory complaints (SMC) are associated with structural changes in different hippocampal subregions and/or variations in peripheral levels of amyloid-beta (Abeta 1-42). Results showed that individuals with SMC exhibited lower volumes of CA1, CA4, dentate gyrus, and molecular layer together with higher Abeta 1-42 levels compared with those without SMC. Further analyses showed significant associations between lower volume of the dentate gyrus and both poorer memory performance and higher plasma Abeta 1-42 in SMC subjects. These results suggested that SMC might be a condition associated with cerebral vulnerability in nondemented elderly subjects (Cantero et al., 2016). Keywords Cerebral aging, Biomarkers of preclinical and prodromal stages of Alzheimer s disease, Anatomical and functional connectivity of human neocortex Publications Cantero JL, Zaborszky L, Atienza M. Volume Loss of the Nucleus Basalis of Meynert is Associated with Atrophy of Innervated Regions in Mild Cognitive Impairment.Cerebral cortex (New York, N.Y. : 1991) Crespo-Garcia M., Zeiller M., Leupold C., Kreiselmeyer G., Rampp S., Hamer H.M. et al. Slowtheta power decreases during item-place encoding predict spatial accuracy of subsequent context recall. NeuroImage. 2016;142: Gonzalez-Escamilla G., Atienza M., Garcia-Solis D., Cantero J.L. Cerebral and blood correlates of reduced functional connectivity in mild cognitive impairment. Brain Structure and Function. 2016;221(1): Schott J.M., Crutch S.J., Carrasquillo M.M., Uphill J., Shakespeare T.J., Ryan N.S. et al. Genetic risk factors for the posterior cortical atrophy variant of Alzheimer s disease. Alzheimer s and Dementia. 2016;12(8): Cantero J.L., Iglesias J.E., Van Leemput K., Atienza M. Regional hippocampal atrophy and higher levels of plasma amyloid-beta are associated with subjective memory complaints in nondemented elderly subjects. Journals of Gerontology - Series A Biological Sciences and Medical Sciences. 2016;71(9): Prieto Del Val L., Cantero J.L., Atienza M. Atrophy of amygdala and abnormal memory-related alpha oscillations over posterior cingulate predict conversion to Alzheimer s disease. Scientific Reports. 2016; CIBERNED Annual Report / 37

38 Suarez-Gonzalez A., Lehmann M., Shakespeare T.J., Yong K.X.X., Paterson R.W., Slattery C.F. et al. Effect of age at onset on cortical thickness and cognition in posterior cortical atrophy. Neurobiology of Aging. 2016;44: Suárez-González A, Crutch SJ, Roldán Lora F, Franco-Macías E, Gil-Néciga E. Can patients without early, prominent visual deficits still be diagnosed of posterior cortical atrophy?. Journal of the neurological sciences. 2016;367. Suárez-González A, Crutch SJ, Franco-Macías E, Gil-Néciga E. Neuropsychiatric Symptoms in Posterior Cortical Atrophy and Alzheimer Disease.Journal of geriatric psychiatry and neurology. 2016;29(2). Research projects Code: PI2016/01. Title: Alteraciones del metabolismo glucolipídico y desarrollo de la demencia de alzhéimer. Principal Investigator: Ignacio Torres Alemán. CIBERNED s collaboration: Yes. CIBERNED groups: G409; G402; G511; G502; G412. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: P12-CTS Title: Caracterización de las fases presintomáticas y preclínicas de la enfermedad de Alzheimer mediante marcadores biológicos y de neuroimagen. Principal Investigator: José Luis Cantero Lorente. CIBERNED groups: G511. Type: CC.AA. Funding agency : Junta de Andalucía. Funding: Duration: Code: PSI R. Title: Estudio de las oscilaciones cerebrales EEG relacionadas con la memoria como potencial biomarcador de la enfermedad de Alzheimer. Principal Investigator: Mercedes Atienza Ruiz. CIBERNED groups: G511. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 38

39 GROUP Eva María Carro Díaz 502 Principal Investigator Eva María Carro Díaz Research team Desiree Antequera Tienda Technician Fernando Bartolomé Robledo Research fellow Marta González Sánchez José Antonio Molina Arjona Research staff Julián Benito León Research staff Jesús Hernández Gallego Research staff Rocío Trincado Soriano Statistic technician Alberto Villarejo Galende Research staff David Andrés Pérez Martínez Research staff Alejandro Herrero San Martín Research staff Sara Llamas Velasco Research staff Instituto de Investigación Hospital 12 de Octubre Avenida de Córdoba s/n Madrid Tel.: Fax: CIBERNED Annual Report / 39

40 Summary The scientific activity has focused on the study of the physiopathological mechanisms of neurodegenerative diseases, mainly Alzheimer s disease and associated dementias, and to the identification of new therapeutic targets. Specific activities are: Studies on new risk factors, biomarkers of disease and new therapeutic strategies in Alzheimer s disease and other degenerative dementias. a) Research on biomarkers in biological samples non-invasive. b) Cellular and animals models of Alzheimer s disease and others degenerative c) dementias. Molecular pathology in Alzheimer s disease. d) Mechanisms of neurodegeneration, neuroprotection, designing new therapies and drug testing in experimental models. Epidemiological research in the field of neurodegenerative diseases, with special attention to risk factors. a) Publication of results of cohort NEDICES-I. b) Implementation of the cohort NEDICES-II. Clinical research in dementia. a) b) Determination of the clinical utility of biomarkers of recent introduction. Development of new molecular biomarkers and imaging. Keywords Alzheimer disease, dementia, biomarkers, transgenic mice, choroid plexus, therapeutic drugs, amyloid, mild cognitive impairment Publications Haubenberger D., Abbruzzese G., Bain P.G., Bajaj N., Benito-Leon J., Bhatia K.P. et al. Transducerbased evaluation of tremor. Movement Disorders Llamas-Velasco S., Sierra-Hidalgo F., Llorente-Ayuso L., Herrero-San Martin A., Villarejo Galende A., Bermejo Pareja F. Inter-Rater Agreement in the Clinical Diagnosis of Cognitive Status: Data from the Neurological Disorders in Central Spain 2 Pilot Study. Neuroepidemiology. 2016; Benito-Leon J. The Future of Statins: Essential Tremor. Neuroepidemiology. 2016; Agúndez JA, García-Martín E, Martínez C, Benito-León J, Millán-Pascual J, Díaz-Sánchez M et al. Heme Oxygenase-1 and 2 Common Genetic Variants and Risk for Multiple Sclerosis.Scientific reports. 2016;6. Tavares E., Antequera D., Lopez-Gonzalez I., Ferrer I., Minano F.J., Carro E. Potential Role of Aminoprocalcitonin in the Pathogenesis of Alzheimer Disease. American Journal of Pathology. 2016;186(10): Labiano-Fontcuberta A., Benito-Leon J. Radiologically isolated syndrome: An update on a rare entity. Multiple Sclerosis. 2016;22(12): Ferrer I., Garcia-Esparcia P., Carmona M., Carro E., Aronica E., Kovacs G.G. et al. Olfactory receptors in non-chemosensory organs: The nervous system in health and disease. Frontiers in Aging Neuroscience. 2016;8. Louis E.D., Benito-Leon J., Faust P.L. Essential tremor seems to be a risk factor for Parkinson s disease. Parkinsonism and Related Disorders Contador I, Stern Y, Bermejo-Pareja F, Sánchez-Ferro Á, Benito-León J. Is educational attainment associated with increased risk of mortality in people with dementia?. A population-based study. Current Alzheimer research Contador I., del Ser T., Llamas S., Villarejo A., Benito-Leon J., Bermejo-Pareja F. Impact of literacy and years of education on the diagnosis of dementia: A population-based study. Journal of Clinical and Experimental Neuropsychology. 2016; CIBERNED Annual Report / 40

41 Program 1 Group: Eva María Carro Díaz Benito-Leon J., Contador I., Mitchell A.J., Domingo-Santos A., Bermejo-Pareja F. Performance on specific cognitive domains and cause of death: A prospective population-based study in Non-Demented Older Adults (NEDICES). Journal of Alzheimer s Disease. 2016;51(2): Contador I, Almondes K, Fernández-Calvo B, Boycheva E, Puertas-Martín V, Benito-León J et al. Semantic Verbal Fluency: Normative Data in Older Spanish Adults From NEDICES Population- Based Cohort.Archives of clinical neuropsychology : the official journal of the National Academy of Neuropsychologists Bermejo-Pareja F, Llamas-Velasco S, Villarejo-Galende A. Alzheimer s disease prevention: A way forward.revista clinica espanola Benito-Leon J., Domingo-Santos A. Comments on Role of intestinal microbiota in the development of multiple sclerosis. Neurologia Contador I., Bermejo-Pareja F., Fernandez-Calvo B., Boycheva E., Tapias E., Llamas S. et al. The 37 item Version of the Mini-Mental State Examination: Normative Data in a Population-Based Cohort of Older Spanish Adults (NEDICES). Archives of Clinical Neuropsychology. 2016;31(3): Labiano-Fontcuberta A., Mato-Abad V., Alvarez-Linera J., Hernandez-Tamames J.A., Martinez- Gines M.L., Aladro Y. et al. Gray matter involvement in radiologically isolated syndrome. Medicine (United States). 2016;95(13):e Labiano-Fontcuberta A., Mato-Abad V., Alvarez-Linera J., Hernandez-Tamames J.A., Martinez- Gines M.L., Aladro Y. et al. Normal-appearing brain tissue analysis in radiologically isolated syndrome using 3 T MRI. Medicine (United States). 2016;95(27). Sardinero-Garcia C., Santiago-Saez A., Bravo M.D.C., Perea-Perez B., Albarran-Juan M.E., Labajo-Gonzalez E. et al. Responsibility for the loss of opportunity in malignant cancer care in the Spanish public healthcare system. Gaceta Sanitaria. 2016;30(6): Benito-Leon J., Louis E.D., Mato-Abad V., Dydak U., Alvarez-Linera J., Hernandez-Tamames J.A. et al. In vivo neurometabolic profiling in orthostatic tremor. Medicine (United States). 2016;95(37). Benito-Leon J., Louis E.D., Manzanedo E., Hernandez-Tamames J.A., Alvarez-Linera J., Molina- Arjona J.A. et al. Resting state functional MRI reveals abnormal network connectivity in orthostatic tremor. Medicine (United States). 2016;95(29). Benito-Leon J., Contador I., Louis E.D., Cosentino S., Bermejo-Pareja F. Education and risk of incident dementia during the premotor and motor phases of essential tremor (NEDICES). Medicine (United States). 2016;95(33). Benito-Leon J., Domingo-Santos A. Reply to «Orthostatic tremor secondary to recreational use of solvents». Neurologia Matarazzo M., Galan Sanchez-Seco V., Mendez-Guerrero A.J., Gata-Maya D., Domingo- Santos A., Ruiz-Morales J. et al. Drug-Related Eyelid Nystagmus: Two Cases of a Rare Clinical Phenomenon Related to Carbamazepine and Derivatives. Clinical Neuropharmacology Benito-Leon J., Domingo-Santos A. Comment on the case report entitled Secondary orthostatic tremor in the setting of cerebellar degeneration. Journal of Clinical Neuroscience León Ruiz M, García Soldevilla MA, Vidal Díaz MB, Izquierdo Esteban L, Benito-León J, García- Albea Ristol E. Acute myocardial infarction associated with bilateral subacute cardioembolic stroke in the anterior cerebral artery territory: The hidden face of a new-onset atrial fibrillation. Neurologia (Barcelona, Spain) Domingo-Santos A., Sepulveda M., Matarazzo M., Calleja-Castano P., Ramos-Gonzalez A., Saiz A. et al. Intravenous immunoglobulin therapy in a patient with anti-myelin oligodendrocyte glycoprotein-seropositive neuromyelitis optica. Clinical Neuropharmacology. 2016;39(6): Benito-León J, Domingo-Santos Á. Orthostatic Tremor: An Update on a Rare Entity.Tremor and other hyperkinetic movements (New York, N.Y.). 2016; CIBERNED Annual Report / 41

42 Benito-Leon J., Domingo-Santos A. Gut microflora and multiple sclerosis. Journal of the Neurological Sciences. 2016;368:254-. Morrison P.J., Benito-Leon J. Neurologic features in intermediate allele carriers of Huntington disease. Neurology. 2016;87(6): Research projects Code: PI2016/01. Title: Alteraciones del metabolismo glucolipídico y desarrollo de la demencia de alzhéimer. Principal Investigator: Ignacio Torres Alemán. CIBERNED s collaboration: Yes. CIBERNED groups: G409; G402; G511; G502; G412. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: DTS15/ Title: Evaluación del impacto de la imagen PET de amiloide en el diagnóstico de los pacientes con deterioro cognitivo evaluados por sospecha de alzhéimer. Principal Investigator: Dr. Javier Arbizu. CIBER s collaboration: Yes. CIBERNED groups: G504; G502; G609. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: 2016 Code: RTC Title: Plataforma para el seguimiento de trastornos del movimiento (NETMD). Principal Investigator: Julián Benito León. CIBERNED groups: G502. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: N.D. Duration: Code: PI Title: Inflamacion periferica como factor de riesgo en la patologia de la enfermedad de Alzheimer. Principal Investigator: Eva Carro. CIBERNED groups: G502. Type: National. Funding agency: Instituto de Salud Carlos III. Duration: 2016 PhD dissertations Author: Antonio Martínez Salio. Title: Estudio de la incidencia y mortalidad de la enfermedad cerebrovascular en el anciano. Estudio epidemiológico poblacional en la cohorte Nedice. Date: 28 January Supervisor: Félix Bermejo Pareja CIBERNED Annual Report / 42

43 GROUP Joan Xavier Comella Carnicé 413 Principal Investigator Joan Xavier Comella Carnicé Research team Víctor Yuste Mateos PhD fellow Joaquín López Soriano PhD fellow María José Pérez Soriano PhD fellow Bruna Barneda Zahonero PhD fellow Koen Galenkamp PhD fellow Isabel Calleja Yagüe Elena Coccia Julia Soler Flores Master student VHIR Vall Hebron Institut de Recerca Passeig Vall d Hebron Barcelona (Spain) Tel.: Web: joan.comella@vhir.org 2016 CIBERNED Annual Report / 43

44 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 main objective is to characterize these proteins at the molecular level, their involvement in neuronal differentiation and physiology, and their possible role in different pathologies, mainly neurodegenerative diseases. We are also characterizing molecular partners of these molecules that main explain their mechanisms of action. The knowledge of the molecular basis of these receptor antagonists and the activation of survival signaling pathways could be of high relevance to understand the etiology or to open new therapeutic strategies for the treatment of neurodegenerative diseases. We have recently characterized, in collaboration with different CIBERNED groups, the role of FAIM-L in the development of Alzheimer s disease, both in animal models (APP/PS1) and patients (Carriba et al., 2015). At present we are further characterizing this role of FAIM-L, and with this aim we have started the characterization of the Faim knockout model, through immunohistochemistry, electrophysiological and behavior studies. We also have developed adenoassociated viral vectors, which will allow us to overexpress or downregulate FAIM-L levels in brain, to be used in the murine models of Alzheimer s disease, in order to study the role of FAIM-L in the development or progression of the disease, or its potential protector effect in these models. We have recently characterize new unexpected roles for FAIM-L, since we have observed that it is implicated in the control of some non-apoptotic effects of caspases, such as axonal degeneration and long term depression (LTD), through modulation of XIAP levels (Martínez-Mármol et al., 2016). These events of synaptic plasticity are also modulated in neurodegenerative diseases, thus positioning FAIM-L as a good candidate for the treatment of the disease. We are also characterizing two new isoforms of FAIM, one of them specific from neurons. Our main research lines at present are: 1) 2) 3) To study FAIM-L function in 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, particularly NFkB. To characterize FAIM-L functional partners, particularly Siva-1 and XIAP, 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, including the characterization of two new isoforms, one of them neuron-specific. Keywords Alzheimer, neuroinflammation, death receptors, FAIM, TNF, Fas Publications Planells-Ferrer L., Urresti J., Coccia E., Galenkamp K.M.O., Calleja-Yague I., Lopez-Soriano J. et al. Fas apoptosis inhibitory molecules: More than death-receptor antagonists in the nervous system. Journal of Neurochemistry Bosch C, Masachs N, Exposito-Alonso D, Martínez A, Teixeira CM, Fernaud I et al. Reelin Regulates the Maturation of Dendritic Spines, Synaptogenesis and Glial Ensheathment of Newborn Granule Cells.Cerebral cortex (New York, N.Y. : 1991) Sanchez-Osuna M., Martinez-Escardo L., Granados-Colomina C., Martinez-Soler F., Pascual- Guiral S., Iglesias-Guimarais V. et al. An intrinsic DFF40/CAD endonuclease deficiency impairs oligonucleosomal DNA hydrolysis during caspase-dependent cell death: A common trait in human glioblastoma cells. Neuro-Oncology. 2016;18(7): CIBERNED Annual Report / 44

45 Program 1 Group: Joan Xavier Comella Carnicé Martinez-Marmol R., Barneda-Zahonero B., Soto D., Andres R.M., Coccia E., Gasull X. et al. FAIM-L regulation of XIAP degradation modulates Synaptic Long-Term Depression and Axon Degeneration. Scientific Reports. 2016;6. Bosch C, Martínez A, Masachs N, Teixeira CM, Fernaud I, Ulloa F et al. Corrigendum: FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFPlabeled adult-generated neurons.frontiers in neuroanatomy. 2016;10. Research projects Code: PI2015-2/02. Title: Potencial patológico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal Investigator: Joan Xavier Comella Carnicé. CIBERNED s collaboration: Yes. CIBERNED groups: G413; G415; G204; G108; G411. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: Title: Deciphering the link between astrocyte reactivity and neuronal damage in Alzheimer s disease. Principal Investigator: Elena Galea. CIBERNED s collaboration: Yes. CIBERNED groups: G411, G413, G415. Type: Private. Funding agency: Fundaciò La Marato de TV3. Funding: Duration: Code: 2014 SGR Title: Apoptosi i Neurodegeneracio (GRC). Principal Investigator: Joan Xavier Comella Carnicé. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G413. Type: CC.AA. Funding agency: AGAUR-Generalitat de Catalunya. Funding: Duration: 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 groups: G413 Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: SAF R. Title: Relevancia del antagonista de receptores de muerte, FAIM-L, en la enfermedad de Alzheimer. Principal Investigator: Joan Xavier Comella Carnicé. CIBERNED groups: G413. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 45

46 GROUP Javier de Felipe Oroquieta 403 Principal Investigator Javier de Felipe Oroquieta Research team Lidia Alonso-Nanclares Postdoctoral fellow Ruth Benavides-Piccione Postdoctoral fellow Rodrigo Rodríguez Sánchez CSIC researcher Isabel Fernaud Espinosa Postdoctoral fellow Asta Kastanauskaite Postdoctoral fellow Ángel Merchán-Pérez Postdoctoral fellow Alberto Muñoz Postdoctoral fellow Lorena Valdés Lora Technician Ana Isabel García Ramírez Technician Laboratorio Cajal de Circuitos Corticales, CTB, UPM. Campus de Montegancedo Pozuelo de Alcorcón Madrid (Spain) Tel.: Débora Cano Laiseca Technician Mari Carmen Álvarez Pérez Technician Silvia Tapia González Postdoctoral fellow Gonzalo León Espinosa Postdoctoral fellow Mirian Marín Horcajada Lab technician Alejandro Antón Fernández Liulia Diana Furcila Andrea Santuy Muñoz Guillermo Aparicio Torres Marta Turégano Marta Domínguez Sandra Ostos Fernando López María del Carmen Regalado Marta Montero Concepción Rojo Visiting researcher Miguel Miguens Vázquez Visiting researcher Juncal González Visiting researcher Pilar Flores Romero Research and project manager Montserrat Fernández Bouzo Research and project manager Yago Rodríguez Cela Office manager 2016 CIBERNED Annual Report / 46

47 Summary Microorganization of the normal cerebral cortex and alterations of cortical circuits in brain pathologies. The CCCL principally focuses on the microorganization of the normal cerebral cortex (including hippocampus) in various species (particularly humans) and on the alterations of cortical circuits in epilepsy and Alzheimer disease. These studies are performed through the use of anatomical tracers, high resolution immunocytochemistry and 3D light and electron microscopy. Another major aim is to develop informatics technologies to examine the brain. In particular, the research lines carried out at the CCCL are as follows: Study of the components of the column: Development and validation of software tools for 3D segmentation of cells of the complete cortical column in stacks obtained by confocal microscopy in samples from the somatosensory neocortex. Spatial analysis distribution of the segmented cell to determine the distribution patterns of different cell types. Analysis of the spatial distribution of synapses in the six cortical layers by means of spatial statistical tools. Calculation of the volume fraction occupied by mitochondria in the six cortical layers. Pyramidal cells: Intracellular injections and 3D-reconstruction of pyramidal cells and to analyse the microanatomy of these cells in different cortical areas, layers and species. Generation of high resolution confocal microscopy stacks of images of pyramidal cells to analyze the distribution and morphology of dendritic spines in different cortical areas, layers and species. Intrinsic and extrinsic connectivity of the cortical column: Target identification of cortical synapses (FIB/SEM) to determine the proportion of synapses on dendritic spines and dendritic shafts. Analysis of synaptic sizes (FIB/SEM) and development of computer models to explore the possible relationship between synaptic morphology and physiology. Characterization of the afferent and efferent connections of the hindlimb representation area of the primary somatosensory cortex in P14 rats in tract-tracing experiments. Quantification of cellular and subcellular alterations in Alzheimer s Disease (AD) and the possible influence of these alterations on cognition: Quantification of specific synaptic alterations in the neuropil and in identified 3D reconstructed cortical neurons from areas showing early histopathological changes in AD. Application of systematic methods to human tissue from AD patients. Correlation of quantitative results with cognitive impairment. Keywords Microanatomy, cortical circuits, cerebral cortex, mouse and human brain Publications Bosch C, Masachs N, Exposito-Alonso D, Martínez A, Teixeira CM, Fernaud I et al. Reelin Regulates the Maturation of Dendritic Spines, Synaptogenesis and Glial Ensheathment of Newborn Granule Cells.Cerebral cortex (New York, N.Y. : 1991) Munoz M.-S.-O.-S.-C.D., Antolin-Vallespin M., Tapia-Gonzalez S., Sanchez-Capelo A. Smad3 deficiency inhibits dentate gyrus LTP by enhancing GABAA neurotransmission. Journal of Neurochemistry CIBERNED Annual Report / 47

48 Fernandez-Gonzalez P., Benavides-Piccione R., Leguey I., Bielza C., Larranaga P., DeFelipe J. Dendritic-branching angles of pyramidal neurons of the human cerebral cortex. Brain Structure and Function. 2016; Barth A., Burkhalter A., Callaway E.M., Connors B.W., Cauli B., DeFelipe J. et al. Comment on principles of connectivity among morphologically defined cell types in adult neocortex. Science. 2016;353(6304). Bosch C, Martínez A, Masachs N, Teixeira CM, Fernaud I, Ulloa F et al. Corrigendum: FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFPlabeled adult-generated neurons.frontiers in neuroanatomy. 2016;10. Anton-Sanchez L., Bielza C., Benavides-Piccione R., DeFelipe J., Larranaga P. Dendritic and Axonal Wiring Optimization of Cortical GABAergic Interneurons. Neuroinformatics. 2016;14(4): Rabano A., Cuadros R., Merino-Serrais P., Rodal I., Benavides-Piccione R., Gomez E. et al. Protocols for monitoring the development of tau pathology in Alzheimer s disease. Methods in Molecular Biology. 2016;1303: Broadhead M.J., Horrocks M.H., Zhu F., Muresan L., Benavides-Piccione R., DeFelipe J. et al. PSD95 nanoclusters are postsynaptic building blocks in hippocampus circuits. Scientific Reports. 2016;6. Eyal G., Verhoog M.B., Testa-Silva G., Deitcher Y., Lodder J.C., Benavides-Piccione R. et al. Unique membrane properties and enhanced signal processing in human neocortical neurons. elife. 2016;5(OCTOBER2016). Marquez Neila P., Baumela L., Gonzalez-Soriano J., Rodriguez J.-R., DeFelipe J., Merchan-Perez A. A Fast Method for the Segmentation of Synaptic Junctions and Mitochondria in Serial Electron Microscopic Images of the Brain. Neuroinformatics. 2016;14(2): DeFelipe J., Douglas R.J., Hill S.L., Lein E.S., Martin K.A.C., Rockland K.S. et al. Comments and general discussion on The anatomical problem posed by brain complexity and size: A potential solution. Frontiers in Neuroanatomy. 2016;10. Leon-Espinosa G., Garcia E., Gomez-Pinedo U., Hernandez F., DeFelipe J., Avila J. Decreased adult neurogenesis in hibernating Syrian hamster. Neuroscience. 2016;333: Leguey I, Bielza C, Larrañaga P, Kastanauskaite A, Rojo C, Benavides-Piccione R et al. Dendritic branching angles of pyramidal cells across layers of the juvenile rat somatosensory cortex.the Journal of comparative neurology. 2016;524(13). Rojo C., Leguey I., Kastanauskaite A., Bielza C., Larranaga P., Defelipe J. et al. Laminar Differences in Dendritic Structure of Pyramidal Neurons in the Juvenile Rat Somatosensory Cortex. Cerebral Cortex. 2016;26(6): DeFelipe J. Phospho-Tau and cognitive decline in Alzheimer s disease. Commentary: Tau in physiology and pathology. Frontiers in Neuroanatomy. 2016;10. Toharia P., Robles O.D., Fernaud-Espinosa I., Makarova J., Galindo S.E., Rodriguez A. et al. PyramidalExplorer: A new interactive tool to explore morpho-functional relations of human pyramidal neurons. Frontiers in Neuroanatomy. 2016;9. Mellstrom B., Kastanauskaite A., Knafo S., Gonzalez P., Dopazo X.M., Ruiz-Nuno A. et al. Specific cytoarchitectureal changes in hippocampal subareas in dadream mice. Molecular Brain. 2016;9(1) CIBERNED Annual Report / 48

49 Program 1 Group: Javier de Felipe Oroquieta Research projects Code: PI2016/05. Title: Dream inhibitors and Alzheimer s Disease. Principal Investigator: José Ramón Naranjo Orovio. CIBERNED s collaboration: Yes. CIBERNED groups: G307; G106; G403. Other CIBER a collaboration: No. Type: Intramurales. 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. CIBERNED groups: G204, G403. Type: International. Funding agency: Ecole Polytechnique Federale de Lausanne (Suiza), IBM, Universidad Politecnica de Madrid, Consejo Superior de Investigaciones Cientificas, Espana. Funding: ND. Duration: Code: ZEN Title: The Pyramidal Neuron in Cognition and Alzheimer's Disease. Principal Investigator: Javier de Felipe. CIBERNED groups: G403. Type: International. Funding agency: Alzheimer's Association. Funding: ND. Duration: CIBERNED Annual Report / 49

50 GROUP Isidro Ferrer Abizanda 503 Principal Investigator Isidro Ferrer Abizanda Research team Ester Asó Research staff Marta Barrachina Castilla Research staff Montserrat Olivé Plana Research staff Karina Hernández-Ortega Postdoctoral fellow Franc Llorens Postdoctoral fellow Noemi Vidal Sarró Research staff Pol Andrés Benito Belén Ansoleaga Ávila Mayelín Domínguez González Paula García-Esparcia Anusha Konetti Irene López-González Raissa Camila Alvear Contreras Student Margarida Frau Méndez Student Neuropatología, Servicio Anatomía Patológica y Departamento de Patología y Terapeútica Experimental Universidad de Barcelona, campus Bellvitge C/ Feixa Llarga sn, Hospitalet de Llobregat Barcelona Tel.: Móvil: ifa@gmail.com María Francisca García- Garrido Student Meritxell Puig Pinos Student Luis Alberto Escobar Visiting researcher Alejandra Martínez Visiting researcher Andre Palmeira Visiting researcher Katrin Thüne Visiting researcher Margarita Carmona Murillo Techinician Jesús Moreno Lab technician Ester Bergés Pujol Administrative 2016 CIBERNED Annual Report / 50

51 Summary The main aspects of research are centered on the use of combined omics applied to the identification of altered molecular pathways, clusters, and hubs in the cerebral cortex in aging and neurodegenerative diseases with abnormal protein aggregates. In this line, comparative aspects of inflammatory responses have shown marked disease-, region-, and stage-specific changes in the inflammatory response in aging and diseases such as AD, PD, DLB, ALS, and CJD. This aspect has clinical implications, as future therapeutic strategies must contemplate targets differing, depending on the disease and the stage of the disease. Another point has been the identification of altered synthesis machinery from the nucleolus to the ribosome in AD, PD, and CJD, which has implications in the maintenance of cell functions including maintenance of dendrites and synapses; reduced protein synthesis may be causative of neuronal atrophy. A third point has been centered on the identification of altered mitochondria and energy metabolism in AD, PD, and CJD with disease-specific alterations. Finally, oxidative stress markers and metabolomics have been applied to the analysis of regional vulnerability in old age which may contribute to the development of certain prevalent neurodegenerative diseases. Another group of studies was focused on effects of abnormal proteins such as α-synuclein, tau, β-amyloid, and neuroserpin in human cases, and in animal and cellular models. The majority of studies have been aimed at identifying the molecular neuropathology of neurodegenerative diseases in old age as a continuum of classical neuropathological studies based on morphological methods. Several studies are centered on prion diseases covering signaling mechanisms, and analysis of the effects of different strains and biomarkers. In the same line, we have participated in the organization of the Fifth Iberian Congress of Prions sponsored in part by CIBERNED. We invited and edited the production of a minisymposium on Huntington s disease appearing in Brain Pathology that covers hot aspects of the disease including clinical symptoms and underlying mechanisms of cognitive impairment, neuropathology, altered splicing of key cytoskeletal proteins, and noxious effects of aberrant httg mrna. We have also produced an up-dated review about olfactory and taste receptors in the brain and their alterations in neurodegenerative diseases. The function of these central receptors is still not known. Two papers describe new neurodegenerative entities; the first one is a rare familial tauopathy with complex genomic background. The second refers to ARTAG (Aging-related tau astrogliopathy) which is the result of an international agreement to define an agingrelated tau astrogliopathy whose main elements, thorn-like astrocytes, were the subject of seminal studies in our laboratory a few years ago. Finally, several studies have been carried out in collaboration with other groups of CIBERNED and other countries embracing various conditions such as ALS, schizophrenia, HD, and leukodystrophies. We have continued study of the beneficial effects of cannabinoids in mouse models and have started a financed trial of Sativex in patients with mild cognitive impairment, sponsored by the Alzheimer s Association. We have also participated in a clinical trial with albumina replacement in patients with ALS. Regarding innovation, one member of the group has initiated a project sponsored by Caixa Impuls aimed at the identification of novel biomarkers in AD. Keywords Neuropathology, epigenetic, transcriptomic, proteomic, metabolomic, lipidomic, Alzheimer s, Parkinson s, Creutzfeldt-Jakob disease, Frontotemporal lobe degeneration, Amyotrophic lateral sclerosis, X chromosome-linked adrenoleukodystrophy, muscular diseases Publications Gamir-Morralla A, Belbin O, Fortea J, Alcolea D, Ferrer I, Lleó A et al. Kidins220 Correlates with Tau in Alzheimer s Disease Brain and Cerebrospinal Fluid.Journal of Alzheimer s disease : JAD. 2017;55(4). Epub CIBERNED Annual Report / 51

52 Llorens F., Schmitz M., Ferrer I., Zerr I. CSF biomarkers in neurodegenerative and vascular dementias. Progress in Neurobiology Lopez-Gonzalez I., Viana R., Sanz P., Ferrer I. Inflammation in Lafora Disease: Evolution with Disease Progression in Laforin and Malin Knock-out Mouse Models. Molecular Neurobiology. 2016; Zafar S., Younas N., Correia S., Shafiq M., Tahir W., Schmitz M. et al. Strain-Specific Altered Regulatory Response of Rab7a and Tau in Creutzfeldt-Jakob Disease and Alzheimer s Disease. Molecular Neurobiology. 2016; Kovacs G.G., Ferrer I., Grinberg L.T., Alafuzoff I., Attems J., Budka H. et al. Aging-related tau astrogliopathy (ARTAG): harmonized evaluation strategy. Acta Neuropathologica. 2016;131(1): Fernandez B., Fdez E., Gomez-Suaga P., Gil F., Molina-Villalba I., Ferrer I. et al. Iron overload causes endolysosomal deficits modulated by NAADP-regulated 2-pore channels and RAB7A. Autophagy. 2016;12(9): Llorens F, Thüne K, Schmitz M, Ansoleaga B, Frau-Méndez M A, Cramm M. et a. Identification of new molecular alterations in fatal familial insomnia. Human Molecular Genetics. 2016; 25(12): Dominguez M., De Oliveira E., Odena M.A., Portero M., Pamplona R., Ferrer I. Redox proteomic profiling of neuroketal-adducted proteins in human brain: Regional vulnerability at middle age increases in the elderly. Free Radical Biology and Medicine. 2016;95:1-15. Sanchez-Mut J.V., Heyn H., Vidal E., Moran S., Sayols S., Delgado-Morales R. et al. Human DNA methylomes of neurodegenerative diseases show common epigenomic patterns. Translational Psychiatry. 2016;6(1). Mata A., Urrea L., Vilches S., Llorens F., Thune K., Espinosa J.-C. et al. Reelin Expression in Creutzfeldt- Jakob Disease and Experimental Models of Transmissible Spongiform Encephalopathies. Molecular Neurobiology. 2016; Schmitz M., Dittmar K., Llorens F., Gelpi E., Ferrer I., Schulz-Schaeffer W.J. et al. Hereditary Human Prion Diseases: an Update. Molecular Neurobiology. 2016; Fernandez-Nogales M., Santos-Galindo M., Merchan-Rubira J., Hoozemans J.J.M., Rabano A., Ferrer I. et al. Tau-positive nuclear indentations in P301S tauopathy mice. Brain Pathology Garcia-Esparcia P, Koneti A, Rodríguez-Oroz MC, Lago B, Del Rio JA, Ferrer I. Mitochondrial activity in the frontal cortex area 8 and angular gyrus in Parkinson s disease and Parkinson s disease with dementia.brain pathology (Zurich, Switzerland) Frau-Mendez M.A., Fernandez-Vega I., Ansoleaga B., Blanco Tech R., Carmona Tech M., Antonio del Rio J. et al. Fatal familial insomnia: Mitochondrial and protein synthesis machinery decline in the mediodorsal thalamus. Brain Pathology Pinacho R., Villalmanzo N., Meana J.J., Ferrer I., Berengueras A., Haro J.M. et al. Altered CSNK1E, FABP4 and NEFH protein levels in the dorsolateral prefrontal cortex in schizophrenia. Schizophrenia Research. 2016;177(1-3): Pinacho R., Vila E., Prades R., Tarrago T., Castro E., Ferrer I. et al. The glial phosphorylase of glycogen isoform is reduced in the dorsolateral prefrontal cortex in chronic schizophrenia. Schizophrenia Research. 2016;177(1-3): Ferrer I. Mini-symposium: Huntington s disease - introduction. Brain Pathology. 2016;26(6): Cabre R., Jove M., Naudi A., Ayala V., Pinol-Ripoll G., Gil-Villar M.P. et al. Specific metabolomics adaptations define a differential regional vulnerability in the adult human cerebral cortex. Frontiers in Molecular Neuroscience. 2016;9. Alexander J., Kalev O., Mehrabian S., Traykov L., Raycheva M., Kanakis D. et al. Familial early-onset dementia with complex neuropathologic phenotype and genomic background. Neurobiology of Aging. 2016;42: CIBERNED Annual Report / 52

53 Program 1 Group: Isidro Ferrer Abizanda Cacabelos D., Ayala V., Granado-Serrano A.B., Jove M., Torres P., Boada J. et al. Interplay between TDP-43 and docosahexaenoic acid-related processes in amyotrophic lateral sclerosis. Neurobiology of Disease. 2016;88: Pinacho R, Vila E, Prades R, Tarragó T, Castro E, Ferrer, I, Ramos B. The glial phosphorylase of glycogen isoform is reduced in the dorsolateral prefrontal cortex in chronic schizophrenia. Schizophrenia research. 2016; 177(1): Pinacho R, Villalmanzo N, Meana JJ, Ferrer I, Berengueras A, Haro JM. et al. Altered CSNK1E, FABP4 and NEFH protein levels in the dorsolateral prefrontal cortex in schizophrenia. Schizophrenia research. 2016; 177(1), Ferrer I., Garcia-Esparcia P., Carmona M., Carro E., Aronica E., Kovacs G.G. et al. Olfactory receptors in non-chemosensory organs: The nervous system in health and disease. Frontiers in Aging Neuroscience. 2016;8. Tavares E., Antequera D., Lopez-Gonzalez I., Ferrer I., Minano F.J., Carro E. Potential Role of Aminoprocalcitonin in the Pathogenesis of Alzheimer Disease. American Journal of Pathology. 2016;186(10): Schmitz M, Llorens F, Pracht A, Thom T, Correia Â, Zafar S et al. Regulation of human cerebrospinal fluid malate dehydrogenase 1 in sporadic Creutzfeldt-Jakob disease patients.aging. 2016;8(11). Aso E., Andres-Benito P., Ferrer I. Delineating the Efficacy of a Cannabis-Based Medicine at Advanced Stages of Dementia in a Murine Model. Journal of Alzheimer s Disease. 2016;54(3): Aso E., Andres-Benito P., Carmona M., Maldonado R., Ferrer I. Cannabinoid receptor 2 participates in amyloid-β processing in a mouse model of Alzheimer s disease but plays a minor role in the therapeutic properties of a cannabis-based medicine. Journal of Alzheimer s Disease. 2016;51(2): Lopez-Gonzalez I., Palmeira A., Aso E., Carmona M., Fernandez L., Ferrer I. FOXP2 Expression in Frontotemporal Lobar Degeneration-Tau. Journal of Alzheimer s Disease. 2016;54(2): Lachen-Montes M., Gonzalez-Morales A., de Morentin X.M., Perez-Valderrama E., Ausin K., Zelaya M.V. et al. An early dysregulation of FAK and MEK/ERK signaling pathways precedes the β-amyloid deposition in the olfactory bulb of APP/PS1 mouse model of Alzheimer s disease. Journal of Proteomics. 2016;148: Barrera-Ocampo A., Arlt S., Matschke J., Hartmann U., Puig B., Ferrer I. et al. Amyloid-β precursor protein modulates the sorting of testican-1 and contributes to its accumulation in brain tissue and cerebrospinal fluid from patients with Alzheimer disease. Journal of Neuropathology and Experimental Neurology. 2016;75(9): Ansoleaga B., Garcia-Esparcia P., Llorens F., Hernandez-Ortega K., Carmona M., Del Rio J.A. et al. Altered mitochondria, protein synthesis machinery, and Purine metabolism are molecular contributors to the pathogenesis of Creutzfeldt-Jakob disease. Journal of Neuropathology and Experimental Neurology. 2016;75(8): Lopez-Gonzalez I., Perez-Mediavilla A., Zamarbide M., Carmona M., Escribano B.T., Glatzel M. et al. Limited unfolded protein response and inflammation in neuroserpinopathy. Journal of Neuropathology and Experimental Neurology. 2016;75(2): Aso E, Ferrer I. CB2 Cannabinoid Receptor As Potential Target against Alzheimer s Disease. Frontiers in neuroscience. 2016;10. Lopez Gonzalez I., Garcia-Esparcia P., Llorens F., Ferrer I. Genetic and transcriptomic profiles of inflammation in neurodegenerative diseases: Alzheimer, Parkinson, Creutzfeldt-Jakob and Tauopathies. International Journal of Molecular Sciences. 2016;17(2). Sorolla MA, Rodríguez-Colman MJ, Vall-Llaura N, Vived C, Fernández-Nogales M, Lucas JJ et al. Impaired PLP-dependent metabolism in brain samples from Huntington disease patients and transgenic R6/1 mice.metabolic brain disease. 2016;31(3). Cacabelos D, Ramírez-Núñez O, Granado-Serrano AB, Torres P, Ayala V, Moiseeva V et al. Early and gender-specific differences in spinal cord mitochondrial function and oxidative stress markers in a mouse model of ALS.Acta neuropathologica communications. 2016; CIBERNED Annual Report / 53

54 Ferrer I., Climent F., Vidal N., Escobar L.A., Sanchez J.J., Fernandez-Coello A. 74 Year-Old Woman with Progressive Weakness. Brain Pathology. 2016;26(3): Martinez-Maldonado A., Luna-Munoz J., Ferrer I. Incidental corticobasal degeneration. Neuropathology and Applied Neurobiology. 2016;42(7): Research projects Code: AGL REDT. Title: Red nacional de priones. Principal Investigator: José Antonio del Río. CIBERNED s collaboration: Yes. CIBERNED groups: G114; G509; G503; G609; G207. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: CIBERNED 2014/02. Title: Mecanismos epigenéticos implicados en la etiología y progresión de las demencias neurodegenerativas rápidamente progresivas. Principal Investigator: Miguel Calero. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G601. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: PI2016/04. Title: The ALS CIBERNED Challenge: Accelerating New Drug Discovery. Principal Investigator: Adolfo López De Munain Arregui. CIBERNED s collaboration: Yes. CIBERNED groups: G609, G303, G503, G408. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: FI12/ Title: Ayudas Predoctorales de formación en investigación (PFIS). Principal Investigator: Isidro Ferrer. CIBERNED groups: G503. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: FIS PIE/ Title: Epigenetic and environmental factors bracing cognitive impairment and melancholic depression in elderly and neurodegeneration. Principal Investigator: Isidro Ferrer Abizanda. CIBERNED groups: G503. Type: National. Funding agency: Insituto de Salud Carlos III. Funding: Duration: Code: PI14/ Title: Establecimiento de un algoritmo mitocondrial para predecir el riesgo de progresión a la enfermedad de Alzheimer en pacientes con deterioro cognitivo leve. Principal Investigator: Marta Barrachina. CIBERNED groups: G503. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: ND. Duration: Code: FIS PI/ Title: Interactomas y marcadores para discriminar factores de vulnerabilidad neuronal regional en la neurodegeneración asociada al envejecimiento normal y patológico en el SNC. Principal Investigator: Isidro Ferrer Abizanda. CIBERNED groups: G503. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: CIBERNED Annual Report / 54

55 Program 1 Group: Isidro Ferrer Abizanda Code: N.D. Title: Development of a kit for an early Alzheimer s disease detection. Principal Investigator: Marta Barrachina. CIBERNED groups: G503. Type: Private. Funding agency: Obra social La Caixa. Funding: Duration: 2016 Code: Sat CIEN-02. Title: The part of the cloud challenge on neuroinflammation. SATIVEX. Alzheimer s Association. Principal Investigator: Isidro Ferrer Abizanda. CIBERNED groups: G503. Type: International. Funding agency: Alzheimer s Association. Funding: Duration: PhD dissertations Author: Belén Ansoleaga Ávila. Title: Mitochondria and purine metabolism in Alzheimer s disease and Creutzfeldt-Jakob s disease with a note on biomarkers. Date: 4 November de Supervisor: Isidro Ferrer Abizanda. Author: Paula García Esparza. Title: Identification of a risk transcriptome and proteome in Parkinson s disease, Dementia with Lewy bodies and rapidly progressive Dementia with Lewy bodies. Date: 2 de Dicember de Supervisor: Isidro Ferrer Abizanda CIBERNED Annual Report / 55

56 GROUP Antonia Gutiérrez Pérez 415 Principal Investigator Antonia Gutiérrez Pérez Full professor Research team José Carlos Dávila Cansino Full professor Zafaruddin Khan Associate professor David Baglietto Vargas Assistant professor Raquel Sánchez Varo Postdoctoral researcher Elisabeth Sánchez Mejías CIBERNED postdoctoral researcher Laura Trujillo Estrada Postdoctoral fellow Mercedes Aneiros Ferrer CIBERNED technician Ángela Gómez Arboledas Cristina Núñez Díaz Juan José Fernández Valenzuela Dpto. Biología Celular, Genética y Fisiología (Área de Biología Celular) Facultad de Ciencias, Universidad de Málaga Campus de Teatinos 29071, Málaga (Spain) Tel.: agutierrez@uma.es 2016 CIBERNED Annual Report / 56

57 Summary During 2016, we have continued our research on glial pathology in Alzheimer s disease in collaboration with Dr. Javier Vitorica of the University of Seville. In particular, we have analyzed the microglial response that occurs in transgenic models with Abeta pathology (APP/PS1) or with tau pathology (Thy-tau22), as well as in post-mortem samples of AD patients (hippocampus/parahippocampal gyrus). While in the hippocampus of the Abeta models there is a strong microglial activation associated with the development of the amyloid pathology, in the Tau model the microglial cells do not show activation but instead begin to present a degenerative phenotype at advanced ages. One of the most relevant finding has been the identification in the hippocampus of patients with dementia (Braak V-VI) of a very limited activation of the microglia, along with a marked degenerative process of these cells following a regional gradient (dentate gyrus>ca3>ca1). We have determined that the soluble phospho-tau is the causative agent of this microglial toxicity, which explains that the Abeta models (APP or APP/PS1) do not display this degenerative microglial phenotype. In order to deepen this objective and develop animal models that better recapitulate human pathology, we have pharmacologically (immunosuppression) or genetically (IL4KO, Gal3KO and CSFR1KO) manipulated the microglial population in different transgenic models. Microglial dysfunction results in an aggravation of pathology including an increase in Abeta levels, along with synaptic and neuronal loss. In addition, we have extended our study on microglia to other cortical regions of interest. Another of our objectives during this year, part of a CIBERNED cooperative project coordinated by Dr. Joan Comella, has been to study the pathological potential of astrocytes in transgenic models and patient samples. In connection with this line, we have worked on a project funded by La Marató TV3 to decipher the role of the reactive astrocytes and to identify different functional populations. Finally, we have collaborated with various national and international research groups for the development of other objectives that are also included in our research line on Alzheimer s disease. Keywords Alzheimer, neuroinflammation, neuropathology, microglia, astroglia, hippocampus, transgenic models, patients Publications Sanchez-Mejias E., Navarro V., Jimenez S., Sanchez-Mico M., Sanchez-Varo R., Nunez-Diaz C. et al. Soluble phospho-tau from Alzheimer s disease hippocampus drives microglial degeneration. Acta Neuropathologica. 2016; Hernández-Rodríguez M, Correa-Basurto J, Gutiérrez A, Vitorica J, Rosales-Hernández MC. Asp32 and Asp228 determine the selective inhibition of BACE1 as shown by docking Reserach projects Code: PI2015-2/02. Title: Potencial patológico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal Investigator: Joan X. Comella Carnice. CIBERNED s collaboration: Yes. CIBERNED groups: G413, G415, G204, G108, G411. Type: Intramurales. Funding agency: CIBERNED. Funding: Duratión: CIBERNED Annual Report / 57

58 Code: Title: Deciphering the link between astrocyte reactivity and neuronal damage in Alzheimer s disease. Principal Investigator: Elena Galea. CIBERNED s collaboration: Yes. CIBERNED groups: G411, G413, G415. Type: Private. Funding agency: La Maratò de TV3. Funding: Duration: Code: CTS Title: Oligomerización y toxicidad de los peptidos de Abeta: búsqueda de nuevas dianas de interés terapéutico en la enfermedad de Alzheimer. Principal Investigator: Javier Vitorica. CIBERNED s collaboration: Yes. CIBERNED groups: G411, G415. Type: CC.AA. Funding agency: Junta de Andalucía. Funding: ,5. Duration: Code: FIS PI15/ Title: Evaluando la disfunción microglial y astroglial como base del proceso neurodegenerativo y la demencia en la enfermedad de Alzheimer: nuevas aproximaciones terapéuticas. Principal Investigator: Antonia Gutiérrez Pérez. CIBERNED s collaboration: Yes. CIBERNED groups: G415, G411. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: BFU R. Title: Una estrategia para recuperar y prevenir la pérdida de la memoria Principal Investigator: Zafaruddin Khan. CIBERNED groups: Type: National. Funding agency: Ministerio de Economía y Competitividad. Funding: Duration: CIBERNED Annual Report / 58

59 GROUP Alberto Lleó Bisa 504 Principal Investigator Alberto Lleó Bisa Neurologist, Principal Investigator Research team Rafael Blesa González Neurologist, head of the Neurologist Service Juan Fortea Neurologist Daniel Acolea Rodríguez Neurologist María Carmona Neurologist Jordi Clarimón Echavarría Researcher Olivia Belbin Researcher Isabel Sala Neuropsychologist M. Belén Sánchez-Saudinós Neuropsychologist Andrea Subirana Neuropsychologist Lidia García Nurse Laura Videla Neuropsychologist Roser Ribosa Neurologist Ignacio Illán-Gala Neurologist Estrella Morenas Researcher Martí Colom Oriol Dols Marta Querol Laura Cervera Eduard Vilaplana Jordi Pegueroles Victor Montal Laia Muñoz Lab technician Raúl Núñez Lab technician Servicio de Neurología Hospital Santa Cruz y San Pablo Sant Antoni María Claret Barcelona (España) Tel.: Fax: alleo@santpau.es 2016 CIBERNED Annual Report / 59

60 Summary Our group is composed of a multidisciplinary team of neurologists, neuropsychologists, biologists, engineers and lab technicians. The activities of the group are focused on translational and clinical aspects of neurodegenerative dementias. In 2016 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 continnum, 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. We have implemented novel microscopy techniques (Arrat Tomography combined with STED) that allow the analyses of human brain with high spatial resolution. The group offers a unique environment for clinicians and investigators to tackle basic and translational aspects of neurodegeneration. Keywords Alzheimer, dementia, cerebrospinal fluid, amyloid, imaging, biomarkers Publications Gamir-Morralla A, Belbin O, Fortea J, Alcolea D, Ferrer I, Lleó A et al. Kidins220 Correlates with Tau in Alzheimer s Disease Brain and Cerebrospinal Fluid.Journal of Alzheimer s disease : JAD. 2017;55(4). Epub van Waalwijk van Doorn LJ, Gispert JD, Kuiperij HB, Claassen JA, Arighi A, Baldeiras I et al. Improved Cerebrospinal Fluid-Based Discrimination between Alzheimer s Disease Patients and Controls after Correction for Ventricular Volumes.Journal of Alzheimer s disease : JAD. 2017;56(2). Epub Carmona-Iragui M., Santos T., Videla S., Fernandez S., Benejam B., Videla L. et al. Feasibility of Lumbar Puncture in the Study of Cerebrospinal Fluid Biomarkers for Alzheimer s Disease in Subjects with Down Syndrome. Journal of Alzheimer s Disease. 2017;55(4): Epub Tell-Marti G, Puig-Butille JA, Potrony M, Plana E, Badenas C, Antonell A et al. A Common Variant in the MC1R Gene (p.v92m) is associated with Alzheimer s Disease Risk.Journal of Alzheimer s disease : JAD. 2017;. Epub Pegueroles J, Vilaplana E, Montal V, Sampedro F, Alcolea D, Carmona-Iragui M et al. Longitudinal brain structural changes in preclinical Alzheimer s disease.alzheimer s & dementia : the journal of the Alzheimer s Association Verheijen J., van Den Bossche T., van der Zee J., Engelborghs S., Sanchez-Valle R., Llado A. et al. A comprehensive study of the genetic impact of rare variants in SORL1 in European earlyonset Alzheimer s disease. Acta Neuropathologica. 2016; Suarez-Calvet M., Kleinberger G., Araque Caballero M.A., Brendel M., Rominger A., Alcolea D. et al. strem2 cerebrospinal fluid levels are a potential biomarker for microglia activity in early-stage Alzheimer s disease and associate with neuronal injury markers. EMBO Molecular Medicine Leuzy A, Chiotis K, Hasselbalch SG, Rinne JO, de Mendonça A, Otto M et al. Pittsburgh compound B imaging and cerebrospinal fluid amyloid-β in a multicentre European memory clinic study.brain : a journal of neurology. 2016;139(Pt 9). Mandelli M.L., Vilaplana E., Brown J.A., Hubbard H.I., Binney R.J., Attygalle S. et al. Healthy brain connectivity predicts atrophy progression in non-fluent variant of primary progressive aphasia. Brain. 2016;139(10): CIBERNED Annual Report / 60

61 Program 1 Group: Alberto Lleó Bisa Ferrari R., Wang Y., Vandrovcova J., Guelfi S., Witeolar A., Karch C.M. et al. Genetic architecture of sporadic frontotemporal dementia and overlap with Alzheimer s and Parkinson s diseases. Journal of Neurology, Neurosurgery and Psychiatry Servian-Morilla E., Takeuchi H., Lee T.V., Clarimon J., Mavillard F., Area-Gomez E. et al. A POGLUT1 mutation causes a muscular dystrophy with reduced Notch signaling and satellite cell loss. EMBO Molecular Medicine. 2016;8(11): Sogorb-Esteve A., Garcia-Ayllon M.-S., Fortea J., Sanchez-Valle R., Lleo A., Molinuevo J.-L. et al. Cerebrospinal fluid Presenilin-1 increases at asymptomatic stage in genetically determined Alzheimer s disease. Molecular Neurodegeneration. 2016;11(1):1-11. Spataro N., Roca-Umbert A., Cervera-Carles L., Valles M., Anglada R., Pagonabarraga J. et al. Detection of genomic rearrangements from targeted resequencing data in Parkinson s disease patients. Movement Disorders Cosin-Tomas M., Antonell A., Llado A., Alcolea D., Fortea J., Ezquerra M. et al. Plasma mir- 34a-5p and mir-545-3p as Early Biomarkers of Alzheimer s Disease: Potential and Limitations. Molecular Neurobiology. 2016; Dangla-Valls A., Molinuevo J.L., Altirriba J., Sanchez-Valle R., Alcolea D., Fortea J. et al. CSF microrna Profiling in Alzheimer s Disease: a Screening and Validation Study. Molecular Neurobiology. 2016; 1-8. Cervera-Carles L., Alcolea D., Estanga A., Ecay-Torres M., Izagirre A., Clerigue M. et al. Cerebrospinal fluid mitochondrial DNA in the Alzheimer s disease continuum. Neurobiology of Aging Yanez M.J., Belbin O., Estrada L.D., Leal N., Contreras P.S., Lleo A. et al. c-abl links APP-BACE1 interaction promoting APP amyloidogenic processing in Niemann-Pick type C disease. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2016;1862(11): Kun-Rodrigues C, Ross OA, Orme T, Shepherd C, Parkkinen L, Darwent L et al. Analysis of C9orf72 repeat expansions in a large international cohort of dementia with Lewy bodies.neurobiology of aging van der Zee J, Gijselinck I, Van Mossevelde S, Perrone F, Dillen L, Heeman B et al. TBK1 Mutation Spectrum in an Extended European Patient Cohort with Frontotemporal Dementia and Amyotrophic Lateral Sclerosis.Human mutation Bras J., Djaldetti R., Alves A.M., Mead S., Darwent L., Lleo A. et al. Exome sequencing in a consanguineous family clinically diagnosed with early-onset Alzheimer s disease identifies a homozygous CTSF mutation. Neurobiology of Aging. 2016;46:236.e1-236.e6. Compta Y, Ramos-Campoy O, Grau-Rivera O, Colom-Cadena M, Clarimón J, Martí MJ et al. Conjoint FTLD-FUS of the neuronal intermediate filament inclusion disease type, progressive supranuclear palsy and alzheimer s pathology presenting as parkinsonism with early falls and late hallucinations, psychosis and dementia.neuropathology and applied neurobiology Varga A.W., Wohlleber M.E., Gimenez S., Romero S., Alonso J.F., Ducca E.L. et al. Reduced slow-wave sleep is associated with high cerebrospinal fluid aβ42 levels in cognitively normal elderly. Sleep. 2016;39(11): Pickett E.K., Koffie R.M., Wegmann S., Henstridge C.M., Herrmann A.G., Colom-Cadena M. et al. Non-Fibrillar Oligomeric Amyloid-β within Synapses. Journal of Alzheimer s Disease. 2016;53(3): Sanchez-Benavides G., Pena-Casanova J., Casals-Coll M., Gramunt N., Manero R.M., Puig- Pijoan A. et al. One-Year reference norms of cognitive change in Spanish old adults: Data from the NEURONORMA Sample. Archives of Clinical Neuropsychology. 2016;31(4): Youssif C., Flix B., Belbin O., Comalada M. Measuring NLR oligomerization IV: Using Förster resonance energy transfer (FRET)-fluorescence lifetime imaging microscopy (FLIM) to determine the close proximity of inflammasome components. Methods in Molecular Biology. 2016;1417: Zwan M.D., A. Lleó, J. Fortea, R. Blesa. Erratum: Use of amyloid-pet to determine cutpoints for CSF markers: A multicenter study (Neurology (2016) 86 (50-58)). Neurology. 2016;86(22): CIBERNED Annual Report / 61

62 Parnetti L., Eusebi P., Lleo A. Cerebrospinal Fluid Biomarkers for Target Engagement and Efficacy in Clinical Trials for Alzheimer s and Parkinson s Diseases. Frontiers of Neurology and Neuroscience. 2016;39: Research projects Code: DTS15/ Title: Evaluación del impacto de la imagen PET de amiloide en el diagnóstico de los pacientes con deterioro cognitivo evaluados por sospecha de alzhéimer. Principal Investigator: Dr. Javier Arbizu. CIBERNED s collaboration: Yes. CIBERNED groups: G504, G502, G609. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: 2016 Code: MRTVE/P29. Title: A comprehensive genomic analysis of patients with motor neuron disease and frontotemporal dementia to disentangle the missing genetic architecture of amyotrophic lateral sclerosis. Principal Investigator: Jordi Clarimón. CIBERNED groups: G504. Type: National. Funding agency: Maratón Radio Televisión Española. Funding: Duration: Code: PI14/ Title: Marcadores sinápticos en la enfermedad de Alzheimer. Principal Investigator: Alberto Lleó. CIBERNED groups: G504. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: PI12/ 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 Clarimón. CIBERNED groups: G504. Type: National. Funding agency: Instuto de Salud Carlos III. Funding: Duration: Code: PI15/ Title: Estudio del perfil de expresión de microrna exosomal en biofluidos para la identificación de biomarcadores de uso diagnóstico en la demencia frontotemporal. Principal Investigator: Jordi Clarimón. CIBERNED s groups: G504. Type: National. Funding agency: Instuto de Salud Carlos III. Funding: Duration: 2016 Code: 2014 SGR 235. Title: Grup de Recerca en Demècies: Sant Pau. Principal Investigator: Jordi Clarimón. CIBERNED groups: G504. Type: CC.AA. Funding agency: AGAUR-Generalitat Catalunya. Funding: Duration: 2016 Code: PI15/ Title: Valoración y caracterización de biomarcadores en la fase preclínica de la enfermedad de Alzheimer. Principal Investigator: Olivia Belbín. CIBERNED groups: G504. Type: National. Funding agency: Instuto de Salud Carlos III. Funding: Duration: CIBERNED Annual Report / 62

63 Program 1 Group: Alberto Lleó Bisa PhD dissertations Author: Oriol Dols Icardo. Title: Assessing the genetic overlap in amyotrophic lateral sclerosis and frontotemporal dementia. Date: 15 Dicember de Supervisor: Jordi Clarimón Echavarría. Author: Marta Marquié Sayagués. Title: Validación en tejido cerebral humano de [F-18]-AV-1451 (T807), un nuevo marcador de la proteína tau-phf para tomografía por emisión de positrones. Date: 22 de November de Supervisor: Alberto Lleó Bisa. Author: Marc Suárez Calvet. Title: Degeneración lobular frontotemporal: estudio clínico, neuropatológico y de biomarcadores. Date: 24 de November de Supervisor: Alberto Lleó Bisa CIBERNED Annual Report / 63

64 GROUP María López de Ceballos Lafarga 506 Prinicipal Investigator María López de Ceballos Lafarga Tenure Scientist, CSIC Research team Clara González Martínez Researcher Alba Garcimartín Álvarez Student Nora Kadiri Moreno Student Sahba Seddhigi Student Instituto Cajal, CSIC Dr. Arce, Madrid (Spain) Tel.: Fax: CIBERNED Annual Report / 64

65 Summary Alzheimer Disease (AD) is characterized by aberrant protein accumulation like β-amyloid and neurofibrillary tangles, by neuronal death and by neuroinflammation. This neuroinflammation is caused by glial cell (astrocytes and microglia) activation that will release inflammatory substances. However, alternative microglial activation may be a possible mechanism to counteract neuroinflammation, by increasing the release of anti-inflammatory substances. On the other hand, it is known that an increase in inflammation caused by recurrent infections enhances the risk of suffering AD, and some systemic infections worsen cognitive impairment of AD patients. Herpes Simplex Virus Type 1 (HSV-1) is one of those infections known to increase the risk for suffering the neurologic condition. An enhancement in neuroinflammation due to infections is a possible mechanism that may explain such increased risk. Therefore, in this project the evolution of levels in glial cell markers, pro- and anti-inflammatory parameters in frontal cortex of control subjects and patients categorized by their Braak stages have been analyzed. In addition, the possible relationship between neuroinflammation in AD and viral infection was investigated. The levels of different proteins were assessed by Western blotting in frontal cortex from control subjects, devoid of pathology, and AD patients, as defined in Braak stages (Neurologic Tissue Brain Bank, Bellevitge Hospital, Barcelona). HSV-1 titre was measured by PCR. Indeed, there was a statistically significant increase in cyclooxygenase-2, the astrocyte marker GFAP, the anti-inflammatory cytokine IL4 and HSV-1 titre. No changes were observed in arginase-1, characteristic of M2 alternative activation, or the microglial cell marker Iba1. A positive and significant correlation was observed between GFAP and IL4 (p=0.0003), that suggests an astrocytic origin rather than microglial. On the other hand, a positive correlation between age and GFAP levels (p=0.036), and between age and HSV-1 (p=0.0041) was found. Our results show a large increase in neuroinflammation in the latest Braak stages which could be related with the HSV-1 infection. Keywords Alzheimer disease, braak stages, infection, HSV-1, Neuroinflammation Publications Kofalvi A., Lemos C., Martin-Moreno A.M., Pinheiro B.S., Garcia-Garcia L., Pozo M.A. et al. Stimulation of brain glucose uptake by cannabinoid CB2 receptors and its therapeutic potential in Alzheimer s disease. Neuropharmacology. 2016; 110: Pajares M., Jimenez-Moreno N., Garcia-Yague A.J., Escoll M., de Ceballos M.L., Van Leuven F. et al. Transcription factor NFE2L2/NRF2 is a regulator of macroautophagy genes. Autophagy. 2016; Navarro-Dorado J., Villalba N., Prieto D., Brera B., Martin-Moreno A.M., Tejerina T. et al. Vascular dysfunction in a transgenic model of Alzheimer s disease: Effects of CB1R and CB2R cannabinoid agonists. Frontiers in Neuroscience. 2016; 10:422. Research projects Code: BFU C2-1-R Title: Alteraciones cerebrales inducidas por la dieta: diferencias sexuales y mecanismos protectores de las hormonas gonadales y la Tibolona. Principal Investigator: Luis Miguel García Segura/María A. Arévalo Arévalo CIBERNED groups: G506. Other CIBER s collaboration: CIBEROBN. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 65

66 GROUP Carlos Matute Almau 404 Principal Investigator Carlos Matute Almau Full professor Research team 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 Researcher associate Asier Ruiz Núñez Postdoctoral fellow Mónica Benito Tania Quintela Andrea Manterola Juaristi Alazne Zabala Olaizola Carolina Ortiz Sanz Jon Gejo Crispín Maripaz Serrano Regal Department of Neurosciences Faculty of Medicine and Dentistry University of País Vasco Leioa (Spain) Tel.: Fax: carlos.matute@ehu.es Ainara Martínez González Technician CIBERNED Saioa Marcos Ezquerro Technician CIBERNED Juan Carlos Chara Technician CIBERNED 2016 CIBERNED Annual Report / 66

67 Summary The main goal of our laboratory is to unveil the pathological mechanisms of glial cells in Alzheimer disease (AD) and to find glial therapeutic targets. In the last year, we have described a 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 turn, we have observed that the expression levels of key intermediaries in that pathway (integrin β1, NOX2, GFAP and others) increase in astrocytes located close and distant to amyloid plaques during AD progression. Electron microscope data in APP/PS1/tau transgenic mice shows that these markers are located in astrocytes which are severely damaged, display perisynaptic changes and hypertrophia. These results suggest that Aβ1-42 may cause primary astrogliosis in AD and alter the tripartite synapse and astrocyte volume. On the other hand, we have observed that Aβ1-42 favors oligodendrocyte maturation and myelin repair after demyelination via activation of Fyn kinase, and that there is substantial hypermyelination in the APP/PS1/tau triple transgenic mice as well as in AD postmortem brain. In addition, the myelin sheath is thicker while compound action potentials have lower amplitude and slower conduction velocity which may underlie cognitive deficits in AD. Lastly, we examined propagation of pathogenic human α-synuclein in Lewy bodies from Parkinson s disease brains using microfluidics. Initial results indicate that this protein is taken up by neurons and astroctyes, and transported intracellularly, being astrocytes resistant to its toxicity. In addition, we have observed neuronal apoptosis folliwing astrocyte exposure to Lewy bodies. These findings point to a relevant role of astrocytes in disease propagation. Therefore, we are generating human astrocytes differentiated form ipscs derived from fibroblasts of patients with sporadic Parkinson disease and with LRRK2 mutations. Keywords β-amyloid toxicity, α-synuclein, astrocytes, oligodendrocytes, myelin, oxidative stress Publications van der Zee J, Gijselinck I, Van Mossevelde S, Perrone F, Dillen L, Heeman B et al. TBK1 Mutation Spectrum in an Extended European Patient Cohort with Frontotemporal Dementia and Amyotrophic Lateral Sclerosis.Human mutation Wyssenbach A., Quintela T., Llavero F., Zugaza J.L., Matute C., Alberdi E. Amyloid β-induced astrogliosis is mediated by β1-integrin via NADPH oxidase 2 in Alzheimer s disease. Aging Cell Verkhratsky A., Matteoli M., Parpura V., Mothet J.-P., Zorec R. Astrocytes as secretory cells of the central nervous system: Idiosyncrasies of vesicular secretion. EMBO Journal Yi C., Mei X., Ezan P., Mato S., Matias I., Giaume C. et al. Astroglial connexin43 contributes to neuronal suffering in a mouse model of Alzheimer s disease. Cell Death and Differentiation. 2016;23(10): Abiega O, Beccari S, Diaz-Aparicio I, Nadjar A, Layé S, Leyrolle Q et al. Neuronal Hyperactivity Disturbs ATP Microgradients, Impairs Microglial Motility, and Reduces Phagocytic Receptor Expression Triggering Apoptosis/Microglial Phagocytosis Uncoupling.PLoS biology. 2016;14(5). Saab A.S., Tzvetavona I.D., Trevisiol A., Baltan S., Dibaj P., Kusch K. et al. Oligodendroglial NMDA Receptors Regulate Glucose Import and Axonal Energy Metabolism. Neuron. 2016;91(1): Forostyak O, Forostyak S, Kortus S, Sykova E, Verkhratsky A, Dayanithi G. Physiology of Ca(2+) signalling in stem cells of different origins and differentiation stages.cell calcium. 2016;59(2-3). Gomis-Gonzalez M., Matute C., Maldonado R., Mato S., Ozaita A. Possible therapeutic doses of cannabinoid type 1 receptor antagonist reverses key alterations in fragile X syndrome mouse model. Genes. 2016;7(9) CIBERNED Annual Report / 67

68 Borralleras C., Mato S., Amedee T., Matute C., Mulle C., Perez-Jurado L.A. et al. Synaptic plasticity and spatial working memory are impaired in the CD mouse model of Williams-Beuren syndrome. Molecular Brain. 2016;9(1). Verkhratsky A, Rodríguez-Arellano JJ, Parpura V, Zorec R. Astroglial calcium signalling in Alzheimer s disease.biochemical and biophysical research communications Sole-Domenech S., Cruz D.L., Capetillo-Zarate E., Maxfield F.R. The endocytic pathway in microglia during health, aging and Alzheimer s disease. Ageing Research Reviews. 2016;32: Benito-Munoz M., Matute C., Cavaliere F. Adenosine A1 receptor inhibits postnatal neurogenesis and sustains astrogliogenesis from the subventricular zone. GLIA. 2016;64(9): Zorec R., Parpura V., Verkhratsky A. Astroglial Vesicular Trafficking in Neurodegenerative Diseases. Neurochemical Research. 2016; Dayanithi G., Verkhratsky A. Calcium signalling in stem cells: Molecular physiology and multiple roles. Cell Calcium Kortus S., Srinivasan C., Forostyak O., Ueta Y., Sykova E., Chvatal A. et al. Physiology of spontaneous [Ca2+]i oscillations in the isolated vasopressin and oxytocin neurones of the rat supraoptic nucleus. Cell Calcium Peng L., Li B., Verkhratsky A. Targeting astrocytes in bipolar disorder. Expert Review of Neurotherapeutics. 2016; 1-9. Arellano RO, Sánchez-Gómez MV, Alberdi E, Canedo-Antelo M, Chara JC, Palomino A et al. Axon-to-Glia Interaction Regulates GABAA Receptor Expression in Oligodendrocytes.Molecular pharmacology. 2016;89(1). Soria F.N., Zabala A., Pampliega O., Palomino A., Miguelez C., Ugedo L. et al. Cystine/glutamate antiporter blockage induces myelin degeneration. GLIA. 2016;64(8): Plattner H., Verkhratsky A. Inseparable tandem: Evolution chooses atp and ca2+ to control life, death and cellular signalling. Philosophical Transactions of the Royal Society B: Biological Sciences. 2016;371(1700). Cavaliere F., Benito-Munoz M., Matute C. Organotypic Cultures as a Model to Study Adult Neurogenesis in CNS Disorders. Stem Cells International. 2016;2016. Verkhratsky A, Zorec R, Rodriguez JJ, Parpura V. Pathobiology of neurodegeneration: The role for astroglia.opera medica et physiologica. 2016;1. Domercq M, Szczupak B, Gejo J, Gómez-Vallejo V, Padro D, Gona KB et al. PET Imaging with [(18)F]FSPG Evidences the Role of System xc(-) on Brain Inflammation Following Cerebral Ischemia in Rats.Theranostics. 2016;6(11). Rose C.R., Verkhratsky A. Principles of sodium homeostasis and sodium signalling in astroglia. GLIA. 2016;64(10): Illes P., Verkhratsky A. Purinergic neurone-glia signalling in cognitive-related pathologies. Neuropharmacology. 2016;104: Kortus S, Srinivasan C, Forostyak O, Zapotocky M, Ueta Y, Sykova E et al. Sodium-calcium exchanger and R-type Ca(2+) channels mediate spontaneous [Ca(2+)]i oscillations in magnocellular neurones of the rat supraoptic nucleus.cell calcium. 2016;59(6). Verkhratsky A., Nedergaard M. The homeostatic astroglia emerges from evolutionary specialization of neural cells. Philosophical Transactions of the Royal Society B: Biological Sciences. 2016;371(1700) CIBERNED Annual Report / 68

69 Program 1 Group: Carlos Matute Almau Research projects Code: IT Title: Bases moleculares y celulares de la neurodegeneración. Principal Investigator: Carlos Matute Almau. CIBERNED groups: G404. Type: CC.AA. Funding agency: Gobierno Vasco. Funding: Duration: Code: Coen Pathfinder Title: Protection of neurons in vitro and in vivo from Synuclein toxicity by molecular tweezers. Principal Investigator: Erwan Bezard. CIBERNED groups: G404. Type: European. Fundig agency: COEN Pathfinder. Funding: Duration: Code: SAF R. Title: Therapeutic potential of neurotransmitter receptors in glial cells. Principal Investigator: Carlos Matute Almau. CIBERNED groups: G404. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: Merck KGaA. Title: Could reeducating microglia by P2X4R manipulation constitute an alternative therapy for MS?. Principal Investigator: María Domercq Garcia. CIBERNED groups: G404. Type: International. Funding agency: Merck KGaA. Funding: Duration: Code: ARSEP2015. Title: Evaluating the potential of targeting 2-AG hydrolytic enzymes MAGL and ABHD6 as novel therapeutic strategy in multiple sclerosis. Principal Investigator: Carlos Matute Almau. CIBERNED groups: G404. Type: International. Funding agency: ARSEP Foundation. Funding: Duration: Code: Title: Neurogenic potential of dimethylfumarate. Principal Investigator: Carlos Matute Almau. CIBERNED groups: G404. Type: International. Funding agency: Biogen Pharma. Funding: Duration: Code: WOK Title: Evaluating the Therapeutic Potential of Endocannabinoid Hydrolysis Inhibitors in Myelin Diseases. Principal Investigator: Susana Mato. CIBERNED groups: G404. Type: Private. Funding agency: WOK Foundation. Funding: Duration: 2016 PhD dissertations Author: Mónica Benito Muñoz. Title: The role of purinergic receptor A1 in neurogenesis modulation from the subventricular zone. Date: 17 June Supervisor: Fabio Cavaliere. Author: Manuel Canedo Antelo. Title: Interacción entre quinasas y moléculas apoptóticas en la muerte oligodendroglial excitotóxica. Date: 19 February Supervisor: María Victoria Sánchez Gómez CIBERNED Annual Report / 69

70 GROUP Guadalupe Mengod Los Arcos 508 Principal Investigator Guadalupe Mengod los Arcos Full professor Research team Roser Cortés Research staff Mª Teresa Vilaró Research staff Rocío Martín-Álvarez Lab technician Silvia Serrano Researcher Marina Di Bari Postdoctoral fellow Giovanni Di Pinto Master student Meritxell Pintó Juncà Master student Travis M. Stewart Master student Anna Martínez Sirés Visiting student Theodora Zymatoura Visiting student Noah Valverde Offermann Visiting student Stephen G. Mclnerney Visiting student IIBB-CSIC, IDIBAPS, CIBERNED Tel.: Fax: CIBERNED Annual Report / 70

71 Summary Our group is composed of two sub-lines, which have as main and common theme neurodegeneration. During 2016 we have continued to investigate the relationship between camp and the cellular and molecular aspects of neurodegenerative diseases with inflammatory component. The results we have obtained have shown the possible influence of camp on the evolution of EAE in mice, when chronically treated with PDE7 inhibitors. We have described a novel PDE7 inhibitor that significantly reduces the clinical symptoms of these mice in a similar manner to that produced by treatment with fingolimod, a drug used in multiple sclerosis therapy. We have studied in the spinal cord of these treated mice the mrna expression of PDE4 and PDE7 as well as several cellular markers involved in inflammatory processes. In addition, we have continued to collaborate with the group of Dr. Tata of the Sapienza University of Rome in which we have analyzed the cholinergic system in the spinal cord of EAE mice by in situ hybridization and by histochemical enzymatic reactions. We continue to collaborate with the group of Dr. de Gortari of the group of Molecular Physiology of the INPRFM of Mexico DF. We are analyzing in a triple transgenic mouse model of Alzheimer s disease the effect of the stimulation of these receptors on amyloid Aβ peptide deposition and on other neuropathological and behavioral alterations observed in this animal model. Keywords Neurodegeneration, neuroinflammation, molecular neuropharmacology Publications Martin-Alvarez R., Paul-Fernandez N., Palomo V., Gil C., Martinez A., Mengod G. A preliminary investigation of phoshodiesterase 7 inhibitor VP3.15 as therapeutic agent for the treatment of experimental autoimmune encephalomyelitis mice. Journal of Chemical Neuroanatomy. 2017;80: Epub Valdes-Moreno M.I., Alcantara-Alonso V., Estrada-Camarena E., Mengod G., Amaya M.I., Matamoros-Trejo G. et al. Phosphodiesterase-7 inhibition affects accumbal and hypothalamic thyrotropin-releasing hormone expression, feeding and anxiety behavior of rats. Behavioural Brain Research. 2017;319: Epub Di Bari M, Di Pinto G, Reale M, Mengod G, Tata AM. Cholinergic system and neuroinflammation: Implication in multiple sclerosis.central nervous system agents in medicinal chemistry Cortés R, Vilaró MT, Mengod G. Visualization of 5-HT Receptors Using Radioligand-Binding Autoradiography.Current protocols in pharmacology. 2016; CIBERNED Annual Report / 71

72 GROUP María A. Pastor Muñoz 507 Principal Investigator María Asunción Pastor Muñoz Research team Pau Pastor Muñoz Co-Principal Investigator Miquel Aguilar Barberá MD, PhD Martín Martínez Villar PhD Maite Aznarez Sanado PhD Elkin Luis García PhD Javier Bernacer María PhD Gonzalo Arrondo PhD María Asunción Fernández Seara PhD Elisa Mengual Poza PhD Silvia Romero Contreras Nurse Juan Pablo Tartari MD Luis Eudave Ramos Silvia Zarraluqui López Sara Ortega Cubero Laboratorio de Neuroimagen Center for Applied Medical Research Universidad de Navarra Pamplona, España Tel.: Fax: CIBERNED Annual Report / 72

73 Summary The group of Maria Pastor/Pau Pastor (Neuroimaging and Neurogenetics Laboratories, Center for Applied Medical Research, CIMA, University of Navarra) is working on projects focused to the identification of diagnostic markers that would help to understand and personalize neurodegenerative diseases. The Group is working actively on genetic and neuroimaging markers in dementia and parkinsonism throughout magnetic resonance and next generation sequencing. In 2016 the research of the group has been very relevant. For instance, there are 4 on-going PhD projects from which 15 articles has been published. To highlight the identification of specific MRI patterns linked to specific genetic variants in dementia. Currently, the projects focus on the influence of genetic variants in the evolution of dementias, novel patterns of amyloid- PET and genetic and cerebrospinal fluid markers. In addition, we are working on the identification of novel genetic risk variants for Parkinson disease and Essentials tremor. Dr. Pau Pastor performed a Postdoctoral research in Washington University (St. Louis, MO, USA), under the direction of Dr. Alison Goate. In 2005 he started his own Group that currently accounts for 4 people. He has a large number of international and national collaborations. He is one of the founders of the Spanish genetic consortium for dementia (DEGESCO). Keywords Neuroimaging, functional magnetic resonance imaging, ASL, genetics, SNPs, Gen Publications Giri A., Mok K.Y., Jansen I., Sharma M., Tesson C., Mangone G. et al. Lack of evidence for a role of genetic variation in TMEM230 in the risk for Parkinson s disease in the Caucasian population. Neurobiology of Aging. 2017;50:167.e e13. Epub Jiménez-Jiménez FJ, García-Martín E, Alonso-Navarro H, Martínez C, Zurdo M, Turpín-Fenoll L et al. Thr105Ile (rs ) polymorphism in the histamine-1-methyl-transferase (HNMT) gene and risk for restless legs syndrome.journal of neural transmission (Vienna, Austria : 1996). 2017;124(3). Epub Verheijen J., van Den Bossche T., van der Zee J., Engelborghs S., Sanchez-Valle R., Llado A. et al. A comprehensive study of the genetic impact of rare variants in SORL1 in European earlyonset Alzheimer s disease. Acta Neuropathologica. 2016; Ferrari R., Wang Y., Vandrovcova J., Guelfi S., Witeolar A., Karch C.M. et al. Genetic architecture of sporadic frontotemporal dementia and overlap with Alzheimer s and Parkinson s diseases. Journal of Neurology, Neurosurgery and Psychiatry Müller SH, Girard SL, Hopfner F, Merner ND, Bourassa CV, Lorenz D et al. Genome-wide association study in essential tremor identifies three new loci.brain : a journal of neurology. 2016;139(Pt 12). Benitez B.A., Davis A.A., Jin S.C., Ibanez L., Ortega-Cubero S., Pastor P. et al. Resequencing analysis of five Mendelian genes and the top genes from genome-wide association studies in Parkinson s Disease. Molecular Neurodegeneration. 2016;11(1). Kun-Rodrigues C, Ross OA, Orme T, Shepherd C, Parkkinen L, Darwent L et al. Analysis of C9orf72 repeat expansions in a large international cohort of dementia with Lewy bodies.neurobiology of aging Fernandez-Seara M.A., Rodgers Z.B., Englund E.K., Wehrli F.W. Calibrated bold fmri with an optimized ASL-BOLD dual-acquisition sequence. NeuroImage. 2016;142: Agúndez JA, García-Martín E, Martínez C, Benito-León J, Millán-Pascual J, Díaz-Sánchez M et al. Heme Oxygenase-1 and 2 Common Genetic Variants and Risk for Multiple Sclerosis.Scientific reports. 2016; CIBERNED Annual Report / 73

74 van der Zee J, Gijselinck I, Van Mossevelde S, Perrone F, Dillen L, Heeman B et al. TBK1 Mutation Spectrum in an Extended European Patient Cohort with Frontotemporal Dementia and Amyotrophic Lateral Sclerosis.Human mutation Martinez M., Valencia M., Vidorreta M., Luis E.O., Castellanos G., Villagra F. et al. Trade-off between frequency and precision during stepping movements: Kinematic and BOLD brain activation patterns. Human Brain Mapping Eudave L., Aznarez-Sanado M., Luis E.O., Martinez M., Fernandez-Seara M.A., Pastor M.A. Motor sequence learning in the elderly: differential activity patterns as a function of hand modality. Brain Imaging and Behavior. 2016; Luis E., Ortiz A., Eudave L., Ortega-Cubero S., Borroni B., Van Der Zee J. et al. Neuroimaging Correlates of Frontotemporal Dementia Associated with SQSTM1 Mutations. Journal of Alzheimer s Disease. 2016;53(1): Jimenez-Jimenez F.J., Garcia-Martin E., Alonso-Navarro H., Lorenzo-Betancor O., Ortega- Cubero S., Pastor P. et al. A family study of DRD3 rs6280, SLC1A2 rs and MAPT rs variants in essential tremor. Neurological Research. 2016; 1-8. Pan X., Qian T., Fernandez-Seara M.A., Smith R.X., Li K., Ying K. et al. Quantification of liver perfusion using multidelay pseudocontinuous arterial spin labeling. Journal of Magnetic Resonance Imaging. 2016;43(5): Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Research projects Code: SAF R. Title: Identificación de genes implicados en enfermedad de Parkinson y temblor esencial mediante secuenciacion de nueva generacion y analisis de haplotipos. Principal Investigator: Pau Pastor. CIBERNED groups: G507. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 74

75 GROUP Jesús de Pedro Cuesta 509 Principal Investigator Jesús de Pedro Cuesta Head of unit Research team Enrique Alcalde Cabrero Javier Almazán Isla Technician Fuencisla Avellanal Calzadilla Technician Miguel Calero Lara Researcher lab head Olga Calero Rueda Researcher fellow Javier Damián Moreno Researcher fellow Belén Frades Payo Technician Pablo Martínez Martín Staff researcher María del Carmen Rodríguez Blázquez Researcher María Ruiz Tovar Researcher fellow Juana San Emeterio Mardomingo Lab technician Carmen Matesanz Blanco Lab technician Alexandra Moreno García Researcher Centro Nacional de Epidemiología Instituto de Salud Carlos III Monforte de Lemos Madrid, España Tel.: Fax: jpedro@isciii.es 2016 CIBERNED Annual Report / 75

76 Summary Public Health/aging Subgroup In the field of epidemiology and etiology of neurodegenerative diseases, the most important contribution with the participation of the three subgroups is the publication of the model of drivers. This model (from which an article on incidence was published the previous year) combines molecular elements, clinical and general epidemiology and animal models, providing a transversal view of sporadic neurodegenerative pathologies, suggesting interpretations that encompass other endocrine and vascular conformational pathologies, and the idea of continuity between certain neurodegenerations from clinical onset to advanced age. Regarding aging and disability, several articles deal with the analysis of confinement related to disability and with the loss of key functions, mainly mental and secondly musculoskeletal. Finally, a consensus article among Spanish professionals points out the characteristics of the relationship between mental health and public health, as well as the priorities in prevention and surveillance in this field. According to data from the surveillance of human transmissible spongiform encephalopathies, no new cases of variant Creutzfeldt-Jakob disease (vcjd) have been diagnosed. Commissioned by the Spanish drug agency (AEMPS), a study suggests that Guillain-Barré syndrome does not appear to be related to influenza H1N1 vaccine. A system of pharmacovigilance in Multiple Sclerosis is designed at the request of the AEMPS. Results subgroup During the past decade, since the development and validation of instruments to evaluate nonmotor manifestations of Parkinson s Disease, which have provided information on the importance of these symptoms, it has begun to consider this disease as a complex multisystem disorder. However, a critical review of Non-Motor Symptoms Scale showed shortcomings and implementation problems that have prompted a new version, whose validation study (sponsored by the International Parkinson and Movement Disorder Society (IPMDS) began in 2016). In this year and with the support of the IPMDS, several systematic reviews of scales for assessment of posture, gait and balance, and disability in Parkinson s disease and behavioral disorders in Huntington s disease have been carried out. Several recommended scales were identified for predicted use in disability for Parkinson s disease and behavioral disorders in Huntington, but the development of a specific scale for gait and balance in Parkinson s disease was recommended. We have also carried-out several studies on quality of life in different populations, transcultural adaptation of questionnaires, global assessment scales for Parkinson s disease, analysis of properties and performance of different evaluation scales for neuropsychological disorders, and analysis of the effects of stereotactic surgery on non-motor symptoms in Parkinson s disease. Molecular Biology Subgroup The current objectives of this subgroup are: 1) 2) 3) Molecular diagnosis. Basic research on genetic susceptibility factors in Alzheimer s disease and prion diseases. Molecular basis of conformational diseases, including AD and CJD. During 2016, a close collaboration with different CIBERNED groups has been maintained in three collaborative projects: the DEGESCO consortium, a CIBERNED collaborative project, and the National Prion Network (AGL REDT Network of Excellence). In the context of the DEGESCO consortium, a number of shared or differential genetic risk factors have been studied in different neurodegenerative diseases such as Alzheimer s disease and Creutzfeldt-Jakob disease. In the collaborative project of CIBERNED initiated in 2015 on Epigenetic mechanisms involved in the etiology and progression of rapidly progressive Neurodegenerative Dementias, our group through in silico approaches, route analysis or genetic variations indicative of genetic hotspots, has studied the hypermutability of the CpG dinucleotides as a pathogenic mechanism common to various neurodegenerative diseases, focusing as an object of study in Alzheimer, Parkinson and Creutzfeldt-Jakob. Several polymorphisms (some of them in coding regions) have been identified in different genes associated with methylation and error correction (TDG, PCNA, MBD4, GADD45A and APOBEC1) that appear to be associated with an alteration in the function of the protein they encode, and with an increased risk for neurodegenerative diseases. The group also collaborates with Biocross, SL., in the pursuit of early biomarkers for Alzheimer s disease (metabolomics markers), and with the company AMO Pharma Ltd, in the exploration of treatment response markers (tideglusib) for juvenile-onset myotonic dystrophy. On 2016 CIBERNED Annual Report / 76

77 Program 1 Group: Jesús de Pedro Cuesta the other hand, the group collaborates with other institutions that provide a fundamental support, highlighting the collaboration with the CIEN Foundation for the early diagnosis of Alzheimer s disease (Vallecas project, and the University of the República de Uruguay in the study of peripheral expression of CNS neurodegenerative diseases. Keywords Alzheimer s disease, Parkinson s disease, Creutzfeldt-Jakob disease, Huntington s disease, epidemiology, etiology, neurodegeneration, aging, disability, assessment scales, non-motor symptoms, systematic review, conformational diseases, genetic risk factors, biomarkers, metabolomics Publications Biundo R., Weis L., Fiorenzato E., Gentile G., Giglio M., Schifano R. et al. Erratum to Double-blind Randomized Trial of tdcs Versus Sham in Parkinson Patients With Mild Cognitive Impairment Receiving Cognitive Training Brain Stimulation 8 (2015) Brain Stimulation Minikel E.V., Vallabh S.M., Lek M., Estrada K., Samocha K.E., Sathirapongsasuti J.F. et al. Quantifying prion disease penetrance using large population control cohorts. Science Translational Medicine. 2016;8(322). Van Der Heeden J.F., Marinus J., Martinez-Martin P., Rodriguez-Blazquez C., Geraedts V.J., Van Hilten J.J. Postural instability and gait are associated with severity and prognosis of Parkinson disease. Neurology. 2016;86(24): Martinez-Martin P., Leentjens A.F.G., de Pedro-Cuesta J., Chaudhuri K.R., Schrag A.E., Weintraub D. Accuracy of screening instruments for detection of neuropsychiatric syndromes in Parkinson s disease. Movement Disorders. 2016;31(3): Bloem B.R., Marinus J., Almeida Q., Dibble L., Nieuwboer A., Post B. et al. Measurement instruments to assess posture, gait, and balance in Parkinson s disease: Critique and recommendations. Movement Disorders. 2016;31(9): Shulman L.M., Armstrong M., Ellis T., Gruber-Baldini A., Horak F., Nieuwboer A. et al. Disability Rating Scales in Parkinson s Disease: Critique and Recommendations. Movement Disorders. 2016;31(10): Mestre T.A., van Duijn E., Davis A.M., Bachoud-Levi A.-C., Busse M., Anderson K.E. et al. Rating scales for behavioral symptoms in Huntington s disease: Critique and recommendations. Movement Disorders. 2016;31(10): Linares C., Martinez-Martin P., Rodriguez-Blazquez C., Forjaz M.J., Carmona R., Diaz J. Effect of heat waves on morbidity and mortality due to Parkinson s disease in Madrid: A time-series analysis. Environment International. 2016;89-90:1-6. van der Heeden J.F., Marinus J., Martinez-Martin P., van Hilten J.J. Evaluation of severity of predominantly non-dopaminergic symptoms in Parkinson s disease: The SENS-PD scale. Parkinsonism and Related Disorders. 2016;25:39-44 de Pedro-Cuesta J., Martinez-Martin P., Rabano A., Ruiz-Tovar M., Alcalde-Cabero E., Calero M. Etiologic framework for the study of neurodegenerative disorders as well as vascular and metabolic comorbidities on the grounds of shared epidemiologic and biologic features. Frontiers in Aging Neuroscience. 2016;8. Villar-Vazquez R., Padilla G., Fernandez-Acenero M.J., Suarez A., Fuente E., Pastor C. et al. Development of a novel multiplex beads-based assay for autoantibody detection for colorectal cancer diagnosis. Proteomics. 2016;16(8): Rizos A., Sauerbier A., Antonini A., Weintraub D., Martinez-Martin P., Kessel B. et al. A European multicentre survey of impulse control behaviours in Parkinson s disease patients treated with shortand long-acting dopamine agonists. European Journal of Neurology. 2016;23(8): CIBERNED Annual Report / 77

78 Klingelhoefer L., Rizos A., Sauerbier A., McGregor S., Martinez-Martin P., Reichmann H. et al. Night-time sleep in Parkinson s disease the potential use of Parkinson s KinetiGraph: a prospective comparative study. European Journal of Neurology. 2016;23(8): De Pedro-Cuesta J., Martinez-Martin P., Rabano A., Alcalde-Cabero E., Jose Garcia Lopez F., Almazan-Isla J. et al. Drivers: A biologically contextualized, cross-inferential view of the epidemiology of neurodegenerative disorders. Journal of Alzheimer s Disease. 2016;51(4): Damian J., Pastor-Barriuso R., Valderrama-Gama E., de Pedro-Cuesta J. Association of detected depression and undetected depressive symptoms with long-term mortality in a cohort of institutionalised older people ERRATUM. Epidemiology and Psychiatric Sciences. 2016; 1-. Kohnen R., Martinez-Martin P., Benes H., Trenkwalder C., Hogl B., Dunkl E. et al. Rating of daytime and nighttime symptoms in RLS: Validation of the RLS-6 scale of restless legs syndrome/willis- Ekbom disease. Sleep Medicine. 2016;20: Kohnen R, Martinez-Martin P, Benes H, Trenkwalder C, Högl B, Dunkl E et al. Validation of the Kohnen Restless Legs Syndrome-Quality of Life instrument.sleep medicine. 2016;24. Gunther R., Richter N., Sauerbier A., Chaudhuri K.R., Martinez-Martin P., Storch A. et al. Nonmotor symptoms in patients suffering from motor neuron diseases. Frontiers in Neurology. 2016;7. Gil-Prieto R., Pascual-Garcia R., San-Roman-Montero J., Martinez-Martin P., Castrodeza-Sanz J., Gil-De-Miguel A. Measuring the burden of hospitalization in patients with Parkinson s disease in Spain. PLoS ONE. 2016;11(3). Damian J., Pastor-Barriuso R., Valderrama-Gama E., de Pedro-Cuesta J. Association of detected depression and undetected depressive symptoms with long-term mortality in a cohort of institutionalised older people. Epidemiology and Psychiatric Sciences. 2016; Aguera-Ortiz L., Hernandez-Tamames J.A., Martinez-Martin P., Cruz-Orduna I., Pajares G., Lopez- Alvarez J. et al. Structural correlates of apathy in Alzheimer s disease: A multimodal MRI study. International Journal of Geriatric Psychiatry Wijers IG, Ayala A, Rodriguez-Blazquez C, Rodriguez-Laso A, Rodriguez-Rodriguez V, Forjaz MJ. Disease burden morbidity assessment by self-report: Psychometric properties in older adults in Spain.Geriatrics & gerontology international Rodriguez-Blazquez C., Damian J., Andres-Prado M.J., Almazan-Isla J., Alcalde-Cabero E., Forjaz M.J. et al. Associations between chronic conditions, body functions, activity limitations and participation restrictions: A cross-sectional approach in Spanish non-clinical populations. BMJ Open. 2016;6(6). Damian J., Pastor-Barriuso R., Garcia Lopez F.J., de Pedro-Cuesta J. Urinary incontinence and mortality among older adults residing in care homes. Journal of Advanced Nursing Negron-Blanco L., De Pedro-Cuesta J., Almazan J., Rodriguez-Blazquez C., Franco E., Damian J. Prevalence of and factors associated with homebound status among adults in urban and rural Spanish populations. BMC Public Health. 2016;16(1). Alcalde-Cabero E., Almazan-Isla J., Garcia Lopez F.J., Ara-Callizo J.R., Avellanal F., Casasnovas C. et al. Guillain-Barré syndrome following the 2009 pandemic monovalent and seasonal trivalent influenza vaccination campaigns in Spain from 2009 to 2011: Outcomes from active surveillance by a neurologist network, and records from a country-wide hospital discharge database. BMC Neurology. 2016;16(1). Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Pino-Dominguez L., Navarro-Gil P., Gonzalez-Velez A.E., Prieto-Flores M.-E., Ayala A., Rojo-Perez F. et al. Self-perceived health status, gender, and work status. Journal of Women and Aging. 2016; 1-9. Royo-Bordonada M.A., Garcia Lopez F.J. Ethical considerations surrounding the response to Ebola: The Spanish experience. BMC Medical Ethics. 2016;17(1). Hopfner F., Nebel A., Lyons K.E., Troster A.I., Kuhlenbaumer G., Deuschl G. et al. Validation of the QUEST for German-speaking countries. International Journal of Neuroscience. 2016;126(2): CIBERNED Annual Report / 78

79 Program 1 Group: Jesús de Pedro Cuesta Royo-Bordonada M.A., Leon-Flandez K., Damian J., Bosqued-Estefania M.J., Moya-Geromini M.A., Lopez-Jurado L. The extent and nature of food advertising to children on Spanish television in 2012 using an international food-based coding system and the UK nutrient profiling model. Public Health. 2016;137: Royo-Bordonada M.T., Bosqued-Estefania M.J., Damian J., Lopez-Jurado L., Moya-Geromini M.T. Nutrition and health claims in products directed at children via television in Spain in Gaceta Sanitaria. 2016;30(3): Marventano S., Ayala A., Gonzalez N., Rodriguez-Blazquez C., Garcia-Gutierrez S., Forjaz M.J. Multimorbidity and functional status in institutionalized older adults. European Geriatric Medicine. 2016;7(1): Martinez Martin P, Rodriguez-Blazquez C. Evaluación de la enfermedad. En: Guía oficial de práctica clínica en la enfermedad de Parkinson. Coordinador: J.M. Arbelo González. Guías diagnósticas y terapéuticas de la Sociedad Española de Neurología. Luzán 5 S.S.: Madrid, 2016, pp: Meyer A., Hadinia A., Hatz F., Martinez-Martin P., Fuhr P., Stieglitz R.D. et al. German Translation and Validation of the Scale for Evaluation of Neuropsychiatric Disorders in Parkinson s disease (SEND-PD). Fortschritte der Neurologie Psychiatrie. 2016;84(7): de Pedro Cuesta J., Saiz Ruiz J., Roca M., Noguer I. Mental health and public health in Spain: Epidemiological surveillance and prevention. Psiquiatria Biologica. 2016;23(2): Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. Damian J. The aging of scientific material: Datagism. Epidemiology. 2016;27(2):e7-. Buetow SA, Martínez-Martín P, Hirsch MA, Okun MS. Beyond patient-centered care: personcentered care for Parkinson s disease. npj Parkinson s Disease. 2016; 2, Martínez-Martín P, Rojo-Abuin JM, Rodríguez-Violante M, Serrano-Dueñas M, Garretto N, Martínez-Castrillo JC. et al. Analysis of four scales for global severity evaluation in Parkinson s disease. npj Parkinson s Disease. 2016; 2, Ambrosio L, Portillo MC, Rodríguez-Blázquez C, Rodriguez-Violante M, Castrillo JCM, Arillo VC. et al. Living with chronic illness scale: international validation of a new self-report measure in Parkinson s disease. npj Parkinson s Disease. 2016; 2, Research projects Code: AGL REDT. Title: Red nacional de priones. Principal Investigator: José Antonio del Río. Colaboración CIBERNED: Yes. CIBERNED groups: G114, G509, G503, G609, G207. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: CINERNED 2014/02. Tile: Mecanismos epigenéticos implicados en la etiología y progresión de las demencias neurodegenerativas rápidamente progresivas. Principal Investigator: Miguel Calero. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G601. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: AMO-02-MD Title: GSK3 en la distrofia miotonica congénita de aparcición juvenil. Principal Investigator: Miguel Calero. CIBERNED groups: G509. Type: International. Funding agency: AMO PHARMA. Funding: Duration: CIBERNED Annual Report / 79

80 GROUP José Rodríguez Álvarez 406 Principal Investigator José Rodríguez Álvarez Research team José Aguilera Ávila Associate professor Ana María Candalija Iserte Judit Catala Solsona Lilian Enríquez Barreto Postdoctoral fellow Alfredo Jesús Miñano Postdoctoral fellow Laura Ortega Hernández Postdoctoral fellow Laura Rubio Ferrarons Technician Carlos Saura Antolín Associate professor Dolores Siedlecki Wullich Instituto de Neurociencias Universidad Autónoma de Barcelona Edificio M Campus de Bellaterra Cerdanyola del Vallés, Catalunya (Spain) Tel.: jose.rodriguez@uab.es 2016 CIBERNED Annual Report / 80

81 Summary 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 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). However, the role of CRTC1 and its regulatory mechanisms in hippocampal-dependent learning and memory encoding and loss, and specifically during neurodegeneration, are unknown. During the last year, our studies have revealed for the first time that CRTC1 is specifically activated in the hippocampus during associative memory encoding, and that disruption of CRTC1 activation underlies associative memory deficits and dendritic degeneration. Moreover, we have been exploring the underlying molecular mechanisms, related to glutamate AMPA receptors (AMPAR) regulation, involved in early learning and memory dysfunction associated to AD and to identify novel therapeutic targets and biomarkers for promoting prevention and diagnosis at MCI and early stages of the disease. In this context, we have identified a group of mirnas, related to AMPAR and other synaptic proteins regulation, that change significantly their expression levels in human samples of early AD. β-amyloid effect on synaptic activity could also be mediated by an alteration in the blood brain barrier, causing a misbalance in angineurins homeostasis. By using an in vitro model of blood-brain barrier we have observed that alteration in certain endothelial oxidases potentiate the segregation of β-amyloid and causes changes in BDNF and other angineurins levels, affection synaptic maturation. Finally, we have been also studying the relationship between mutation in the PS/γsecretase gene, the processing of the ephrin receptor EphA3 and the alteration of axonal growth and development. Keywords Alzheimer disease, memory, intracelular signaling, gene regulation, glutamate receptors, CRTC1, early synaptic dysfunction Publications Palafox-Sanchez V., Mendieta L., Ramirez-Garcia G., Candalija A., Aguilera J., Limon I.D. Effect of the C-terminal domain of the heavy chain of tetanus toxin on dyskinesia caused by levodopa in 6-hydroxydopamine-lesioned rats. Pharmacology Biochemistry and Behavior. 2016;145: Mendieta L., Granado N., Aguilera J., Tizabi Y., Moratalla R. Fragment C domain of tetanus toxin mitigates methamphetamine neurotoxicity and its motor consequences in mice. International Journal of Neuropsychopharmacology. 2016;19(8):1-11. Olivan S., Calvo A.C., Rando A., Herrando-Grabulosa M., Manzano R., Zaragoza P. et al. Neuroprotective effect of non-viral gene therapy treatment based on tetanus toxin C-fragment in a severe mouse model of spinal muscular atrophy. Frontiers in Molecular Neuroscience. 2016;9. Proyectos de investigación Code: 2015/01-1. Title: Terapia génica dirigida a neuregulinas para el tratamiento de enfermedades degenerativas de las motoneuronas. Principal Investigator: Xavier Navarro. CIBERNED s collaboration: Yes. CIBERNED groups: G607, G113, G406, G407. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 81

82 Code: SAF R. Title: Mecanismos moleculares implicados en la disfunción de la sinapsis glutamatergicas en fases tempranas de la enfermedad de Alzheimer. Principal Investigator: José Rodríguez Álvarez. CIBERNED groups: G406. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: 2014 SGR Title: Recerca Biomedica en Neurodegeneracio (REBINE). Principal Investigator: José Rodríguez Álvarez. CIBERNED groups:g 406. Type: CC.AA. Funding agency: AGAUR-Generalitat Catalunya. Funding: Duration: Code: A S. Title: Transcriptional mechanisms of memory loss in Alzheimer s disease. Principal Investigator: Carlos A. Saura. CIBERNED groups: G 406. Type: International. Funding agency: BrightFocus Foundation. Funding: Duration: Code: Marato TV Title: Searching new biomarkers and therapeutic targets related to cognitive deficits in early stages of Alzheimer s Disease: Role of AKAP79/150, CPT1C and SSAO/VAP- 1 in Ab-mediated AMPAR dysfunction. Principal Investigator: José Rodríguez Álvarez. CIBERNED groups: G406. Type: Private. Funding agency: Fundación La Maraton TV3. Funding: Duration: Code: SAF Title: Transcriptional mechanisms underlying memory loss in Alzheimer s disease transgenic mouse models (TRANSMECAD). Principal Investigator: Carlos A. Saura. CIBERNED groups: G 406. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: PhD dissertations Author: Arnaldo J. Parra Damas. Title: Role of the CREB/CRTC1-regulated gene transcription during hippocampaldependent memory in Alzheimer s disease mouse models. Date: 5 February Supervisor: Carlos A. Saura Antolín CIBERNED Annual Report / 82

83 GROUP Javier Sáez Valero 407 Principal Investigator Javier Sáez Valero Research team Inmaculada Cuchillo Ibáñez Postdoctoral fellow Inmaculada López Font Postdoctoral fellow Trinidad Mata Balaguer Postdoctoral fellow María Salud García Ayllón Postdoctoral fellow Jordi Alom Poveda Chair of Neurology department Aitana Sogorb Esteve Claudia P. Boix Instituto de Neurociencias de Alicante Universidad Miguel Hernández-CSIC Avenida Ramón y Cajal, s/n Sant Joan d Alacant (Spain) Tel.: Fax: j.saez@umh.es 2016 CIBERNED Annual Report / 83

84 Summary The aim of our research group is focus into Alzheimer s disease (AD) from a basic point of view, but also regarding translational benefits including therapy and diagnostic applications. In the basic research line we are focusing in understand whether Reelin signaling pathway is impaired in the brain of Alzheimer s patients, in spite of the increasing brain protein levels, and how the β-amyloid peptide compromises the biological activity of the Reelin glycoprotein (see the original research article: Cuchillo-Ibañez et al., Sci Rep. 2016;6:31646; and the review: Cuchillo- Ibañez et al., J Alzheimers Dis. 2016;52:403). We also extend our analysis of interaction between the enzyme acetylcholinesterase (AChE) and key AD proteins, β-amyloid and tau hyperphosphorylated, now focusing in alternative splice variants of AChE which may serve non-cholinergic functions unrelated with their capacity to hydrolyze acetylcholine. We first confirmed that a prominent pool of enzymatically inactive AChE protein exists in the AD brain, despite decrease in enzymatic activity. We also found an increase in the protein and transcript levels of the non-cholinergic readthrough AChE (AChE-R) variant in AD patients. We proposed that differential expression of AChE variants in AD may reflect changes in the pathophysiological role of AChE, independent of cholinergic impairment or its role in degrading acetylcholine, which can be of particular relevance since AChE is the target for the palliative therapy of AD (Campanari et al., J Alzheimers Dis. 2016;53:831). We have also extended our research in new cerebrospinal fluid (CSF) biomarkers for AD. We have been pioneers in demonstrating that Presenilin-1 (PS1), the active component of the intramembrane γ-secretase complex, can be detected as soluble heteromeric aggregates in CSF. Our new data from individuals with symptomatic and asymptomatic genetically determined AD, as well from subjects with Down syndrome (with β-amyloid over-expression), suggest that CSF-PS1 complexes may be useful as an early biomarker for AD, reflecting the pathology at asymptomatic state (Sogorb- Esteve et al., Mol Neurodegener. 2016;11:66). Moreover, we continue to explore the possibility that changes in glycosylation of proteins served to refine AD biomarkers. In a recent study we addressed the analysis that the regulation of the HNK-1 glycan results altered in AD. HNK-1 is a 3-sulfonated glucuronic acid glycoepitope, a marker of cell adhesion that participates in cell-cell and cell-extracellular matrix interactions and in neurite growth. We demonstrate an overall decrease in HNK-1 carrier glycoproteins extracted from the frontal cortex of AD subjects, as well in transgenic mice model of AD (Tg2576) and culture cells treated with the amyloidogenic Aβ42 peptide. These results provide evidence that cellular levels of HNK-1 carrier glycoforms are decreased in the brain of AD subjects, probably influenced by the β-amyloid protein. However, the levels of soluble HNK-1 glycoproteins detected in culture media were not affected by Aβ treatment, neither in CSF from AD patients (García-Ayllón et al. Mol Neurobiol. 2017;54:188; published online in 2016). Finally, in collaboration with other groups from CIBERNED we started to investigate molecular mechanism affected in Amyotrophic Lateral Sclerosis (ALS). In our first report is this disease we reviewed the neuromuscular junction impairment in ALS, with particular interest in reassessing the role of AChE in this disease. We have also started to characterize impaired Neuregulin 1 / ErbB4 signaling in ALS. Keywords Alzheimer, amyotrophic lateral sclerosis, β-amyloid, AChE, presenilin, P-tau, reelin, biomarker Publications Garcia-Ayllon M.-S., Botella-Lopez A., Cuchillo-Ibanez I., Rabano A., Andreasen N., Blennow K. et al. HNK-1 Carrier Glycoproteins Are Decreased in the Alzheimer s Disease Brain. Molecular Neurobiology. 2016; López-Riquelme N, Alom-Poveda J, Viciano-Morote N, Llinares-Ibor I, Tormo-Díaz C. Apolipoprotein E ε4 allele and malondialdehyde level are independent risk factors for Alzheimer s disease.sage open medicine. ; CIBERNED Annual Report / 84

85 Program 1 Group: Javier Sáez Valero Campanari ML, García-Ayllón MS, Ciura S, Sáez-Valero J, Kabashi E. Neuromuscular Junction Impairment in Amyotrophic Lateral Sclerosis: Reassessing the Role of Acetylcholinesterase. Frontiers in molecular neuroscience. 2016;9. Sogorb-Esteve A., Garcia-Ayllon M.-S., Fortea J., Sanchez-Valle R., Lleo A., Molinuevo J.-L. et al. Cerebrospinal fluid Presenilin-1 increases at asymptomatic stage in genetically determined Alzheimer s disease. Molecular Neurodegeneration. 2016;11(1):1-11. Campanari M.-L., Navarrete F., Ginsberg S.D., Manzanares J., Saez-Valero J., Garcia-Ayllon M.-S. Increased Expression of Readthrough Acetylcholinesterase Variants in the Brains of Alzheimer s Disease Patients. Journal of Alzheimer s Disease. 2016;53(3): Cuchillo-Ibanez I., Balmaceda V., Mata-Balaguer T., Lopez-Font I., Saez-Valero J. Reelin in Alzheimer s Disease, Increased Levels but Impaired Signaling: When More is Less. Journal of Alzheimer s Disease. 2016;52(2): Cuchillo-Ibanez I., Mata-Balaguer T., Balmaceda V., Arranz J.J., Nimpf J., Saez-Valero J. The β-amyloid peptide compromises Reelin signaling in Alzheimer s disease. Scientific Reports. 2016;6. Research projects Code: 2015/01-1. Title: Terapia génica dirigida a neuregulinas para el tratamiento de enfermedades degenerativas de las motoneuronas. Principal Investigator: Xavier Navarro. CIBERNED s collaboration: Yes. CIBERNED groups: G607, G113, G406, G407. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: UGP Title: Análisis de las variantes de AChE en cerebro control y con enfermedad de Alzheimer. Principal Investigator: María Salud García Ayllon. CIBERNED groups: G407. Type: CC.AA. Funding agency: Comunidad Valenciana. Funding: Duration: Code: PI14/ Title: Procesamiento por secretasas de la forma colinérgica de acetilcolinesterasa, su actividad transcripcional e implicación en la enfermedad de Alzheimer. Principal Investigator: María Salud García Ayllon. CIBERNED groups: G407. Other CIBER s collaborarion: No. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: ,99. Duration: Code: SAF CIN. Title: 1st IN PhD Student & Postdoc Meeting Building Neuroscience: The future of a multidisciplinary field. Principal Investigator: Javier Sáez Valero. CIBERNED groups: G407. Type: National. Funding agency: Ministerio de Educación, Cultura y Deporte. Funding: Duration: Code: PI15/ Title: Interacciones de las vias Reelina/ApoE y ligandos alternativos con el β-amiloide, su disfunción en la enfermedad de Alzheimer. Principal Investigator: Javier Sáez Valero. CIBERNED groups: G407. Othere CIBER s collaboration: No. Type: National. Funding agency: Instituto de Saud Carlos III. Funding: Duration: CIBERNED Annual Report / 85

86 GROUP Eduardo Soriano García 408 Principal Investigator Eduardo Soriano García Full professor Research team Marta Pascual Sánchez Researcher Jesús Mariano Ureña Bares Researcher Ferran Burgaya Márquez Researcher Ashraf Muhaisen Postdoctoral fellow Serena Mirra Postdoctoral fellow Daniela Rossi Postdoctoral fellow Antoni Parcerisas Mosqueda Cristina Roselló Busquets Marc Hernaiz Llorens Fausto Alexánder Ulloa Darquea Researcher Tiziana Cotrufo Postdoctoral fellow Lluís Pujadas Puigdomènech Postdoctoral fellow Yasmina Manso Sanz Postdoctoral fellow Mónica Pardo Muñoz Postdoctoral fellow Jonatan Dorca Arévalo Postdoctoral fellow Universidad de Barcelona Facultad de Biología Departamento de Biología Celular Avenida Diagonal 643, Edificio Prevosti 1r piso Tel.: Fax: Irene Lobón García Alba del Valle Vilchez Acosta Marco Solís Benites Research assistante Núria Masachs Janoher Pablo Barrecheguren Manero 2016 CIBERNED Annual Report / 86

87 Summary In Alzheimer s disease less than 1.5% of cases are hereditary (FAD), while the causes of lateonset sporadic Alzheimer s disease (SAD) remain largely unknown. Several genetic factors (identified in GWAS) increase the risk of developing this disease. Throughout this year we have focused mainly on two hypotheses regarding the pathogenesis of AD and potential therapeutic strategies: 1) Reelin is an extracellular protein crucial for brain development, which is also expressed in the adult brain. Recent studies support the relationship between Reelin and AD and we have hypothesized that by regulating metabolism Aß42, phosphorylation of Tau and synaptic plasticity in adults, the Reelin cascade may be an important regulator of brain functions in adults, whose deregulation may be related to the pathology of AD, suggesting that Reelin may be protective against AD. We recently tested this hypothesis by overexpression of Reelin (Reelin-OE mice) in a model of AD (J20). The Reelin-OE / J20 mice showed decreased plaque burden and the rescue of synaptic and cognitive deficits, suggesting that Reelin protects from AD pathology. We have studied: If the Reelin improves AD in mice, once the disease has developed, and in mice that exhibit AD pathology related to Tau (Tau-GSK3 WLW -OE and mice). Whether a downregulation of Reelin specifically in the adult (using conditional KO mice, FlReelin / AD mice) accelerates AD pathology. Started to understand the physiological and molecular mechanisms by which affects Reelin AD pathology, including 2) genetic and molecular Reelin-dependent networks. Some human diseases, including cancer, are caused by somatic mutations that alter the function of specific cell populations. Through the complete sequencing of the exome we have recently tested the hypothesis that variations and somatic mutations (referenced here as Single Nucleotide Variations-SNVS) are present in the brain of SAD. We have found a remarkable number of specific SNVS in SAD, which were not detected in the blood of the same donors. SNVS specific loci with recurrent SNVS were common to several patients and were not found in the brains of control donors. These results reveal that the blood and brain have different genomic DNA variations and suggest that somatic genome remodeling of the brain may contribute to the pathogenesis of SAD. In this year we have tried to understand more deeply the signing of somatic mutations in brain SAD: Determining somatic gene signature in SAD, including INFDELS, variations on chromosome X and copy number variations (CNVs). Integrating these data in defined molecular pathways through systems biology. We have begun to understand how specific somatic genetic events SAD may contribute to the pathogenesis of the disease. We believe these data reveal important new molecular and genetic information to understand the mechanisms that trigger the SAD and to open new therapeutic avenues. Keywords Reelin, synapses, Alzheimer disease, somatic mutations Publications Soler H., Dorca-Arevalo J., Gonzalez M., Rubio S.E., Avila J., Soriano E. et al. The GABAergic septohippocampal connection is impaired in a mouse model of tauopathy. Neurobiology of Aging. 2017;49: Epub CIBERNED Annual Report / 87

88 Gomez-Ramos A., Picher A.J., Garcia E., Garrido P., Hernandez F., Soriano E. et al. Validation of Suspected Somatic Single Nucleotide Variations in the Brain of Alzheimer s Disease Patients. Journal of Alzheimer s Disease. 2017;56(3): Epub Barrecheguren P.J., Ros O., Cotrufo T., Kunz B., Soriano E., Ulloa F. et al. SNARE proteins play a role in motor axon guidance in vertebrates and invertebrates. Developmental Neurobiology. 2017;. Epub Labouesse M.A., Lassalle O., Richetto J., Iafrati J., Weber-Stadlbauer U., Notter T. et al. Hypervulnerability of the adolescent prefrontal cortex to nutritional stress via reelin deficiency. Molecular Psychiatry Bosch C, Masachs N, Exposito-Alonso D, Martínez A, Teixeira CM, Fernaud I et al. Reelin Regulates the Maturation of Dendritic Spines, Synaptogenesis and Glial Ensheathment of Newborn Granule Cells.Cerebral cortex (New York, N.Y. : 1991) Sanchez-Huertas C., Freixo F., Viais R., Lacasa C., Soriano E., Luders J. Non-centrosomal nucleation mediated by augmin organizes microtubules in post-mitotic neurons and controls axonal microtubule polarity. Nature Communications. 2016;7. Bosch C., Muhaisen A., Pujadas L., Soriano E., Martinez A. Reelin exerts structural, biochemical and transcriptional regulation over presynaptic and postsynaptic elements in the adult hippocampus. Frontiers in Cellular Neuroscience. 2016;10(MAY). Bosch C, Martínez A, Masachs N, Teixeira CM, Fernaud I, Ulloa F et al. Corrigendum: FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFPlabeled adult-generated neurons.frontiers in neuroanatomy. 2016;10. Martinez-Marmol R., Barneda-Zahonero B., Soto D., Andres R.M., Coccia E., Gasull X. et al. FAIM-L regulation of XIAP degradation modulates Synaptic Long-Term Depression and Axon Degeneration. Scientific Reports. 2016;6. Mirra S., Ulloa F., Gutierrez-Vallejo I., Marti E., Soriano E. Function of Armcx3 and Armc10/SVH genes in the regulation of progenitor proliferation and neural differentiation in the chicken spinal cord. Frontiers in Cellular Neuroscience. 2016;10(MAR2016). Masdeu M.D.M., Armendariz B.G., Soriano E., Urena J.M., Burgaya F. New partners and phosphorylation sites of focal adhesion kinase identified by mass spectrometry. Biochimica et Biophysica Acta - General Subjects. 2016;1860(7): Research projects Code: PI2016/04. Title: The ALS CIBERNED Challenge: accelerating new drug discovery. Principal Investigator: Adolfo López De Munain Arregui. CIBERNED s collaboration: Yes. CIBERNED groups: G609, G303, G503, G408. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: 613/C/2013. Title: Lipotoxicity, hepatic steatosis and hepatocarcinoma: Role of the ARMC10/ ARMCX family of mitochondrial proteins. Principal Investigator: Eduardo Soriano. CIBERNED groups: G408. Other CIBER s collaboration: Yes (CIBER Fisiopatologia de la Obesidad y Nutricion). Type: Private. Funding agency: Fundació la Marato de TV3. Funding: Duration: Code: 2014 SGR Title: Neurobiologia del Desenvolupament i Regeneració Neuronal (SGR ). Principal Investigator: Eduardo Soriano García. CIBERNED groups: G408. Type: CC.AA. Funding agency: AGAUR-Generalitat de Catalunya Funding: Duration: CIBERNED Annual Report / 88

89 Program 1 Group: Eduardo Soriano García Code: SAF R. Title: Nuevas aproximaciones a la patogénesis de la enfermedad de Alzheimer. Principal Investigator: Eduardo Soriano Garcia. CIBERNED groups: G408. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: Title: The role of reelin at the crossroads of alzheimer's disease mechanisms: Tauopathy, amyloid toxicity and transmissibility. Principal Investigator: Lluis Pujadas Puigdomenech. CIBERNED groups: G408. Type: Private. Funding agency: Fundació la Marato de TV3. Funding: ,5. Duration: CIBERNED Annual Report / 89

90 GROUP Ignacio Torres Alemán 409 Principal Investigator Ignacio Torres Alemán Research team Ana M. Fernández Researcher Miguel García Researcher Dolores Guinea Researcher Víctor Munive Researcher Andrea Santi Researcher Angélica Stein Researcher James Knight Researcher Paloma Pérez Domper Researcher Jonathan Zegarra Researcher Instituto Cajal Avenida Dr Arce, Madrid (Spain) Tel.: Fax: CIBERNED Annual Report / 90

91 Summary Our group is working on the role played by insulin peptides in brain homeostasis and its impact on brain pathologies. During 2016 we continued studying the mechanisms underlying control of brain glucose metabolism by IGF-I and insulin and their significance in Alzheimer s disease. Additional studies include the differences in mood homeostasis along aging in female mice. Current evidence suggests that the high incidence of mood co-morbidities in Alzheimer patients may be related to changes that we observed are taking place relatively early at middle age stages. Along the same line, we have almost completed our mechanistic studies pointing to a role of the relation of the glucocorticoid receptor chaperone FKBP5 with IGF-I that may translate to human biology. In addition, while analyzing mechanisms of entrance of serum IGF-I into the brain, we have gathered evidence of a potentially significant therapeutic action of IGF-I in brain injury, probably including stroke. Finally, we are looking into mechanisms connecting type 2 diabetes and sporadic Alzheimer disease, two age-related conditions. Keywords Insulin factors, neuronal plasticity, and degenerative diseases, therapeutic targets in neurodegeneration, the aging brain and cognition, blood brain barrier, mood homeostasis Publications Fernandez A.M., Hernandez-Garzon E., Perez-Domper P., Perez-Alvarez A., Mederos S., Matsui T. et al. Insulin regulates astrocytic glucose handling through cooperation with IGF-I. Diabetes. 2017;66(1): Epub Nishijima T, Torres-Aleman I, Soya H. Exercise and cerebrovascular plasticity. In: Kazuto Masamoto, Hajime Hirase and Katsuya Yamada, editors, Progress in Brain Research, Vol. 225, Amsterdam: Elsevier, 2016, pp ISBN: Shima T., Matsui T., Jesmin S., Okamoto M., Soya M., Inoue K. et al. Moderate exercise ameliorates dysregulated hippocampal glycometabolism and memory function in a rat model of type 2 diabetes. Diabetologia. 2016; Garcia-Caceres C., Quarta C., Varela L., Gao Y., Gruber T., Legutko B. et al. Astrocytic Insulin Signaling Couples Brain Glucose Uptake with Nutrient Availability. Cell. 2016;166(4): Diez H., Benitez M.J., Fernandez S., Torres-Aleman I., Garrido J.J., Wandosell F. Class I PI3-kinase or Akt inhibition do not impair axonal polarization, but slow down axonal elongation. Biochimica et Biophysica Acta - Molecular Cell Research. 2016;1863(11): Trueba-Saiz Á, Fernandez AM, Nishijima T, Mecha M, Santi A, Munive V et al. Circulating Insulinlike Growth Factor I Regulates Its Receptor in the Brain of Male Mice.Endocrinology Hernandez-Garzon E., Fernandez A.M., Perez-Alvarez A., Genis L., Bascunana P., Fernandez de la Rosa R. et al. The insulin-like growth factor I receptor regulates glucose transport by astrocytes. GLIA Munive V., Santi A., Torres-Aleman I. A Concerted Action of Estradiol and Insulin Like Growth Factor i Underlies Sex Differences in Mood Regulation by Exercise. Scientific Reports. 2016;6. Fernandez A.M., Hervas R., Dominguez-Fraile M., Garrido V.N., Gomez-Gutierrez P., Vega M. et al. Blockade of the Interaction of Calcineurin with FOXO in Astrocytes Protects Against Amyloidβ-Induced Neuronal Death. Journal of Alzheimer s Disease. 2016;52(4): Trueba-Saiz A, Aleman I. Insulin-like peptides signaling in Alzheimer s disease: on the road to alternative therapeutics. Current Opinion in Behavioral Sciences. 2016; 9: CIBERNED Annual Report / 91

92 Research projects Code: S2010_BMD. Title: Redes de senalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas. Principal Investigator: José González Castaño. CIBERNED s collaboration: Yes. CIBERNED groups: G111, G401, G104, G307, G409, G412. Other CIBER s collaboration: Yes (CIBERER). Type: CC.AA. Funding agency: Comunidad de Madrid. Funding: Duration: Code: PI2016/01. Title: Alteraciones del metabolismo glucolipídico y desarrollo de la demencia de Alzheimer. Principal Investigator: Ignacio Torres Alemán. CIBERNED s collaboration: Yes. CIBERNED groups: G409, G402, G511, G502, G412. Type: Intramurales Funding agency: CIBERNED. Funding: Duration: Code: CSIC13-4E Title: Potenciacóon de los servicios científicos del Instituto Cajal. Principal Investigator: Ignacio Torres Alemán. CIBERNED groups: G409. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 92

93 GROUP Ramón Trullás Oliva 410 Principal Investigator Ramón Trullás Oliva CSIC research professor Research team Anna Colell Riera Research scientist Petar Podlesniy Research scientist Nicole Mahy Professor of Biochemistry Manuel Rodríguez Allué Assistant professor Nuria Serra Technician Margalida Puigròs Predoctoral student Unidad de Neurobiología IIBB-CSIC IDIBAPS Calle Rosselló 161, 7ª Planta, Laboratorio Barcelona (Spain) Tel.: Fax: CIBERNED Annual Report / 93

94 Summary Throughout the year 2016 we have studied the processes involved in the regulation of mitochondrial DNA replication, transcription and release (mtdna) in cellular models of neurodegenerative diseases and in fibroblasts from patients with Parkinson s disease. The results obtained confirm that a decrease in the concentration of mtdna in cerebrospinal fluid (CSF) precedes the appearance of clinical symptoms of both sporadic and familial Alzheimer s disease. In addition, we investigated whether mtdna concentration in CSF can distinguish Alzheimer s disease from other types of rapidly progressing dementias, because it is necessary to identify urgently biomarkers that are capable of differentiating different dementia subtypes. The results show that a decrease greater than 50% in the mtdna concentration in CSF distinguishes Alzheimer s dementia from other subtypes of dementia, including that associated with Creutzfeldt-Jakob disease. These results indicate that mtdna content in the CSF may allow the differential diagnosis of Alzheimer s disease. In addition, results obtained in patients with Creutzfeldt-Jakob disease indicate that mtdna in the CSF does not come from neuronal damage. On the other hand, we studied the mtdna concentration in the CSF of patients with idiopathic Parkinson s disease, and in patients bearing the G2019S mutation in the LRRK2 gene, that causes Parkinson s disease in most carriers. The results obtained show that patients with the G2019S mutation in the LRRK2 gene that also manifest Parkinson s disease have a higher concentration of mtdna in the CSF compared to both non-manifesting mutation carriers and patients Idiopathic Parkinson s disease. In addition, the results confirm that there is a low concentration of alpha-synuclein in the CSF in idiopathic Parkinson s disease, but not in Parkinson s disease caused by the mutation of the LRRK2 gene. Overall, our results demonstrate that mtdna concentration in the CSF distinguishes idiopathic Parkinson s disease from that caused by the mutation in the LRRK2 gene, and indicate that the neurodegenerative process occurring in these two types of Parkinson s disease is caused by different biochemical mechanisms. Future studies in our laboratory will focus on identifying the mechanism of mtdna release and the relationship of its release to the neurodegenerative process. 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 Podlesniy P, Llorens F, Golanska E, Sikorska B, Liberski P, Zerr I et al. Mitochondrial DNA differentiates Alzheimer from Creutzfeldt-Jakob disease.alzheimer s & dementia : the journal of the Alzheimer s Association Espinosa-Parrilla J, Pugliese M, Mahy M, Rodríguez MJ. Neuroprotection: A New Therapeutic Approach of Relapsing Remitting Multiple Sclerosis. In: Trending Topics in Multiple Sclerosis. Gonzalez-Quevedo A, editors. InTech, 2016, pp ISBN Martinez-Moreno M., Batlle M., Ortega F.J., Gimeno-Bayon J., Andrade C., Mahy N. et al. Diazoxide enhances excitotoxicity-induced neurogenesis and attenuates neurodegeneration in the rat non-neurogenic hippocampus. Neuroscience. 2016;333: Podlesniy P., Vilas D., Taylor P., Shaw L.M., Tolosa E., Trullas R. Mitochondrial DNA in CSF distinguishes LRRK2 from idiopathic Parkinson s disease. Neurobiology of Disease. 2016;94: CIBERNED Annual Report / 94

95 Program 1 Group: Ramón Trullás Oliva Research projects Code: PI2016/06. Title: Identificación de vías fisiopatológicas y biomarcadores candidatos en la fase prediagnóstica de la enfermedad de Parkinson. Principal Investigator: Miquel Vila Bover. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G601, G207, G410. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: LRRK2 Resource RFA Title: Alpha-synuclein and other biomarkers in biological samples of LRKK2 PD. Principal Investigator: Eduardo Tolosa. CIBERNED s collaboration: Yes. CIBERNED groups: G410, G207. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: Code: PI Title: Mitochondrial DNA copy number in LRRK2 fibroblasts. Principal Investigator: Eduardo Tolosa. CIBERNED s collaboration: Yes. CIBERNED groups: G410, G207. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: Code: SAF R. Title: Mecanismos de regulación del número de copias de ADN mitocondrial para el tratamiento del alzhéimer. Principal Investigator: Ramón Trullás. CIBERNED groups: G410. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 95

96 GROUP Javier Vitorica Ferrández 411 Principal Investigator Javier Vitorica Ferrández Research team Marisa Vizuete Chacón Full professor Victoria Navarro Garrido Fellow MªVirtudes Sánchez Mica Fellow Clara Muñoz Castro Fellow Sebastián Jiménez Muñoz Research staff Mª Luisa García-Calvo Technician Dpto. Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Sevilla Instituto de Biomedicina de Sevilla, Hospital Universitario Virgen del Rocío Sevilla (Spain) Tel.: Fax: vitorica@us.es 2016 CIBERNED Annual Report / 96

97 Summary In 2016 our group, in collaboration with the group of Dr. A. Gutierrez, University of Málaga, has completed the characterization of the microglial response in hippocampus from samples of patients with various Braak stages, from Braak 0 to Braak V-VI. The latter group corresponds to Alzheimer disease dementia. Our data demonstrated the existence of limited microglial activation within Alzheimer s patients. This limited activation could be explained by the relatively low accumulation of Abeta plaques in this particular region. Furthermore, the limited activation clearly contrasts with what was classically observed in transgenic models based on Abeta production. On the other hand, our results also demonstrated the existence of a microglial degenerative process in the cohort of Alzheimer patients. This microglial degenerative process has been analyzed both by specific microglial markers and by morphometric and stereological studies analysis in the different patients. Finally in this regard, we have also analyzed the possible factors involved in microglial degeneration. The data obtained demonstrate the presence of the phosphorylated tau protein within the soluble fractions of Alzheimer s samples. In addition, in vitro studies using flow cytometry and phagocytosis assays demonstrated that these phosphorylated forms of tau protein, both extra- and/or intracellular, were highly toxic to microglial cells. We are currently extending this study to other brain areas, with higher levels of Abeta deposition, such as frontal cortex. On the other hand, we have continued investigating the response of astrocytes in this same cohort of human samples. The results obtained to date showed that, unlike the microglial response, astrocytes in Alzheimer s patients were active and not affected by the degenerative process observed in microglia. However, we do not know yet the function of the astroglial activation of the evolution of the pathology. This objective of our current work is included in an intramural project CIBERNED (I.P. Joan Comella) and is also performed in collaboration with Dr. E. Galea (Maratón de TV3). Finally, we are also determined the role of microglial activation in the evolution of Alzheimer s disease using tau, APP or PS1xAPP models. In particular, tau models will be of great help in investigating both the molecular mechanisms involved in microglial death and the formation of soluble forms of phosphorylated tau. On the other hand, Abeta-based models will be used to manipulate the microglial response, inhibiting or increasing the response, either pharmacologically (immunosuppression) or genetically (IL-4-KO; Galectin3 KO; CSF1R- KO). After the microglial manipulation the the pathology (Abeta accumulation, tau phosphorylation, neurodegeneration) of the transgenic models will be analyzed. Keywords Alzheimer s, degeneration, inflammation, Abeta, tau, oligomers, soluble, transgenic models, neuropathology Publications Sanchez-Mejias E., Navarro V., Jimenez S., Sanchez-Mico M., Sanchez-Varo R., Nunez-Diaz C. et al. Soluble phospho-tau from Alzheimer s disease hippocampus drives microglial degeneration. Acta Neuropathologica. 2016; Hernández-Rodríguez M, Correa-Basurto J, Gutiérrez A, Vitorica J, Rosales-Hernández MC. Asp32 and Asp228 determine the selective inhibition of BACE1 as shown by docking and molecular dynamics simulations.european journal of medicinal chemistry. 2016;124. Fernandez A.M., Hervas R., Dominguez-Fraile M., Garrido V.N., Gomez-Gutierrez P., Vega M. et al. Blockade of the Interaction of Calcineurin with FOXO in Astrocytes Protects Against Amyloidβ-Induced Neuronal Death. Journal of Alzheimer s Disease. 2016;52(4): CIBERNED Annual Report / 97

98 Research projects Code: PI2015-2/02. Title: Potencial patológico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal Investigator: Joan X. Comella Carnice. CIBERNED s collaboration: Yes. CIBERNED groups: G413, G415, G204, G108, G411. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: Title: Deciphering the link between astrocyte reactivity and neuronal damage in Alzheimer s disease. Principal Investigator: Elena Galea. CIBERNED s collaboration: Yes. CIBERNED groups: G411, G413, G415. Type: Private. Funding agency: La Marató de TV3. Funding: Duration: Code: CTS Title: Oligomerización y toxicidad de los peptidos de Abeta: búsqueda de nuevas dianas de interes terapéutico en la enfermedad de Alzheimer. Principal Investigator: Javier Vitorica. CIBERNED s collaboration: Yes. CIBERNED groups: G411, G415. Type: CCAA. Funding agency: Junta de Andalucía. Funding: ,5. Duration: Code: FIS PI15/ Title: Evaluando la disfunción microglial y astroglial como base del proceso neurodegenerativo y la demencia en la enfermedad de Alzheimer: nuevas aproximaciones terapéuticas. Principal Investigator: Antonia Gutiérrez Pérez. CIBERNED s collaboration: Yes. CIBERNED groups: G415, G411. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: PhD dissertations Author: Sebastián Jiménez Muñoz. Title: Efectos de la acumulación del péptido Beta Amiloide en tejido cerebral de ratones PS1xAPP modelos de la enfermedad de Alzheimer. Date: 15 January Supervisor: Francisco Javier Vitorica Ferrández CIBERNED Annual Report / 98

99 GROUP Francisco Wandosell Jurado 412 Principal Investigator Francisco Wandosell Jurado Staff scientist Research team Inés M. Antón Staff scientist, researcher María José Benítez Moreno Associate professor María José Pérez Álvarez Assistant professor, UAM Juan José Garrido Jurado CSIC staff scientist, researcher Lara Ordóñez Gutiérrez Postdoctoral fellow Irene Benito Centro de Biología Molecular Severo Ochoa CSIC-UAM Universidad Autónoma de Madrid Calle Nicolás Cabrera nº1, Cantoblancon Madrid (Spain) Tel.: /45 91 Fax: CIBERNED Annual Report / 99

100 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 (IP s-f Wandosell (CBM), I. Anton (CNB) and J.J. Garrido, Cajal Inst.). 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 speciespecific, 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 some key points of these signalling. Our initial studies were focused on axonal differentiation and polarization, we defined the role of PI3K class I, GSK3 and Akt in the neuronal morphogenesis using pharmacological and shrna approaches. We reported that this via is modified after Ischemia and that the neuroprotective role of neurosteroids, such as estradiol, is due to the activation of PI3K-Akt pathway, at least in part (CoIPs: Dr. J.J.Garrido & F Wandosell). Second, we are analysing the role of the Akt downstream element, mtorc1. This protein complex controls general aspects of protein synthesis and autophagy; whereas its dysfunction has been considering a key factor of beta amyloid generation and/ or degradation in Alzheimer disease. (IPs: F. Wandosell). Third, we are analysing some Aktdownstream elements (such as FoxO, Bim, β-catenin, YAP/TAZ), and our data indicated that they are responsible of the cell division of cancer-stem cells and the maintenance of its phenotype. Our work is now focusing on the conversion from astrocyte-astrocytoma-glioma trying to define the role Akt-downtream elements and how WIP regulates actin cytoskeleton and glioma proliferation (CoIPs: Dr. 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, estrogens, Alzheimer s disease, neuritogenesis, axonal polarity, glia growth, astrocyte / glioma conversion, cytoskeleton, actin Publications Ordonez-Gutierrez L., Posado-Fernandez A., Ahmadvand D., Lettiero B., Wu L., Anton M. et al. ImmunoPEGliposome-mediated reduction of blood and brain amyloid levels in a mouse model of Alzheimer s disease is restricted to aged animals. Biomaterials. 2017;112: Epub Knafo S., Sanchez-Puelles C., Palomer E., Delgado I., Draffin J.E., Mingo J. et al. PTEN recruitment controls synaptic and cognitive function in Alzheimer s models. Nature Neuroscience Rodriguez-Feo J.A., Gallego-Delgado J., Puerto M., Wandosell F., Osende J. Reticulon-4B/Nogo-B acts as a molecular linker between microtubules and actin cytoskeleton in vascular smooth muscle cells. Biochimica et Biophysica Acta - Molecular Cell Research. 2016;1863(8): Mancini S, Minniti S, Gregori M, Sancini G, Cagnotto A, Couraud PO et al. The hunt for brain Aβ oligomers by peripherally circulating multi-functional nanoparticles: Potential therapeutic approach for Alzheimer disease.nanomedicine : nanotechnology, biology, and medicine. 2016;12(1). Mateos L., Perez-Alvarez M.J., Wandosell F. Angiotensin II type-2 receptor stimulation induces neuronal VEGF synthesis after cerebral ischemia. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2016;1862(7): CIBERNED Annual Report / 100

101 Program 1 Group: Francisco Wandosell Jurado Ordonez-Gutierrez L., Fernandez-Perez I., Herrera J.L., Anton M., Benito-Cuesta I., Wandosell F. AβPP/PS1 Transgenic Mice Show Sex Differences in the Cerebellum Associated with Aging. Journal of Alzheimer s Disease. 2016;54(2): Diez H., Benitez M.J., Fernandez S., Torres-Aleman I., Garrido J.J., Wandosell F. Class I PI3-kinase or Akt inhibition do not impair axonal polarization, but slow down axonal elongation. Biochimica et Biophysica Acta - Molecular Cell Research. 2016;1863(11): Gargini R., Garcia-Escudero V., Izquierdo M., Wandosell F. Oncogene-mediated tumor transformation sensitizes cells to autophagy induction. Oncology Reports. 2016;35(6): Cabrera J.R., Viejo-Borbolla A., Alcami A., Wandosell F. Secreted herpes simplex virus-2 glycoprotein G alters thermal pain sensitivity by modifying NGF effects on TRPV1. Journal of Neuroinflammation. 2016;13(1). Perez-Alvarez M.J., Wandosell F. Stroke and neuroinflamation: Role of sexual hormones. Current Pharmaceutical Design. 2016;22(10): Gargini R, Escoll M, García E, García-Escudero R, Wandosell F, Antón IM. WIP Drives Tumor Progression through YAP/TAZ-Dependent Autonomous Cell Growth.Cell reports. 2016;17(8). Research projects Code: PI2016/01. Title: Alteraciones del metabolismo glucolipídico y desarrollo de la demencia de Alzheimer. Principal Investigator: Ignacio Torres Aleman. CIBERNED s collaboration: Yes. CIBERNED groups: G409; G402; G511; G502; G412. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: SAF R. Title: Análisis de la senalización mediada por akt en neurodegeneración y en proliferación, migración/invasión celulares. Principal Investigator: Francisco Wandosell; Ines M. Antón Gutiérrez. CIBERNED groups: G412. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: OC Title: European Network of multidisciplinary research and translation of autophagy knowledge. Principal Investigator: Caty Casas. CIBERNED groups: G412. Type: European. Funding agency: Comision Europea. Funding: N.D. Duration: CIBERNED Annual Report / 101

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103 Program 2 Parkinson s disease and other neurodegenerative movement disorders Alberch Vie, Jordi Canela Campos, Enric Isidre Ceña Callejo, Valentín Cuadrado Pastor, Antonio Fariñas Gómez, Isabel Fernández Chacón, Rafael Fernández Ruiz, Javier Fuentes Rodríguez, José Manuel García Verdugo, José Manuel González Castaño, José Guzmán Pastor, Manuel Iglesias Vacas, Teresa Infante Ceberio Jon Kulisevsky Bojarski, Jaime Labandeira García, José Luis Lanciego Pérez, José Luis López Barneo, José López de Munain Arregui, Adolfo Lucas Lozano, José Javier Moratalla Villalba, Rosario Muñoz Cánoves, Pura Naranjo Orovio, José Ramón Navarro Acebes, Xavier Obeso Inchausti, José Ángel Pérez Castillo, Ana María Pérez Tur, Jordi del Río Fernández, José Antonio Rodríguez Díaz, Manuel Tolosa Sarró, Eduardo Vicario Abejón, Carlos Vila Bover, Miquel...218

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105 Program 2: Parkinson s disease and other neurodegenerative movement disorders This Program 2 brings together 31 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 Parkinson s disease (PD) is mainly 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. It is estimated to affect around 70,000 people in Spain, a figure that is expected to be increasing due to the progressive aging of the population. Although preventing and correcting DA deficit are still important goals, they cannot be considered the ultimate challenge in PD nowadays. Within this area, it is considered of vital importance to make progress on defining the following issues: 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. In this way, it is sought to achieve a true translational research nature, whose main objectives are the disease and the patient. 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. Picture: Ana María Pérez Castillo (110), Instituto de Investigaciones Biomédicas CSIC-UAM, Madrid 2016 CIBERNED Annual Report / 105

106 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. HD 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 production, etc. Some of these products offer encouraging results in experimental models of the disease but, unfortunately, not 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. Research Line 3: Neurodegenerative motor disorders 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, 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. 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 CIBERNED Annual Report / 106

107 Principal investigator Institution Alberch Vie, Jordi Canela Campos, Enric Isidre Ceña Callejo, Valentín Cuadrado Pastor, Antonio Del Río Fernández, José Antonio Fariñas Gómez, Isabel Fernández Chacón, Rafael Fernández Ruiz, Javier Fuentes Rodríguez, José Manuel García Verdugo, José Manuel González Castaño, José Guzmán Pastor, Manuel Iglesias Vacas, Teresa Infante Ceberio, Jon Kulisevsky Bojarski, Jaime Labandeira García, José Luis Lanciego Pérez, José Luis López Barneo, José López de Munain Arregui, Adolfo Lucas Lozano, José Javier Moratalla Villalba, Rosario Muñoz Cánoves, Pura Naranjo Orovio, José Ramón Navarro Acebes, Xavier Obeso Inchausti, José Ángel Pérez Castillo, Ana María Pérez Tur, Jordi Rodríguez Díaz, Manuel Tolosa Sarró, Eduardo Vicario Abejón, Carlos Vila Bover, Miquel University of Barcelona University of Barcelona University of Castilla La Mancha, Albacete Autonomous University of Madrid Catalonian Institute of Bioengineering, Barcelona University of Valencia University of Seville Complutense University of Madrid University of Extremadura, Cáceres Cavanilles Institute, University of Valencia Autonomous University of Madrid Complutense University of Madrid Biomedic al Research Institute CSIC-UAM, Madrid Instituto de Investigación Marqués de Valdecilla, Santander Santa Creu i Sant Pau Hospital, Barcelona University of Santiago de Compostela Center for Applied Medical Research, Univ. of Navarra, Pamplona Virgen del Rocío University Hospital, Universidad de Sevilla Biodonostia Research Institute, San Sebastian Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid Cajal Institute CSIC, Madrid Pompeu Fabra University, ICREA, Barcelona National center of Biotechnology CSIC, Madrid Autonomous University of Barcelona Hospitales de Madrid Foundation Biomedical Research Institutes CSIC-UAM, Madrid Biomedicine Institute CSIC, Valencia University of La Laguna, Tenerife Clinic Hospital, Barcelona Cajal Institute CSIC, Madrid Vall d Hebron University Hospital, Barcelona Program 2 is coordinated by Drs. José J. Lucas (Center for Molecular Biology, CSIC, Madrid), Eduardo Tolosa (Clinic Hospital, Barcelona), Adolfo López de Munain Arregui (Biodonostia Research Institute, San Sebastián), Rafael Fernández Chacón (University of Seville) y Teresa Iglesias Vacas (Biomedical Research Institute CSIC-UAM, Madrid) CIBERNED Annual Report / 107

108 GROUP Jordi Alberch Vié 301 Principal Investigator Jordi Alberch Vié Full professor Research team Josep M Canals Coll Associate professor Silvia Ginés Padrós Associate professor Esther Pérez Navarro Associate professor Glòria Blázquez Postdoctoral researcher Verónica Brito Postdoctoral researcher Raquel Martín Ibáñez Postdoctoral researcher Mercè Massana Postdoctoral researcher Andrés Míguez González Postdoctoral researcher Ana Cristina Saavedra Postdoctoral researcher Phil Sanders Postdoctoral researcher Marco Straccia Postdoctoral researcher Xavier Xifró Collsamata Assistant professor Rafael Alcalá Vida Elena Álvarez Periel Marta Cherubini Andrea Comella Bolla Jordi Creus Muncunill Sara Fernández García Marta García Forn Alfonso Gerardo García Díaz Barriga Inés Guardia Pena Laura López Molina Núria Suelves Caballol Laura Vidal Sancho Georgina Bombau Martínez Technician Ester González Guerrero Technician Cristina Herranz Sotoca Technician Ana López Alonso Technician Mª Teresa Muñoz Technician Unai Perpiñá Martín Technician Departament de Biologia Cellular, Inmunologia i Neurociències Facultat de Medicina, Universitat de Barcelona Institut d Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) Tel.: / Fax: alberch@ub.edu 2016 CIBERNED Annual Report / 108

109 Summary During 2016, we continue our studies to identify new intracellular targets to develop therapeutical approaches for Huntington s disease (HD) and other neurological disorders. We show that targeting CAG repeat RNAs reduces HD phenotype independently of huntingtin levels, indicating the relevance of expanded CAG RNA to HD pathogenesis (Rue et al., 2016). Furthermore, we identified that the loss of striatal 90-kDa ribosomal S6 kinase (Rsk) is a key factor for motor, synaptic and transcription dysfunction in HD (Anglada-Huguet et al., 2016). We observed that Prostaglandin E2 EP2 activation reduces memory decline in R6/1 mice by the induction of BDNF-dependent synaptic plasticity (Anglada-Huguet et al., 2016). The interaction between BDNF and striatal-enriched protein tyrosine phosphatase (STEP) has also been characterized with the effect in normal cognition or in neuropsychiatric disorders (Saavedra et al., 2016; Xu et al., 2016). Moreover, we described the role of STEP in nociception (Azkona et al., 2016). We described the role of RTP801 as a novel downstream effector in the neurodegenerative processes activated in HD (Martin-Flores et al., 2016) and Parkinson s disease (Canal et al., 2016). In another study, we described the metabolic profiling for the identification of biomarkers in HD mouse models by on-line solid-phase extraction capillary electrophoresis mass spectrometry (Pont et al., 2016). The molecular mechanisms that we have described that could be potential therapeutic agents related to corticostriatal and hippocampal cognitive dysfunction in HD were reviewed in Puigdellivol et al., In our studies in regenerative medicine for neurodegenerative disorders, we described that forced cell cycle exit and modulation of GABAA, CREB, and GSK3β signaling promote functional maturation of induced pluripotent stem cell-derived neurons (Telezhkin et al., 2016). Furthermore, we observed that the expression of human DARPP-32 protein isoforms depends on the striatal neurodevelopmental (Straccia et al., 2016). We have also characterized the differentiation of canine gliomas towards both neurons and glial cells (Ciezka et al., 2016; Herranz et al., 2016). Keywords BDNF, neuronal plasticity, biomarkers, kinases, phosphatases, neurogenesis, regenerative medicine, ipscs Publications Lao-Peregrin C., Ballesteros J.J., Fernandez M., Zamora-Moratalla A., Saavedra A., Gomez Lazaro M. et al. Caffeine-mediated BDNF release regulates long-term synaptic plasticity through activation of IRS2 signaling. Addiction Biology Ciezka M., Acosta M., Herranz C., Canals J.M., Pumarola M., Candiota A.P. et al. Development of a transplantable glioma tumour model from genetically engineered mice: MRI/MRS/MRSI characterisation. Journal of Neuro-Oncology. 2016; Puigdellívol M, Saavedra A, Pérez-Navarro E. Cognitive Dysfunction in Huntington s Disease: Mechanisms and Therapeutic Strategies Beyond BDNF.Brain pathology (Zurich, Switzerland) Cherubini M, Ginés S. Mitochondrial fragmentation in neuronal degeneration: Toward an understanding of HD striatal susceptibility.biochemical and biophysical research communications Telezhkin V., Schnell C., Yarova P., Yung S., Cope E., Hughes A. et al. Forced cell cycle exit and modulation of GABAA, CREB, and GSK3β signaling promote functional maturation of induced pluripotent stem cell-derived neurons. American journal of physiology. Cell physiology. 2016;310(7):C520-C541. Bosch M., Fajardo A., Alcala-Vida R., Fernandez-Vidal A., Tebar F., Enrich C. et al. Hepatic primary and secondary cholesterol deposition and damage in Niemann-Pick disease. American Journal of Pathology. 2016;186(3): CIBERNED Annual Report / 109

110 Canal M., Martin-Flores N., Perez-Sisques L., Romani-Aumedes J., Altas B., Man H.-Y. et al. Loss of NEDD4 contributes to RTP801 elevation and neuron toxicity: Implications for Parkinson s disease. Oncotarget. 2016;7(37): Rue L., Banez-Corone M., Creus-Muncunill J., Giralt A., Alcala-Vida R., Mentxaka G. et al. Targeting CAG repeat RNAs reduces Huntington s disease phenotype independently of huntingtin levels. Journal of Clinical Investigation. 2016;126(11): van der Kooij M.A., Masana M., Rust M.B., Muller M.B. The stressed cytoskeleton: How actin dynamics can shape stress-related consequences on synaptic plasticity and complex behavior. Neuroscience and Biobehavioral Reviews. 2016;62: Straccia M., Carrere J., Rosser A.E., Canals J.M. Human t-darpp is induced during striatal development. Neuroscience. 2016;333: Anglada-Huguet M., Giralt A., Rue L., Alberch J., Xifro X. Loss of striatal 90-kDa ribosomal S6 kinase (Rsk) is a key factor for motor, synaptic and transcription dysfunction in Huntington s disease. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2016;1862(7): Brito V, Ginés S. p75ntr in Huntington s disease: beyond the basal ganglia.oncotarget. 2016;7(1). Research projects Code: RD12/0019/0008. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas (coordinador de red: José María Moraleda). CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RD16/0011/0017. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas. CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RTC Title: Evaluación de la eficacia de inhibidores epigenéticos en modelos experimentales de patologías humana. Principal Investigator: Merce Pallas (G402)/ Esther Pérez (G301). CIBERNED s collaboration: Yes. CIBERNED groups: G402, G301. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: ,25. Duration: 2016 Code: RTC Title: Optimización y desarrollo de un agente terapéutico basado en ácidos nucleicos para el tratamiento de la enfermedad de Huntington. Principal Investigator: Assumpcio Bosch (G607)/Esther Pérez (G301). CIBERNED s collaboration: Yes. CIBERNED groups: G607, G301. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: ,92. Duration: Code: A4530. Title: Stem Cell Differentiation JSC Joint collaborative effort between the IDIBAPS (Barcelona), the University of Milano and the University of Cardiff. Principal Investigator: Josep M. Canals. CIBERNED s collaborartion: No. CIBERNED groups: G301. Type: International. Funding agency: CHDI Foundation Inc. Funding: Duration: Code: Title: Dual therapeutic benefits of isotypeselective HDAC inhibition in Huntington s disease. Principal Investigator: Silvia Ginés. CIBERNED groups: G301. Type: International. Funding agency: EHDN. Funding: Duration: CIBERNED Annual Report / 110

111 Program 2 Group: Jordi Alberch Vié Code: COMRDI Title: ADVANCECAT: acceleradora pel desenvolupament de teràpies avançades a Catalunya (ADVANCECAT: accelerating development for advanced therapies in Catalonia). Principal Investigator: Josep M. Canals. CIBERNED groups: G301. Other CIBER s collaboration: Yes (CIBERBBN). Type: CC.AA. Funding agency: Generalitat de Catalunya. Funding: Duration: Code: TECDTP Title: Custom architecturally defined 3D stem cell derived functional human neural networks for transformative progress in neuroscience and medicine (MESO_BRAIN). Principal Investigator: Jordi Soriano. CIBERNED groups: G301. Type: European. Funding agency: Comisión Europea. Funding: Duration: Code: SAF R. Title: Desarrollo, diferenciación y maduración neuronal en la enfermedad de Huntington. Principal Investigator: Josep M. Canals. CIBERNED groups: G301. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: H2020-MSCA-ITN Title: European Training Network for Cellbased Regenerative Medicine. Principal Investigator: Jordi Alberch. CIBERNED groups: G301. Type: European. Funding agency: Comisión Europea. Funding: Duration: Code: SAF R. Title: Modulación de la plasticidad sináptica como estrategia terapéutica en la enfermedad de Huntington. Principal Investigator: Jordi Alberch/Cristina Malagelada. CIBERNED groups: G301. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: 2016 Code: TV Title: Modulation of synaptic plasticity deficits as a therapeutic strategy for Huntington s disease. Principal Investigator: Jordi Alberch. CIBERNED groups: G301. Type: Private. Funding agency: Fundaciò la Maratò de TV3. Funding: Duration: 2016 Code: A Title: Studying Human MSN Differentiation from PSC using Single-Cell RNAseq and Rodent Chimeric Models. Principal Investigator: Josep M. Canals. CIBERNED groups: G301. Type: International. Funding agency: CHDi Foundation Inc. Funding: Duration: PhD dissertations Author: Marta Cherubini. Title: Study of mitochondrial dysfunction mechanisms in Huntington s disease striatal degeneration. Date: 15 January Supervisor: Silvia Ginés Padrós CIBERNED Annual Report / 111

112 GROUP Enric I. Canela Campos 201 Principal Investigator Enric I. Canela Campos Research team 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 Estefanía Moreno Guillén Postdoctoral fellow Gemma Navarro Brugal Postdoctoral fellow David Aguinaga Andrés Edgar Angelats Canals Verónica Casadó Anguera Iñigo Etayo Labiano Patricia C. Homar Ruano Mireia Medrano Moya Irene Reyes Resina M. Mar Rodríguez Ruiz Departamento de Bioquímica y Biología Molecular Facultad de Biología, Universidad de Barcelona Diagonal 643, planta Barcelona, Catalunya (Spain) Tel.: Fax: ecanela@ub.edu Jasmina Jiménez Cano Technician 2016 CIBERNED Annual Report / 112

113 Summary Our area of interest has been the GPCRs in the CNS, specially their ability to form homo- and hetero-oligomers. We focused on discovering receptor heteromers as new targets for treatment of CNS disorders. Most relevant publications in this period were: We reviewed the ability of several GPCRs to form homodimers and heteromers. Heterotetrameric structures have been proposed for dopamine and adenosine receptors (D1R D3R, A2AR A1R and A2AR-D2R). The structural model proposed for these complexes is a heteromer constituted by two receptor homodimers. We focused on the A2AR D2R heterotetramer and reviewed the interfaces involved in this and other recently reported heteromers of GPCRs. Furthermore, we discussed several published studies showing the ex vivo expression of A2AR D2R heteromers. We summarized the functional and pharmacological characteristics of D3Rs, including the design and clinical relevance of full agonists, partial agonists and antagonists, as well as the capacity of these receptors to form active homodimers, heterodimers or higher order receptor complexes as pharmacological targets in several neurological and neurodegenerative disorders. The most promising therapeutic strategy in the future is likely to involve the application of heteromer-selective drugs. We proposed drugs acting on cannabinoid CB2R as promising drugs to combat CNS diseases (PD, HD, cerebellar ataxia, amyotrohic lateral sclerosis). Differential localization of CB2R in neural cell types and upregulation in neuroinflammation are keys to understand the therapeutic potential in inter alia diseases that imply progressive neurodegeneration. We reviewed that the protomer-protomer interactions within A1R homomers account for some unexpected pharmacological characteristics that can explain some biphasic effects reported at both low and high concentration of caffeine. We proposed for the A1R-A2AR heteromer a molecular architecture formed by a rhombusshaped heterotetramer, which is bound to two different interacting heterotrimeric G proteins (Gi and Gs). We found a preferential Gi/o coupling of the ghrelin receptor GHSR1a-GHS-R1b complex in HEK cells and, unexpectedly, a preferential Gs/ olf coupling in both striatal and hippocampal neurons in culture. Experiments in HEK cells demonstrated that D1R co-expression promotes a switch in GHS-R1a-G protein coupling from Gi/o to Gs/olf, but only upon co-expression of GHS-R1b. Furthermore, BRET assays showed that D1R interacts with GHS-R1a, but only in the presence of GHS-R1b. We found extracellular striatal levels of adenosine significantly lower in HD animal models as compared with controls and striatal equilibrative nucleoside transporter (ENT1) binding sites were significantly upregulated in HD mice. ENT1 transcript was significantly upregulated in HD disease patients at an early neuropathological severity stage, but not those with a higher severity stage. This study demonstrated that ENT1 and adenosine constitute biomarkers of the initial stages of neurodegeneration in HD disease. We also reported the discovery of D1R-histamine H3R-NMDAR heteromers in heterologous systems and in rodent brain cortex. Heteromers were detected by co-immunoprecipitation and PLA in the rat cortex where H3R agonists, via negative cross-talk, and H3R antagonists, via cross-antagonism, decreased D1R agonist ERK or Akt signaling, and counteracted D1Rmediated excitotoxic cell death. Thus, H3R in D1-H3-NMDA heteroreceptors arise as promising targets to prevent neurodegeneration in AD. We assessed whether inhibiting fatty acid amine hydrolase could provide beneficial effects on the time course of PD. We demonstrated that its inhibition has anti-cataleptic effects in PD animal models that are mediated by CB1 and CB2 receptors. We also reported the discovery and synthesis of a fluorophore-conjugated CB2R-selective compound, CM-157, which was useful for pharmacological characterization of CB2R by using a TR-FRET assay. The homogeneous binding assay described, may serve to a better characterization of agonist binding to CB2R and to identify specific properties of CB2R on living cells CIBERNED Annual Report / 113

114 Keywords AD, adenosine receptors, allosteric interactions, caffeine, cannabinoids, dopamine receptors, ghrelin receptors, G-protein coupled receptors oligomerization, HD, NMDA receptors, nucleoside transporters, PD Publications Rico A.J., Dopeso-Reyes I.G., Martinez-Pinilla E., Sucunza D., Pignataro D., Roda E. et al. Neurochemical evidence supporting dopamine D1 D2 receptor heteromers in the striatum of the long-tailed macaque: changes following dopaminergic manipulation. Brain Structure and Function. 2016; Hernandez-Garzon E., Fernandez A.M., Perez-Alvarez A., Genis L., Bascunana P., Fernandez de la Rosa R. et al. The insulin-like growth factor I receptor regulates glucose transport by astrocytes. Glia Rodríguez-Ruiz M, Moreno E, Moreno-Delgado D, Navarro G, Mallol J, Cortés A et al. Heteroreceptor Complexes Formed by Dopamine D₁, Histamine H₃, and N-Methyl-D-Aspartate Glutamate Receptors as Targets to Prevent Neuronal Death in Alzheimer s Disease.Molecular neurobiology Morales P., Gomez-Canas M., Navarro G., Hurst D.P., Carrillo-Salinas F.J., Lagartera L. et al. Chromenopyrazole, a Versatile Cannabinoid Scaffold with in Vivo Activity in a Model of Multiple Sclerosis. Journal of Medicinal Chemistry. 2016;59(14): Perreault M.L., Hasbi A., Shen M.Y.F., Fan T., Navarro G., Fletcher P.J. et al. Disruption of a dopamine receptor complex amplifies the actions of cocaine. European Neuropsychopharmacology. 2016;26(9): Guitart X., Bonaventura J., Rea W., Orru M., Cellai L., Dettori I. et al. Equilibrative nucleoside transporter ENT1 as a biomarker of Huntington disease. Neurobiology of Disease. 2016;96: Oliva H, Pacheco R, Martinez-Navio JM, Rodríguez-García M, Naranjo-Gómez M, Climent N et al. Increased expression with differential subcellular location of cytidine deaminase APOBEC3G in human CD4(+) T-cell activation and dendritic cell maturation.immunology and cell biology. 2016;94(7). Valenzuela R, Costa-Besada MA, Iglesias-Gonzalez J, Perez-Costas E, Villar-Cheda B, Garrido-Gil P et al. Mitochondrial angiotensin receptors in dopaminergic neurons. Role in cell protection and aging-related vulnerability to neurodegeneration.cell death & disease. 2016;7(10). Rodrigues RJ, Almeida T, Díaz-Hernández M, Marques JM, Franco R, Solsona C et al. Presynaptic P2X1-3 and α3-containing nicotinic receptors assemble into functionally interacting ion channels in the rat hippocampus.neuropharmacology. 2016;105. Borroto-Escuela D.O., Wydra K., Pintsuk J., Narvaez M., Corrales F., Zaniewska M. et al. Understanding the functional plasticity in neural networks of the basal ganglia in cocaine use disorder: A role for allosteric receptor-receptor interactions in A2A-D2 heteroreceptor complexes. Neural Plasticity. 2016;2016. Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J. et al. A significant role of the truncated ghrelin receptor GHS-R1b in ghrelin-induced signaling in neurons. Journal of Biological Chemistry. 2016;291(25): Franco R., Martinez-Pinilla E., Lanciego J.L., Navarro G. Basic pharmacological and structural evidence for class A G-protein-coupled receptor heteromerization. Frontiers in Pharmacology. 2016;7. Cortes A., Casado-Anguera V., Moreno E., Casado V. Caffeine, Adenosine A1 Receptors, and Brain Cortex. Molecular Aspects. Neuropathology of Drug Addictions and Substance Misuse. 2016;3: CIBERNED Annual Report / 114

115 Program 2 Group: Enric I. Canela Campos Sánchez-Soto M, Bonifazi A, Cai NS, Ellenberger MP, Newman AH, Ferré S et al. Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D2-Like Receptor Agonist.Molecular pharmacology. 2016;89(4). Casadó-Anguera V, Bonaventura J, Moreno E, Navarro G, Cortés A, Ferré S et al. Evidence for the heterotetrameric structure of the adenosine A2A-dopamine D2 receptor complex. Biochemical Society transactions. 2016;44(2). Celorrio M., Fernandez-Suarez D., Rojo-Bustamante E., Echeverry-Alzate V., Ramirez M.J., Hillard C.J. et al. Fatty acid amide hydrolase inhibition for the symptomatic relief of Parkinson s disease. Brain, Behavior, and Immunity. 2016;57: Navarro G., Cordomi A., Zelman-Femiak M., Brugarolas M., Moreno E., Aguinaga D. et al. Quaternary structure of a G-protein-coupled receptor heterotetramer in complex with Gi and Gs. BMC Biology. 2016;14(1). Navarro G., Morales P., Rodriguez-Cueto C., Fernandez-Ruiz J., Jagerovic N., Franco R. Targeting cannabinoid CB2 receptors in the central nervous system. Medicinal chemistry approaches with focus on neurodegenerative disorders. Frontiers in Neuroscience. 2016;10(SEP). Cortes A., Moreno E., Rodriguez-Ruiz M., Canela E.I., Casado V. Targeting the dopamine D3 receptor: an overview of drug design strategies. Expert Opinion on Drug Discovery. 2016;11(7): Martinez-Pinilla E., Rabal O., Reyes-Resina I., Zamarbide M., Navarro G., Sanchez-Arias J.A. et al. Two affinity sites of the cannabinoid subtype 2 receptor identified by a novel homogeneous binding assay. Journal of Pharmacology and Experimental Therapeutics. 2016;358(3): Research projects Code: Title: Cannabinoid receptor heteromeric complexes as therapeutic targets in Parkinson s disease. Principal Investigator: Rafael Franco Fernández. CIBERNED s collaboration: Yes. CIBERNED groups: G201, G203. Type: Private. Funding agency: Fundació La Marató de TV3. Funding: Duration: Code: PI2016/02. Title: Monitoring the onset and evolution of neuronal dysfunctions in propagative neural disorders using microfluidic devices and translational approaches. Principal Investigator: José Antonio del Río. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G401, G201. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: 2014-SGR Title: Neurobiología Molecular. Principal Investigator: Enric I. Canela Campos. CIBERNED s collaborarion: No. CIBERNED groups: G201. Type: CC.AA. Funding agency: Generalitat de Catalunya. Funding: Duration: Code: SAF R. Title: Oligomerización de receptores adrenérgicos y dopaminérgicos en el SNC: un nuevo enfoque en el tratamiento del transtorno por déficit de atención e hiperactividad. Principal Investigator: Enric I. Canela Campos. CIBERNED groups: G201. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 115

116 Code: Title: Targeting neuronal dopamine D1- histamine H3 receptor heteromers as a new treatment of Huntington s disease. Principal Investigator: Enric I. Canela Campos. CIBERNED groups: G201. Type: Private. Funding agency: Fundació La Marató de TV3. Funding: Duration: Code: BFU R. Title: Sigma-1 receptor-containing heteromeric complexes in cocaine addiction. Principal Investigator: Rafael Franco Fernández. CIBERNED groups: G201. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 116

117 GROUP Valentín Ceña Callejo 106 Principal Investigator Valentín Ceña Research team Inmaculada Posadas Associate professor Ana Belén García Sáez Lab technician Vanesa Guijarro Lab technician Elena Galera Lab technician Laura Romero Pharmacist Darío Manzanares Pharmacist Pablo Játiva Pharmacist Cristina de la Torre Pharmacist Unidad Asociada Neurodeath Universidad de Castilla-La Mancha, Facultad de Medicina Avenida Almansa, Albacete (Spain) Tel.: CIBERNED Annual Report / 117

118 Summary The research of the Neurodeath group has been focused on two different aspects: 1) The use of nanoparticles, mainly dendrimers (but also polymers derived from cyclodextrin) to transfect genetic material, mainly sirna, into different cell types to knock down specific proteins involved in tumoral progression and neurodegenerative diseases and in the design of a special type of dendrimers that possess anti-inflammatory activity by themselves and that will be tested on animal models of multiple sclerosis. 2) To study, using both an in silico and biological approach, the molecular determinants regulating the sirna/nanoparticle interaction that provide sirna high transfection efficiency. The use of sirna will produce, in a short period of time, new generations of drugs, based on sirna, with the potential to revolutionize therapeutics in different areas. 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. Keywords Gene therapy, dendrimers, nanoparticles, neurodegeneration, sirna, transfection, glioblastoma Publications Posadas I., Monteagudo S., Cena V. Nanoparticles for brain-specific drug and genetic material delivery, imaging and diagnosis. Nanomedicine. 2016;11(7): Bravo I., Alonso-Moreno C., Posadas I., Albaladejo J., Carrillo-Hermosilla F., Cena V. et al. Phenyl-guanidine derivatives as potential therapeutic agents for glioblastoma multiforme: Catalytic syntheses, cytotoxic effects and DNA affinity. RSC Advances. 2016;6(10): Janiszewska J, Posadas I, Játiva P, Bugaj-Zarebska M, Urbanczyk-Lipkowska Z, Ceña V. Second Generation Amphiphilic Poly-Lysine Dendrons Inhibit Glioblastoma Cell Proliferation without Toxicity for Neurons or Astrocytes.PloS one. 2016;11(11). Research projects Code: PI2016/05. Title: Dream inhibitors and Alzheimer s Disease. Principal Investigator: J. Ramón Naranjo Orovio. CIBERNED s collaboration: Yes. CIBERNED groups: G307, G106, G403. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: 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 sistema de liberación de péptidos de VIH. Principal Investigator: Mª Ángeles Muñoz Fdez. CIBERNED groups: G106. Other CIBER s collaboration: Yes (Nanomedicina). Type: National. Funding agency: CYTED. Funding: Duration: Code: BFU P. Title: Efecto de dendrímeros antiinflamatorios sobre la patogénesis de la esclerosis múltiple. Principal Investigator: Valentín Ceña. CIBERNED groups: G106. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 118

119 GROUP Antonio Cuadrado Pastor 101 Principal Investigator Antonio Cuadrado Pastor Full professor Research team Isabel Lastres Becker Assistant professor Ana Isabel Rojo Sanchís Postdoctoral fellow Ángel Juan García Yagüe Postdoctoral fellow Marta Pajares Cabetas Natalia Robledinos Antón Diego Lastra Martínez Grade student Sara Castro Grade student Caroline Stokes Grade student Departamento de Bioquímica e Instituto de Investigaciones Biomédicas Facultad de Medicina, Universidad Autónoma de Madrid Arzobispo Morcillo 4, Madrid (Spain) Tel.: Fax: antonio.cuadrado@uam.es 2016 CIBERNED Annual Report / 119

120 Summary The problem Aging is the main factor contributing towards both Parkinson s (PD) and Alzheimer s (AD) diseases. These chronic diseases are incurable and their disabling effects may continue for years or even decades. Studies on animal models of AD and PD and on human postmortem brain tissues, indicate that many pathological changes in the brain derive from a network of local stresses, like oxidative stress, tightly connected to inflammatory and proteotoxic stresses. Local stressful conditions are probably challenged by pathologically modified proteins, and, through a vicious cycle, may further trigger alteration of key molecules. Our team has been studying protective mechanisms used to maintain homeostatic respones and how these mechanims coudl be targeted pharmacologically to provide superior defense. Our approach We are currently studying the role of transcription factor NRF2 in protection against stimuli that induce neurodegeneration. NRF2 is a protein that regulates the expression of about 250 genes. These genes possess the antioxidant response element (ARE) in their promoters. The genes participate in adaptive responses to oxidative, inflammatory and proteotoxic stress and in the regulation of enzymes involved in biotransformation and glutathione metabolism. Using genetically modified rodent models as well as pharmacological approaches, we are studying the contribution of this transcription factor to the protection against oxidative damage and neuroinflammation in toxic (MPTP and 6-OHDA) and genetic (alpha-synuclein) models of Parkinson s disease and in transgenic mice possessing amyloidopathy (APPV717I) and tauopathy (TauP301L), which are characteristic of Alzheimer s disease. Objectives Generation of knowledge: Understanding the mechanisms that regulate NRF2 is fundamental to determine its physiological role and its pathological alterations as well as to design new pharmacological strategies. We are currently studying the regulation of NRF2 by signaling pathways. We have already described its regulation by the GSK-3/beta-TrCP pathway. We are now analyzing its participation in cell signaling by primary cilium and proliferative stimuli. Low-grade chronic inflammation is a key element of neurodegenerative diseases. We are studying the crosstalk between NF-kB and NRF2, key elements in the pro and antiinflammatory phenotypes of microglia. Applicability: in collaboration with several companies, we are looking for novel mechanisms of regulation of NRF2 in the brain that could serve to reinforce its activity against neurotoxic stimuli. In preclinical models of Parkinson s disease, we are focusing on repurposing of dimethyl fumarate, a compound already used in clinical practice for multiple sclerosis. Keywords Oxidative Stress, Neuroinflammation, Proteinopathy, Neuroprotection, NRF2, WNT, GSK-3, TAU Publications Navarro E, Gonzalez-Lafuente L, Pérez-Liébana I, Buendia I, López-Bernardo E, Sánchez-Ramos C et al. Heme-Oxygenase I and PCG-1α Regulate Mitochondrial Biogenesis via Microglial Activation of Alpha7 Nicotinic Acetylcholine Receptors Using PNU Antioxidants & redox signaling Pajares M., Jimenez-Moreno N., Garcia-Yague A.J., Escoll M., de Ceballos M.L., Van Leuven F. et al. Transcription factor NFE2L2/NRF2 is a regulator of macroautophagy genes. Autophagy. 2016; Lastres-Becker I., Nonis D., Eich F., Klinkenberg M., Gorospe M., Kotter P. et al. Mammalian ataxin-2 modulates translation control at the pre-initiation complex via PI3K/mTOR and is induced by starvation. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2016;1862(9): CIBERNED Annual Report / 120

121 Program 2 Group: Antonio Cuadrado Pastor Cuadrado A. NRF2 in neurodegenerative diseases. Current Opinion in Toxicology. 2016; 1: Lastres-Becker I., Garcia-Yague A.J., Scannevin R.H., Casarejos M.J., Kugler S., Rabano A. et al. Repurposing the NRF2 Activator Dimethyl Fumarate as Therapy Against Synucleinopathy in Parkinson s Disease. Antioxidants and Redox Signaling. 2016;25(2): Schmidt H.H.H.W., Stocker R., Paulsen G., Ghezzi P., Riley D., Daiber A. et al. Response to I. Batinic-Haberle et al.antioxidants and Redox Signaling. 2016;24(9): Research projects Code:PI2015/02. Title: Validación de nuevas dianas terapéuticas y nuevos biomarcadores en enfermedad de Parkinson. Principal Investigator: José Luis Labandeira García. CIBERNED s collaboration: Yes. CIBERNED groups: G208, G101, G202, G203. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: SAF R. Title: Papel de NRF2 como modulador antioxidante de la neuroinflamacion en la enfermedad de Alzheimer. Principal Investigator: Antonio Cuadrado Pastor. CIBERNED groups: G101. Other CIBER s collaborarion: No. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: SAF REDT. Title: Red de investigacion en NRF2 como nodo del patogenosoma. Principal Investigator: Antonio Cuadrado. CIBERNED groups: G101. Type: National. Funding agency: Ministerio de Economía, Industria y Competitiividad. Funding: Duration: Code: PCIN Title: Advanced theranostic approach in cancer combining photodynamic therapy and nanoparticles. Principal Investigator: Luis Felipe Ferreira (Portugal). CIBERNED groups: G101. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: P_37_732. Title: Knowledge transfer in redox biology for developing advanced molecular tools in neurodegenerative diseases. Principal Investigator: Antonio Cuadrado Pastor. CIBERNED groups: G101. Type: European. Funding agency: Comisión Europea. Funding : Duration: CIBERNED Annual Report / 121

122 GROUP Isabel Fariñas Gómez 102 Principal Investigator Isabel Fariñas Full professor Research team Francisco Pérez Sánchez Lecturer Martina Kirstein Lecturer Alexandra Bizy Researcher fellow María Pimentel Santillana Researcher fellow Raquel Montalbán Anna Lozano José Manuel Morante Redolat Lecturer Sacramento Rodríguez Ferrón Ramón y Cajal researcher Ana Pérez Villalba Researcher fellow and Lecturer Salomé Sirerol Piquer Researcher fellow and Lecturer Departamento de Biología Celular Universidad de Valencia Doctor Moliner Burjassot (Spain) Tel.: , (despacho) 3246 Fax: isabel.farinas@uv.es Germán Belenguer Sánchez Beatriz Martí Prado Ana Domingo Muelas Pau Carrillo Barberá María José Palop Benlloch Technician Fabrice Durupt Technician Elba Barberà Ferragud Technician Miquel Marí Zaragoza Technician Cristina Andrés Carbonell Technician 2016 CIBERNED Annual Report / 122

123 Summary Our laboratory develops two lines of research in neurodegeneration and cell therapy: 1) 2) The study of cellular and molecular alterations underlying the dopaminergic neurodegeneration associated with Parkinson s disease. The study of neural stem cell regulation (NSCs). We use experimental mouse models in which to identify and evaluate cellular and molecular mechanisms that may underlie dopaminergic neurodegeneration or that can be used for the understanding of adult NSCs and their manipulation for therapeutic purposes. In the context of the CIBERNED program, apart from the specific studies within each of these lines, we work on aspects that combine both, i.e. those related with the regulation of adult neurogenesis in parkinsonism and during aging. During the year 2016 we have developed methodology for the analysis of populations of quiescent and activated NSCs by flow cytometry, in order to study the response of these cells to situations of injury, including peripheral lesions. We have also strengthened our studies into the effects of the synaptic regulator involved in Parkinson s disease, alpha-synuclein, in adult neurogenesis processes. Our data indicate that NSCs respond to systemic lesions by acquiring a alert state mediated and modulated by innate immunity systems and that alpha-synuclein is a regulator of the maintenance of NSCs. On the other hand, we have explored the extracellular actions of alpha-synuclein in NSCs and how it can be internalized in neurogenic niches. Part of these studies are ongoing collaborations with other CIBERNED groups, such as those of Rosario Moratalla (Instituto Cajal), Miquel Vila (Hospital Vall d Hebrón), and José López Barneo (Juan José Toledo Aral; IBiS). Keywords Neural stem cells, adult neurogenesis, Parkinson s disease, cell therapy Publications Morante-Redolat J.M., Farinas I. Fetal neurogenesis: Breathe HIF you can. EMBO Journal Belenguer G., Domingo-Muelas A., Ferron S.R., Morante-Redolat J.M., Farinas I. Isolation, culture and analysis of adult subependymal neural stem cells. Differentiation Gonzalez-Cano L., Fuertes-Alvarez S., Robledinos-Anton N., Bizy A., Villena-Cortes A., Farinas I. et al. p73 is required for ependymal cell maturation and neurogenic SVZ cytoarchitecture. Developmental Neurobiology. 2016;76(7): Porlan E., Marti-Prado B., Consiglio A., Farinas I. Stable and efficient genetic modification of cells in the adult mouse V-SVZ for the analysis of neural stem cell autonomous and non-autonomous effects. Journal of Visualized Experiments. 2016;2016(108). Research projects Code: RD12/0019/0008. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas (coordinador de red: José María Moraleda). CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RD16/0011/0017. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas. CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: CIBERNED Annual Report / 123

124 Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz Cánoves. CIBERNED s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: Convenio Botin-UV. Title: Estudio de células madre en el ámbito de las investigaciones básicas en terapia celular. Principal Investigator: Isabel Fariñas. CIBERNED groups: G102. Type: Private. Funding agency: Fundación Botín-Banco Santander. Funding: Duration: Code: SAF Title: Regulación molecular de la quiescencia: células madre neurales adultas. Principal Investigator: Isabel Fariñas. CIBERNED groups: G102. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 124

125 GROUP Rafael Fernández-Chacón 606 Principal Investigator Rafael Fernández-Chacón Associate professor Research team José Luis Muñoz Bravo Researcher José Luis Nieto González Researcher Alejandro Arroyo Saborido Technician María del Carmen Rivero Mena Technician Ángela Lavado Roldán Researcher Marina Valenzuela Villatoro Ismael Valladares Millán Master student Pablo García-Junco- Clemente PhD fellow María Isabel Reina Graduated in Basic and Experimental Biomedicine Instituto de Biomedicina de Sevilla (IBiS) Hospital Universitario Virgen del Rocío. CSIC. Universidad de Sevilla Departamento de Fisiología Médica y Biofísica. Avenida Manuel Siurot s/n, Sevilla (Spain) Tel.: Fax: rfchacon@us.es CIBERNED Annual Report / 125

126 Summary The operation of neural circuits from birth to senility depends on synaptic function. Indeed, synaptic dysfunction is a major hallmark underlying neurodegenerarive diseases. Our laboratory investigates the molecular mechanisms underlying the maintenance of synapses along the time in the mammalian brain. Cysteine String Protein-alpha (CSPalpha) is a DNAJ-containing synaptic vesicle protein that, as a co-chaperone, prevents presynaptic degeneration. In humans, mutations in the DNAJC5 gene, that codes CSP-alpha, cause autosomal-dominant adult onset neuronal ceroid lipofuscinosis, that leads to seizures and early death in young adults. In order to investigate the function of CSP-alpha in the brain in vivo we have used genetically modified mouse stem cells bearing a DNAJC5 floxed allele (European Conditional Mouse Mutagenesis Program, Skarnes et al. Nature. 474: (2011)) to generate CSPalpha conditional knock-out mice. We have observed that, in CSP-alpha fl/fl mice, CSPalpha expression is specifically abolished by Cre-recombinase activity. Importantly, those results validate the genomic manipulation at the DNAJC5 locus to generate conditional knock-out mice. We are currently breeding CSP-alpha fl/fl mice against several mouse lines that express Cre-recombinase under different neuronal specific promoters to investigate the role of CSP-alpha in the long-term maintenance of different synaptic types. Based on our unpublished results, indicating that CSP-alpha plays a cell-autonomous role in adult neural stem cell, we have extended our studies to muscle stem cells, the satellite cells. This study is part of a CIBERNED collaborative project focused on molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Furthermore, our collaboration with Dr. Carmen Parada s and other CIBERNED s groups has fruitfully contributed to understand the molecular mechanisms underlying a novel familial limb-girdle muscular dystrophy linked to a mutation in the human POGLUT1 (protein O-glucosyltransferase 1) gene that leads to a severe loss of satellite cells (Servián-Morilla, EMBO Mol Med. 8: (2016)). Keywords Synapse, synaptic vesicle cycle, hippocampus, neurogenesis, neuronal ceroid lipofuscinosis, Crelox system, satellite cells Publications Garcia-Junco-Clemente P, Ikrar T, Tring E, Xu X, Ringach DL, Trachtenberg JT et al. An inhibitory pull-push circuit in frontal cortex.nature neuroscience. 2017;. Epub Glerup S., Bolcho U., Mlgaard S., Bggild S., Vaegter C.B., Smith A.H. et al. SorCS2 is required for BDNF-dependent plasticity in the hippocampus. Molecular Psychiatry. 2016;21(12): Ringach D.L., Mineault P.J., Tring E., Olivas N.D., Garcia-Junco-Clemente P., Trachtenberg J.T. Spatial clustering of tuning in mouse primary visual cortex. Nature Communications. 2016;7. Lavado-Roldan A., Fernandez-Chacon R. Two for the Price of One: A Neuroprotective Chaperone Kit within NAD Synthase Protein NMNAT2. PLoS Biology. 2016;14(7). Servian-Morilla E., Takeuchi H., Lee T.V., Clarimon J., Mavillard F., Area-Gomez E. et al. A POGLUT1 mutation causes a muscular dystrophy with reduced Notch signaling and satellite cell loss. EMBO Molecular Medicine. 2016;8(11): Munoz-Bravo J.L., Flores-Martinez A., Herrero-Martin G., Puri S., Taketo M.M., Rojas A. et al. Loss of pancreas upon activated Wnt signaling is concomitant with emergence of gastrointestinal identity. PLoS ONE. 2016;11(10) CIBERNED Annual Report / 126

127 Program 2 Group: Rafael Fernández-Chacón Research projects Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz Cánoves. CIBERNED s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 127

128 GROUP Javier Fernández Ruiz 303 Principal Investigator Javier Fernández Ruiz Full professor Research team José Antonio Ramos Atance Full professor Mariluz Hernández Gálvez Associate professor Eva de Lago Femia Assistant professor María Gómez Ruiz Assistant professor Onintza Sagredo Ezkioga Assistant professor Mª Concepción García García Assistant professor Carmen Rodríguez Cueto Postdoctoral researcher Valentina Satta Postdoctoral researcher Sara Valdeolivas Rojas Francisco Espejo Porras María Ceprián Costoso Laura García Toscano Yolanda García Movellán Administrative assistant Departamento de Bioquímica y Biología Molecular III Facultad de Medicina, Universidad Complutense de Madrid. Avenida Complutense s/n Pabellón IV, 4ª planta, laboratorios 9, 10, 11 y Madrid (Spain) Tel.: Fax: jjfr@med.ucm.es J.Fernandez-Ruiz@ciberned.es 2016 CIBERNED Annual Report / 128

129 Summary The work during 2016 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 (Δ9-THC, cannabidiol, cannabigerol) and semisynthetic derivatives in experimental models of this disease, in particular R6/2 mice. During this last year, we have progressed in those studies addressed to determine the molecular and cellular mechanisms underlying these effects using in vitro models of this disease (Q7 versus Q111 cells), in which were studying the Nrf-2 signaling as the key mechanism. Lastly, we have already published 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 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 Δ 9-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 PPAR-γ agonists) and cannabidiol-quinones (CB2 and PPAR-γ agonists) in experimental models of this disease. We are particularly interested in the role of CB2 receptors and, more recently, in GPR55, a novel endocannabinoidrelated receptor, in relation with the control of inflammation in this disease, by comparing different experimental models and including studies in mice deficient for these receptors. In the case of amyotrophic lateral sclerosis (ALS), 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 (FTD). In the case of Vivacell Biotechnology, these studies are being carried out within a project funded in the 2014 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, in which, during 2016, we have concentrated on finishing the first study aimed at demonstrating the existence of changes in endocannabinoid signaling in the spinal cord of dogs affected by this disease, study that has been just published and that allowed the identification of novel therapeutic targets. During 2016, we have also initiated our new project funded by the MINECO-Biomedicine National Program, concentrated in ALS and FTD, which is a joint project with a group of the CSIC Medical Chemistry Institute, and addressed to the development of new therapies based on targeting GPR55, GPR18, CB2 receptors using allosteric modulators, and CB2/PPAR-γ using hybrid compounds. Lastly, in the case of spinocerebellar ataxias, the work has been concentrated in finishing the biochemical and histopathological studies addressed to demonstrate the occurrence of an endocannabinoid dysregulation in different CNS areas during the course of the disease, using SCA-3 transgenic mice, and we are carrying out pharmacological studies addressed to correct such dysregulation and to elicit neuroprotective effects, including GPR55 as a new therapeutic target. Keywords Cannabinoids, CB1 receptors, CB2 receptors, GPR55, PPAR-γ, neuroprotection, Huntington s disease, Parkinson s disease, amyotrophic lateral sclerosis/frontotemporal dementia, spinocerebellar ataxias 2016 CIBERNED Annual Report / 129

130 Publications Lopez-Sendon Moreno J.L., Garcia Caldentey J., Trigo Cubillo P., Ruiz Romero C., Garcia Ribas G., Alonso Arias M.A.A. et al. A double-blind, randomized, cross-over, placebo-controlled, pilot trial with Sativex in Huntington s disease. Journal of Neurology. 2016; Morales P, Whyte L, Chicharro R, Gómez-Cañas M, Pazos R, Goya P et al. Identification of Novel GPR55 Modulators Using Cell-Impedance-Based Label-Free Technology.Journal of medicinal chemistry Rodríguez-Cueto C, Hernández-Gálvez M, Hillard CJ, Maciel P, García-García L, Valdeolivas S et al. Dysregulation of the endocannabinoid signaling system in the cerebellum and brainstem in a transgenic mouse model of spinocerebellar ataxia type-3.neuroscience Morales P., Gomez-Canas M., Navarro G., Hurst D.P., Carrillo-Salinas F.J., Lagartera L. et al. Chromenopyrazole, a Versatile Cannabinoid Scaffold with in Vivo Activity in a Model of Multiple Sclerosis. Journal of Medicinal Chemistry. 2016;59(14): Hernandez-Folgado L, Stevenson LA, Morales P, Gómez-Cañas M, Pazos MR, Cascio MG. et al. Exploring the Benzimidazole Ring as a Substitution for Indole in Cannabinoid Allosteric Modulators. Cannabis and Cannabinoid Research. 2016; 1(1): Navarro G., Morales P., Rodriguez-Cueto C., Fernandez-Ruiz J., Jagerovic N., Franco R. Targeting cannabinoid CB2 receptors in the central nervous system. Medicinal chemistry approaches with focus on neurodegenerative disorders. Frontiers in Neuroscience. 2016;10(SEP). Del Río C, Navarrete C, Collado JA, Bellido ML, Gómez-Cañas M, Pazos MR et al. The cannabinoid quinol VCE alleviates bleomycin-induced scleroderma and exerts potent antifibrotic effects through peroxisome proliferator-activated receptor-γ and CB2 pathways. Scientific reports. 2016;6. Deiana V, Gómez-Cañas M, Pazos MR, Fernández-Ruiz J, Asproni B, Cichero E et al. Tricyclic pyrazoles. Part 8. Synthesis, biological evaluation and modelling of tricyclic pyrazole carboxamides as potential CB₂ receptor ligands with antagonist/inverse agonist properties.european journal of medicinal chemistry. 2016;112. Diaz-Alonso J., Paraiso-Luna J., Navarrete C., Del Rio C., Cantarero I., Palomares B. et al. VCE-003.2, a novel cannabigerol derivative, enhances neuronal progenitor cell survival and alleviates symptomatology in murine models of Huntington s disease. Scientific Reports. 2016;6. Rubio M, Valdeolivas S, Piscitelli F, Verde R, Satta V, Barroso E et al. Analysis of endocannabinoid signaling elements and related proteins in lymphocytes of patients with Dravet syndrome. Pharmacology research & perspectives. 2016;4(2). Gomez-Canas M., Morales P., Garcia-Toscano L., Navarrete C., Munoz E., Jagerovic N. et al. Biological characterization of PM226, a chromenoisoxazole, as a selective CB2 receptor agonist with neuroprotective profile. Pharmacological Research. 2016;110: Gómez-Gálvez Y, Palomo-Garo C, Fernández-Ruiz J, García C. Potential of the cannabinoid CB(2) receptor as a pharmacological target against inflammation in Parkinson s disease. Progress in neuro-psychopharmacology & biological psychiatry. 2016;64. Palomo-Garo C., Gomez-Galvez Y., Garcia C., Fernandez-Ruiz J. Targeting the cannabinoid CB2 receptor to attenuate the progression of motor deficits in LRRK2-transgenic mice. Pharmacological Research. 2016;110: Research projects Code: PI2016/04. Title: The ALS CIBERNED challenge: Accelerating new drug discovery. Principal Investigator: Adolfo López De Munain Arregui. CIBERNED s collaboration: Yes. CIBERNED groups: G609, G303, G503, G408. Type: Intramurales. Funding agency: CIBERNED Funding: Duration: CIBERNED Annual Report / 130

131 Program 2 Group: Javier Fernández Ruiz Code: RTC Title: Desarrollo preclínico de nuevos cannabinoides para el tratamiento de esclerodermia y esclerosis lateral amiotrófica. Principal Investigator: Javier Fernández Ruiz. CIBERNED groups: G303. Type: National. Funding agency: Ministerio Economía, Industria y Competitividad. Funding: Duration: Code: FIS PIE13/ Title: Modulation of cellular micro RNAs as a therapeutic strategy for he cure of HIV infection. Principal Investigator: Santiago Moreno. CIBERNED groups: G303. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: GW2015. Title: Preclinical development of phytocannabinoid-based therapies for the treatment of disease progression in amyotrophic lateral sclerosis/frontotemporal dementia using TDP-43 transgenic mice. Principal Investigator: Javier Fernández Ruiz, Eva de Lago Femia. CIBERNED groups: G303. Type: International. Funding agency: GW Research Ltd. (UK). Funding: Duration: Code: SAF C2-1-R. Title: Dianas del sistema endocannabinoide para el desarrollo de terapias frente a la neurodegeneración: énfasis en la ELA y otras enfermedades neurodegenerativas. Principal Investigator: Javier Fernández Ruiz y Eva de Lago Femia. CIBERNED groups: G303. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: 2016 PhD dissertations Author: María Gómez Cañas. Title: Caracterización de nuevos ligandos cannabinoides con potencial neuroprotector. Date: 3 June Supervisor: Javier Fernández Ruiz. Author: Cristina Palomo Garo. Title: Análisis del sistema endocannabinoide en la enfermedad de Parkinson: hacia un tratamiento polivalente. Date: 8 November Supervisor: Javier Fernández Ruiz CIBERNED Annual Report / 131

132 GROUP José Manuel Fuentes Rodríguez 103 Principal Investigator José Manuel Fuentes Rodríguez Full professor Research team Rosa Ana González Polo Researcher Mireia Niso Santano Researcher Sokhna Maryama Seydina Yakhine DIOP PhD, CIBERNED Mario Rodríguez Arribas, FPU Ignacio Casado Naranjo Head of department Elisabet Uribe Carretero Guadalupe Martínez Chacón Technician CIBERNED María Pura Luceño Technician Departamento de Bioquímica y Biología Molecular y Genética Facultad de Enfermería y Terapia Ocupacional Universidad de Extremadura. Avenida de la Universidad s/n Cáceres (Spain) Tel.: (ext 51286) Fax: jfuentes@unex.es 2016 CIBERNED Annual Report / 132

133 Summary Our lines of work during 2016 have continued to focus on the study of basic molecular mechanisms related to the etiopathogenesis of Parkinson s disease, regulation of autophagy and role of proteins derived from PARK genes in both processes. On the other hand we have also continued our collaboration and provision of services to the Extremadura Health Service in the molecular diagnosis of Parkinson s disease. Following the work carried out since 2009 in collaboration with the group of Dr. Adolfo López de Munain (CIBERNED), we have continued with characterization of the basal deregulation of macroautophagy in fibroblasts from Parkinson s disease patients with the pathogenic G2019S mutation in LRRK2. Fibroblasts from individuals with the G2019S mutation of LRRK2 have autophagy levels that are substantially higher than those observed in fibroblasts from individuals without such mutation. This exacerbation is accompanied by a greater sensitivity of these cells to toxins, such as MPP +, related to Parkinson s disease. We are also advancing functional studies using fibroblasts from individuals with the LRRK2 mutation R1441G. In this case we have also observed one of accelerated basal autophagy, however they do not present greater degradation than the fibroblasts coming from individuals without this mutation. In both models we started the analysis of the expression of enzymes type acetyltrasnferase and deacetylase and the degree of acetylation of proteins related to autophagy, since this posttraductional labeling is becoming evident as a key regulator in the macroautophagic response. In this sense, a complete screening of both types of enzymes was performed, observing significant differences between the control groups, idiopathic patients and patients with both mutations, G2019S and R1441G, especially in the latter group. Our group has also begun screening for possible neuroprotectors present in natural compounds and mediated by a modification in cellular autophagic activity. As an intraciberned collaboration we are developing a Cooperative Project (PI2015 / 03) together with the Groups of Dr. Pérez Tur and Dr. Pérez Castillo (with the participation of Dr. López de Munain) in which we are addressing the study of differential metabolic profiles in Parkinson s disease. During 2016 we have been obtained biological samples of human origin and established the colonies of genetically modified mice that we will use (which must be aged for 12 months to show parkinsonian phenotype) Finally, we have continued to be interested in the study of general mechanisms of autophagy regulation. This part of our activity is still being carried out in collaboration with the laboratory of Dr. Guido Kroemer of Apoptosis, Cancer & Immunity Laboratory, INSERM Cordeliers Research Center, Paris, France. In relation to our collaboration with the Extremadura Health Service emphasize that we have signed a service agreement in relation to the molecular diagnosis of Parkinson s disease. 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, metabolism Publications Labrador Gomez P.J., Gonzalez Sanchidrian S., Fuentes Rodriguez J.M., Gomez-Martino Arroyo J.R. Trend of lipid profile in general population from Caceres Health Area. Hipertension y Riesgo Vascular. 2017;34(1): Epub Rodríguez-Arribas M, Yakhine-Diop SM, Pedro JM, Gómez-Suaga P, Gómez-Sánchez R, Martínez-Chacón G et al. Mitochondria-Associated Membranes (MAMs): Overview and Its Role in Parkinson s Disease.Molecular neurobiology González-Polo RA, Pizarro-Estrella E, Yakhine-Diop SM, Rodríguez-Arribas M, Gómez-Sánchez R, Casado-Naranjo I. et al. The Basics of Autophagy. In Autophagy Networks in Inflammation. Springer International Publishing, 2016, pp ISBN: CIBERNED Annual Report / 133

134 Romero Sevilla R., Portilla Cuenca J.C., Lopez Espuela F., Redondo Penas I., Bragado Trigo I., Yerga Lorenzana B. et al. A stroke care management system prevents outcome differences related to time of stroke unit admission. Neurologia. 2016;31(3): Casares-Vivas M., Portilla-Cuenca J.C., Gallego-Texeira I., Calderon-Pecellin A., Gallego-Curto E., Casado-Naranjo I. Anti-NMDA antibody encephalitis secondary to herpes simplex virus infection. Medicina Intensiva. 2016;40(3): Lopez-Espuela F., Pedrera-Zamorano J.D., Jimenez-Caballero P.E., Ramirez-Moreno J.M., Portilla-Cuenca J.C., Lavado-Garcia J.M. et al. Functional status and disability in patients after acute stroke: A longitudinal study. American Journal of Critical Care. 2016;25(2): Gomez-Sanchez R., Bravo-San Pedro J.M., Gegg M.E., Gonzalez-Polo R.A., Fuentes J.M. Mitochondria: Key Organelle in Parkinson s Disease. Parkinson s Disease. 2016;2016. Gomez-Sanchez R., Yakhine-Diop S.M.S., Rodriguez-Arribas M., Bravo-San Pedro J.M., Martinez- Chacon G., Uribe-Carretero E. et al. mrna and protein dataset of autophagy markers (LC3 and p62) in several cell lines. Data in Brief. 2016;7: Rodríguez-Arribas M, Pedro JM, Gómez-Sánchez R, Yakhine-Diop SM, Martínez-Chacón G, Uribe-Carretero E. et al. Pompe Disease and Autophagy: Partners in Crime, or Cause and Consequence?. Current medicinal chemistry. 2016;23(21). Research projects Code: 2015/03. Title: Perfiles metabólicos diferenciales en enfermedad de Parkinson. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED s collaboration: Yes. CIBERNED groups: G103, G110, G209. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: PI15/ Title: Alteraciones del perfil metabolico inducidas por mutaciones patogénicas de LRRK2 como biomarcadores de la enfermedad de Parkinson. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED groups: G103. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: PI14/ Title: Búsqueda de nuevas dianas terapéuticas en la enfermedad de Parkinson. Análisis de los sistemas de reciclaje celular y estado mitocondrial en células procedentes de pacientes. Principal Investigator: Rosa Ana González Polo. CIBERNED groups: G103. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: GR Title: Ayudas para el fortalecimiento de los grupos de investigación de Extremadura. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED groups: G103. Type: CC.AA. Funding agency: Gobierno de Extremadura. Funding: ,36. Duration: CIBERNED Annual Report / 134

135 Program 2 Group: José Manuel Fuentes Rodríguez Code: BFU REDT. Title: Red de Excelencia para el estudio de la Autofagia. Principal Investigator: Patricia Boya. CIBERNED groups: G103. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: PEJ-2014-A Title: Ayudas para la Promoción de Empleo Joven e Implantación de la Garantía Juvenil en I+D+i. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED groups: G103. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: PhD dissertation Author: Raúl Mauricio Romero Sevilla. Title: Factores de naturaleza vascular y marcadores de inflamación como predictores de progresión a demencia en pacientes con deterioro cognitivo leve. Date: 6 June Supervisor: José Manuel Fuentes Rodríguez. Author: Aurora Alvarado Pacheco. Title: Estimación de la prevalencia de la enfermedad de Parkinson en Extremadura. Date: 6 April Supervisor: José Manuel Fuentes Rodríguez. Author: Sokhna MS Yakhine DIOP. Title: Acetilación, autofagia y enfermedad de Parkinson. Date: 11 March Supervisor: José Manuel Fuentes Rodríguez CIBERNED Annual Report / 135

136 GROUP José Manuel García Verdugo 113 Principal Investigator José Manuel García Verdugo Full professor Research team Vicente Herranz Pérez Postdoctoral fellow Sara García Gil-Perotín Postdoctoral fellow Susana González Granero Researcher María Durán Moreno Arantxa Cebrián Silla Technician Laboratorio de Neurobiología Comparada (SS7) Instituto Cavanilles de Biodiversidad y Biología Evolutiva. Universidad de Valencia c/catedrático José Beltrán, nº Paterna, Valencia (Spain) Tel.: Fax: j.manuel.garcia@uv.es 2016 CIBERNED Annual Report / 136

137 Summary The main line of research of our group is focused on the study of adult neurogenesis in several species of mammals, with special emphasis on the regenerative potential of this process in pathologies of the nervous system. There are notable differences between vertebrate species in both, the abundance and temporality, of adult neurogenesis. This fact led us to perform, over the last few years, a comparative study among higher vertebrates, in which a progressive decline in the presence and proliferative activity of neural stem cells is reflected in primates, and especially in the human brain. Because of these particularities, we are currently directing most of our work to the study of this process in the human brain. After first describing a migratory stream to the olfactory bulb, we have now described a migration to the ventral prefrontal cortex, not described to date. This interesting discovery has revealed that, in the human species, there is an extensive neural migration to the frontal lobe of newborns, which takes place for several months after birth. In these first months of life is when the child begins to interact with their environment and is a critical period for correct brain development. The frontal lobe of the human brain is involved in social behavior and function execution. Young neurons migrate tangentially by forming chains near the walls of the lateral ventricles, and along the outer walls of blood vessels. After migrating great distances, they differentiate and integrate in the frontal cortex of the brain, expressing inhibitory neuronal markers. The postnatal addition of these large populations of inhibitory neurons may contribute to the maturation and plasticity of the prefrontal cortex, whereas defects in the migration of these neurons could be the cause of neurodevelopmental pathologies. In addition to the neurogenesis associated with the area adjacent to the lateral ventricles of the brain, we are also interested in the study of other ventricular compartments where it is speculated that neural stem cells could reside. In the third ventricle, we performed a study on tanicite populations, of which we have described new subtypes based on the basal bodies of cilia and the expression of vimentin. Our interest in pathologies of the human brain, also led us to study of animal models of human disease, for example, Alzheimer s. In this regard, we have studied the effect of a molecule named genistein, which seems to have a beneficial effect on cognition, brain histology, and PET. This work allowed to start a clinical trial to study the effects of this molecule in Alzheimer s patients. On the other hand, we have published a study on the potential therapeutic effect derived from intraventricular injection of mesenchymal stem cells in murine models of demyelination. We have also contributed to the optimization of spinal cord MRI protocols for the assessment of amyotrophic lateral sclerosis models. In summary, we are trying to transfer the knowledge acquired throughout the research group s track record, to cell therapy and regenerative medicine applications that allow to improve the diagnosis, treatment, and prevention of neurodegenerative diseases. Keywords Neurogenesis, neural stem cells, subventricular zone, oligodendrogenesis, Alzheimer s disease, amyotrophic lateral sclerosis Publications Garcia-Gonzalez D., Murcia-Belmonte V., Esteban P.F., Ortega F., Diaz D., Sanchez-Vera I. et al. Erratum to: Anosmin-1 over-expression increases adult neurogenesis in the subventricular zone and neuroblast migration to the olfactory bulb. Brain Structure and Function. 2016; 1-. Gil-Perotin S, Cebrian-Silla A, Herranz-Pérez V, Garcia-Belda P, Gil-García S, Fil M, Lee JS, Nachury MV, Garcia-Verdugo JM. Localization of GFP-tagged proteins at the electron microscope. In: Receptor and ion channel detection in the brain. Luján R, Ciruela F, editors. Springer Science+Business Media New York, 2016, Vol. 110, pp ISBN: Molina-Navarro MM, García-Verdugo JM. Neurobiology. In: Adult neurogenesis in the hippocampus, Health, Psychopathology, and Brain Disease. Canales JJ, editors. Elsevier, 2016, pp ISBN: CIBERNED Annual Report / 137

138 Paredes M.F., Sorrells S.F., Garcia-Verdugo J.M., Alvarez-Buylla A. Brain size and limits to adult neurogenesis. Journal of Comparative Neurology. 2016;524(3): Ogino T., Sawada M., Takase H., Nakai C., Herranz-Perez V., Cebrian-Silla A. et al. Characterization of multiciliated ependymal cells that emerge in the neurogenic niche of the aged zebrafish brain. Journal of Comparative Neurology. 2016;524(15): Bonet-Costa V., Herranz-Perez V., Blanco-Gandia M., Mas-Bargues C., Ingles M., Garcia-Tarraga P. et al. Clearing amyloid-β through PPARγ/ApoE Activation by Genistein is a Treatment of Experimental Alzheimer s Disease. Journal of Alzheimer s Disease. 2016;51(3): Figini M., Scotti A., Marcuzzo S., Bonanno S., Padelli F., Moreno-Manzano V. et al. Comparison of diffusion MRI acquisition protocols for the in vivo characterization of the mouse spinal cord: Variability analysis and application to an amyotrophic lateral sclerosis model. PLoS ONE. 2016;11(8). Paredes M.F., James D., Gil-Perotin S., Kim H., Cotter J.A., Ng C. et al. Extensive migration of young neurons into the infant human frontal lobe. Science. 2016;354(6308). Naldaiz-Gastesi N., Goicoechea M., Alonso-Martin S., Aiastui A., Lopez-Mayorga M., Garcia- Belda P. et al. Identification and Characterization of the Dermal Panniculus Carnosus Muscle Stem Cells. Stem Cell Reports. 2016;7(3): Torres-Vega E., Duran-Moreno M., Sanchez del Pino M., Yanez Y., Canete A., Castel V. et al. Immunoproteomic studies on paediatric opsoclonus-myoclonus associated with neuroblastoma. Journal of Neuroimmunology. 2016;297: Capilla-Gonzalez V., Bonsu J.M., Redmond K.J., Garcia-Verdugo J.M., Quinones-Hinojosa A. Implications of irradiating the subventricular zone stem cell niche. Stem Cell Research. 2016;16(2): Cruz-Martinez P, González-Granero S, Molina-Navarro MM, Pacheco-Torres J, García-Verdugo JM, Geijo-Barrientos E et al. Intraventricular injections of mesenchymal stem cells activate endogenous functional remyelination in a chronic demyelinating murine model.cell death & disease. 2016;7. Dadsetan S., Balzano T., Forteza J., Cabrera-Pastor A., Taoro-Gonzalez L., Hernandez-Rabaza V. et al. Reducing peripheral inflammation with infliximab reduces neuroinflammation and improves cognition in rats with hepatic encephalopathy. Frontiers in Molecular Neuroscience. 2016;9(NOV2016). Rosillo J.C., Torres M., Olivera-Bravo S., Casanova G., Garcia-Verdugo J.M., Fernandez A.S. Telencephalic-olfactory bulb ventricle wall organization in Austrolebias charrua: Cytoarchitecture, proliferation dynamics, neurogenesis and migration. Neuroscience. 2016;336: Proyectos de investigación Code: RD12/0019/0008. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas (coordinador de red: José María Moraleda). CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RD16/0011/0017. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas. CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: 2015/01-1. Title: Terapia génica dirigida a neuregulinas para el tratamiento de la degeneración de las motoneuronas en la esclerosis lateral amiotrófica. Principal Investigator: Xavier Navarro. CIBERNED s collaboration: Yes. CIBERNED groups: G607, G113, G406, G CIBERNED Annual Report / 138

139 Program 2 Group: José Manuel García Verdugo Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: RD12/0019/0016. Title: Red de terapia celular: NEUROCEL-ELA. Nodo Valencia. Principal Investigator: José Manuel García Verdugo. CIBERNED s collaboration: Yes. CIBERNED groups: G113, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: PROMETEOII/2014/075. Title: Celulas madre adultas: Aplicaciones en terapia regenerativa. Principal Investigator: José Manuel García Verdugo. CIBERNED groups: G113. Type: CC.AA. Funding agency: Generalitat Valenciana. 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. CIBERNED groups: G113. Type: Private. Funding agency: Fundació la Marató de TV3. Funding: Duration: Code: BFU P. Title: Estudio del cerebro humano: caracterización de nichos neurogénicos y nuevas vías de migración. Principal Investigator: José Manuel García Verdugo. CIBERNED groups: G113. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: PhD dissertations Author: María Durán Moreno. Title: Estudio de la organización celular y capacidad proliferativa en el tercer ventrículo de mamíferos adultos. Date: 21 January Supervisor: José Manuel García Verdugo. Author: Lucía Galán Dávila. Title: Estudio de la neurogénesis en necropsias procedentes de pacientes con esclerosis lateral amiotrófica. Date: 14 January Supervisor: José Manuel García Verdugo. Author: Romana García Gil. Title: Papel neuroprotector de las células mesenquimales humanas a nivel ocular en la esclerosis múltiple. Date: 14 January Supervisor: José Manuel García Verdugo. Author: Carmen Bellver Estellés. Title: Aislamiento y caracterización de distintas poblaciones celulares en el tejido adiposo humano. Date: 9 February Supervisor: José Manuel García Verdugo CIBERNED Annual Report / 139

140 GROUP José González Castaño 104 Principal Investigator José González Castaño Research team Raúl Sánchez Lanzas Predoctoral researcher Silvia Carlés González Researcher technician Departamento de Bioquímica e Instituto de Investigaciones Biomédicas Alberto Sols Univerdad Autónoma de Madid-CSIC. Laboratorio B13. Facultad de Medicina UAM Arzobispo Morcillo s/n Madrid (Spain) Tel.: Fax: joseg.castano@uam.es 2016 CIBERNED Annual Report / 140

141 Summary Proteasomes and lysosomes play a critical role in protein degradation and proteostasis in the cell. Three major pathways have been shown to be relevant in protein degradation in Parkinson (PD) and other neurodegenerative diseases; ubiqutin-proteasome, chaperon mediated autophagy (CMA) and macroautophagy Alpha-synuclein, a protein accumulated and aggregated in PD, has been reported to be degraded by the CMA pathway through its direct binding to the lysosomal receptor Lamp-2A. We have reported last year that in a lymphoblastoid cell line derived from a patient with Danon disease, lacking the expression of the different Lamp-2 isoforms including Lamp- 2A, the degradation of several selective substrates of the CMA pathway including alpha-synuclein, is not impaired and there is no significant changes in the steady-state protein and mrna levels compared to controls. Our work and previous work from other authors, clearly questioned the role of CMA in lysosomal protein degradation. Recently it has been reported that DJ-1, a gene whose mutation is known to be responsible of familiar autosomal recessive forms of PD, is also degraded by the CMA pathway. In our ongoing studies of the pathways of degradation of DJ-1 wild type and point mutants we are also analyzing the possible involvement of CMA pathway in DJ-1 degradation. Our work is contributing to understand and clarify the role of the different pathaways of protein degradation in neurodegenerative disease, specially in connection with PD pathology. Keywords Synuclein, DJ-1, proteasome, macroautophagy, chaperon-mediated-autophagy, proteolysis, proteostasis, Parkinson Research projects Code: SAF_2012_ Title: Papel de la proteostasis de alfasinucleina y DJ-1 en la función celular y su relación con la patogénesis de la enfermedad de Parkinson. Principal Investigator: José González Castaño. CIBERNED groups: G104. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: S2010_BMD. Title: Redes de senalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas. Principal Investigator: José González Castaño. CIBERNED s collaboration: Yes. CIBERNED groups: G111, G401, G104, G307, G409. Other CIBER s collaboration: Yes (CIBERER). Type: CC.AA. Funding agency: Comunidad de Madrid. Funding: Duration: Code: BM1307. Title: 2013 COST action proteostasis. Principal Investigator: Manuel S. Rodríguez, Rosa Barrios (Spain). CIBERNED groups: G140. Type: European. Funding agency: UE. Funding: Duration: CIBERNED Annual Report / 141

142 GROUP Manuel Guzmán Pastor 305 Principal Investigator Manuel Guzmán Pastor Full professor Research team Ismael Galve Roperh Associate professor Guillermo Velasco Díez Associate professor Cristina Blázquez Ortiz Associate professor Luigi Bellocchio Postdoctoral fellow Javier Díaz Alonso Postdoctoral fellow Raquel Bajo Grañeras Postdoctoral fellow Anna Paola Chiarlone Postdoctoral fellow Adán de Salas Quiroga Andrea Ruiz Calvo Daniel García Rincón Juan Paraíso Luna Irene Berenice Maroto Martínez José Aguareles Gorines Carlos Costas Insua Eva Resel Mariné Lab manager Departamento de Bioquímica y Biología Molecular I Facultad de Ciencias Químicas Universidad Complutense Madrid (Spain) Tel.: Fax: mguzman@quim.ucm.es Elena García Taboada Lab technician 2016 CIBERNED Annual Report / 142

143 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 2016 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) 3) 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. 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 2016, 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 Lopez-Sendon Moreno J.L., Garcia Caldentey J., Trigo Cubillo P., Ruiz Romero C., Garcia Ribas G., Alonso Arias M.A.A. et al. A double-blind, randomized, cross-over, placebo-controlled, pilot trial with Sativex in Huntington s disease. Journal of Neurology. 2016; Hernandez-Tiedra S., Fabrias G., Davila D., Salanueva I.J., Casas J., Montes L.R. et al. Dihydroceramide accumulation mediates cytotoxic autophagy of cancer cells via autolysosome destabilization. Autophagy. 2016; Andradas C., Blasco-Benito S., Castillo-Lluva S., Dillenburg-Pilla P., Diez-Alarcia R., Juanes- Garcia A. et al. Activation of the orphan receptor GPR55 by lysophosphatidylinositol promotes metastasis in triple-negative breast cancer. Oncotarget. 2016;7(30): Perucho J., Gomez A., Munoz M.P., de Yebenes J.G., Mena M.A., Casarejos M.J. Trehalose rescues glial cell dysfunction in striatal cultures from HD R6/1 mice at early postnatal development. Molecular and Cellular Neuroscience. 2016;74: Velasco G., Sanchez C., Guzman M. Anticancer mechanisms of cannabinoids. Current Oncology. 2016;23:S23-S CIBERNED Annual Report / 143

144 Chiarlone A., Borner C., Martin-Gomez L., Jimenez-Gonzalez A., Garcia-Concejo A., Garcia- Bermejo M.L. et al. MicroRNA let-7d is a target of cannabinoid CB1 receptor and controls cannabinoid signaling. Neuropharmacology. 2016;108: Bellocchio L., Ruiz-Calvo A., Chiarlone A., Cabanas M., Resel E., Cazalets J.-R. et al. Sustained Gq-protein signaling disrupts striatal circuits via JNK. Journal of Neuroscience. 2016;36(41): Diaz-Alonso J., Paraiso-Luna J., Navarrete C., Del Rio C., Cantarero I., Palomares B. et al. VCE-003.2, a novel cannabigerol derivative, enhances neuronal progenitor cell survival and alleviates symptomatology in murine models of Huntington s disease. Scientific Reports. 2016;6. Research projects Code: SAF R. Title: Identificación y caracterización de subpoblaciones del receptor CB1 cannabinoide con actividad neuroprotectora. Principal Investigator: Manuel Guzmán Pastor. CIBERNED groups: G305. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: RTC Title: Desarrollo preclínico de VCE para el tratamiento de la enfermedad de Huntington (CANNADERIV). Principal Investigator: Ismael Galve-Roperh. CIBERNED grouos: G305. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: PI15/ Title: Papel de la autofagia en cáncer: mecanismos de muerte mediada por autofagia en células tumorales y participación de genes reguladores de la autofagia en el control de la tumorigénesis. Principal Investigator: Guillermo Velasco Diez. CIBERNED groups: G305. Type: National. Funing agency: Instituto de Salud Carlos III. Funding: Duration: Code: PI15/ Title: Papel del sistema endocannabinoide en malformaciones del desarrollo cortical asociadas a epilepsia refractaria. Principal Investigation: Ismael Galve Roperh. CIBERNED groups: G305. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: CIBERNED Annual Report / 144

145 GROUP Teresa Iglesias Vacas 111 Principal Investigator Teresa Iglesias Vacas CSIC Research team Lucía García Guerra Postdoctoral researcher Ana Mª del Puerto del Pino Postdoctoral researcher Andrea Gamir Morralla Postdoctoral researcher Álvaro Sebastián Serrano Postdoctoral researcher Julia Pose Utrilla Ángel García Aldea Master student Alicia Belmonte Alfaro Grade student Sara Valle Santos FP2 student Instituto de Investigaciones Biomédicas Alberto Sols CSIC-UAM y CIBERNED Departamento de Fisiología Endocrina y del Sistema Nervioso c/arturo Duperier, nº4, Madrid (Spain) Tel.: Fax: tiglesias@iib.uam.es 2016 CIBERNED Annual Report / 145

146 Summary During last year, our group has continued investigating the role played by protein kinase D (PKD) and its substrate Kidins220 in different neuropathologies. After discovering that Kidins220 is crucial for neuronal survival we have continued analysing the importance of PKD, its phosphorylating kinase, in excitotoxic neuronal death. This type of neuronal death is at the bases of neurodegenerative processes taking place in neurological diseases with acute neuronal damage, such as cerebral ischemia, or chronic neurodegeneration, such as Parkinson s, Huntington s or Alzheimer s diseases. We have identified changes in PKD activity in in vitro and in vivo models of excitotoxicity and in several acute and chronic neurodegenerative diseases. We have characterized some of the molecular mechanisms contributing to the observed changes in the kinase activity, and how these changes directly contribute to cellular processes that lead to neuronal death. In addition, we have designed a neuroprotective strategy using lentiviral particles to transduce specifically neurons with a PKD mutant, that confers neuroprotection in in vitro and in vivo models of excitotoxicity. Part of these studies have been performed within the cooperative project CIBERNED 2015/02-06, started during this year. We will continue our research to determine the contribution of the observed modifications in Kidins220 and PKD to the etiopathology of neurodegenerative diseases, and to design novel neuroprotective strategies. Previously, we found that Kidins220 is increased in brain necropsies of patients with Alzheimer s disease (AD), where it accumulates with hyperphosphorylated tau, one of the two main neuropathological markers of this disease. This year we have published a work, developed in the context of cooperative project CIBERNED 2013/07, where we detect a positive correlation between Kidins220 and tau in brain and cerebrospinal fluid from AD patients (Gamir- Morralla et al, J Alzheimers Dis, 2017). This study highlights the potential of Kidins220 as a novel CSF biomarker in AD, something that could have a important clinical and diagnostic value. Keywords Kidins220, protein kinase D (PKD), neuroprotection, neurotoxicity, neuronal survival, parkinson, huntington, alzheimer Publications Fernandez A.M., Hernandez-Garzon E., Perez-Domper P., Perez-Alvarez A., Mederos S., Matsui T. et al. Insulin regulates astrocytic glucose handling through cooperation with IGF-I. Diabetes. 2017;66(1): Epub Gamir-Morralla A, Belbin O, Fortea J, Alcolea D, Ferrer I, Lleó A et al. Kidins220 Correlates with Tau in Alzheimer s Disease Brain and Cerebrospinal Fluid.Journal of Alzheimer s disease : JAD. 2017;55(4). Epub Gamir-Morralla A, López-Menéndez C, Medina M, Iglesias T. A novel neuroprotection target with distinct regulation in stroke and Alzheimer s disease. In: Neuroprotection in Alzheimer s Disease. Gozes I, editors. Elsevier, ISBN: Rodriguez-Tornos F.M., Briz C.G., Weiss L.A., Sebastian-Serrano A., Ares S., Navarrete M. et al. Cux1 Enables Interhemispheric Connections of Layer II/III Neurons by Regulating Kv1-Dependent Firing. Neuron. 2016;89(3): Sebastián-Serrano Á, Engel T, de Diego-García L, Olivos-Oré LA, Arribas-Blázquez M, Martínez- Frailes C et al. Neurodevelopmental alterations and seizures developed by mouse model of infantile hypophosphatasia are associated with purinergic signalling deregulation.human molecular genetics. 2016;25(19) CIBERNED Annual Report / 146

147 Program 2 Group: Teresa Iglesias Vacas Research projects Code: S2010_BMD. Title: Redes de senalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas. Principal Investigator: José González Castaño. CIBERNED s collaboration: Yes. CIBERNED groups: G111, G401, G104, G307, G409. Other CIBER s collaboration: Yes (CIBERER). Type: CC.AA. Funding agency: Comunidad de Madrid. Funding: Duration: Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz Cánoves. CIBERNED s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: SAF P. Title: Participación y regulación de PKD y KIDINS220 en los procesos y rutas moleculares de la neurodegeneración. Principal Investigator: Teresa Iglesias Vacas. CIBERNED groups: G111. Type: National. Funding agency: Ministerio de Economía y Competitividad. Funding: Duration: CIBERNED Annual Report / 147

148 GROUP Jon Infante Ceberio 601 Principal Investigator Jon Infante Ceberio Research team José Berciario Professor of Neurology and associate professor emeritus María Teresa Berciario Full professor of Cell Biology María Bravo Nurse Iñigo Casafont Assistant professor Marta de la Fuente CIBERNED part-time administrative assistant Antonio García Clinical neurophysiology ASP Elena Gallardo Area specialist physician (ASP) and assistant professor of radiology Isabel González- Aramburu Neurology ASP Andrés González-Mandly Radiology ASP Andrea González Suarez Neurology specialist Marta Kazimierczak Nurse Miguel A. Lafarga Full professor or Cell Biology Carmen Lage Neurology specialist José Ignacio Mateo Neurology ASP Ana Lara Pelayo Neurology ASP Ana Pozueta Psychologist Javier Riancho- Zarrabeitia Neurology specialist Eloy Rodríguez Rodríguez Neurology ASP Pascual Sánchez-Juan Neurology ASP Coro Sánchez- Quintana Lab technician Olga Tapia Martínez Postdoctoral researcher María J. Sedano Neurology ASP María Sierra Neurology ASP Onofre Combarros Pascual Full professor of Neurology Jorge Mata Garrido Servicio de Neurología Hospital Universitario Marqués de Valdecilla Universidad de Cantabria Santander, España 2016 CIBERNED Annual Report / 148

149 Summary Peripheral Neuropathies We have demonstrated that in Guillain-Barré syndrome (GBS), sural nerve inflammatory lesions might exist even when nerve conduction parameters are normal. We have described that the absence of the H-wave in the paraparetic variant of GBS can be the only initial manifestation. In a CMT pedigree associated with NEFL N98S mutation we have demonstrated that mutations in this gene are a new cause of autosomal dominant-intermediate form (DI-CMT). In a pedigree of axonal CMT (CMT2), followed by us for four decades, next generation sequencing has yielded a mutation in the gen LRSAM1, implying pcyst695tyr. The clinico-molecular correlation has involved the performance of consecutive examinations including neurophysiological exams and MRI studies in lower limbs; only by means of this way we have been able to define that the phenotypic expression is age-dependent, with cases of incomplete penetrance. Muscular MRI systematically showed distal predominant fat atrophy, as it corresponds to a lengthdependent axonopathy. Parkinson s disease We have carried out a comparative transcriptomic analysis in peripheral blood between patients with idiopathic PD and those carrying the G2019S mutation of the LRRK2 gene. We have identified different patterns of genetic expression between both forms of PD and we have defined a wide set of genes whose expression might be altered by the presence of the G2019S mutation. On the other side, we have completed the four-year followup of a cohort of asymptomatic carriers of the G2019S mutation of LRRK2, studied by molecular neuroimaging (DaTSCAN) and olfaction testing. We have defined the usefulness of DaTSCAN as a tool for predicting risk of early conversion to PD in the cohort of asymptomatic carriers. Alzheimer s disease Our group is a founder member of the Spanish COnsortium of DEmentia Genetics (DEGESCO). Within this collaborative project we have reported the association between genetic variants of MAPT gene and different neurodegenerative disorders in a sample of subjects. In another initiative of DEGESCO we have studied the role of rare variants in TUBA4A gene in frontotemporal dementia. We have participated in a study of the genomic and neuroimaging consortium ENIGMA, analyzing the brain subcortical asymmetries in a large sample of 15,847 subjects. Hereditary Ataxias During 2016 we have continued the prospective evaluation of patients with dominant ataxias included in the studies EUROSCA-NHS and RISCA. We are a collaborator centre in the european study ESMI (European spinocerebellar ataxia type 3 /Machado Joseph initiative), within JPND 2015 framework; the recruitment period having already started in On the other side, we have participated in a GWAs study of patients with multiple system atrophy and have contributed to define the participation of the peripheral nerves in the most prevalent forms of dominant ataxias in an international study. Amyotrophic lateral sclerosis (ALS) We have reported that the incidence of ALS in Cantabria has almost doubled in the last 40 years. Also, we have shown a North-South gradient for mortality caused by ALS in Spain and reported an inverse association with levels of gross domestic product. In the line of treatments, we have published several papers supporting the role of retinoids as a new treatment for ALS. Cell neurobiology Our work has focused in three research topics: 1) 2) The regulatory effect of acetylation of the survival motor neuron (SMN) protein on the biogenesis of spliceosomal snrnps and on the molecular assembly of Cajal Bodies (CBs). The contribution of the CB and the nucleolus to the molecular pathophysiology of the spinal muscular atrophy (SMA) in motor neurons. 3) DNA damage and repair in neurons. Regarding the first point, we have demonstrated that both endogenous and overexpressed SMN are acetylated in the lysine K119 by the acetiltranferase CBP. This modification changes SMN interactome and functions resulting in reduced spliceosomal snrnp biogenesis and deficient CB assembly. Concerning the second point, in a murine model of SMA type I we have demonstrated in motor neurons that depletion of CBs and the associated alterations in nucleolar transcription machinery are nuclear hallmarks of the dysfunction of RNA metabolism in this disease. Concerning the third point, we have demonstrated with ChIP-seq analysis that the nuclear foci of unrepaired DNA in cerebral cortical neurons are enriched in specific DNA sequences, including 17 high vulnerability genes encoding proteins related with human neuromuscular disorders CIBERNED Annual Report / 149

150 Keywords Guillain Barre syndrome, ataxia, Charcot-Marie-Tooth disease, disease of Parkinson, Alzheimer disease, Creutzfeldt-Jakob disease, cajal nuclear bodies, amyotrophic lateral sclerosis Publications Peeters K., Palaima P., Pelayo-Negro A.L., Garcia A., Gallardo E., Garcia-Barredo R. et al. Charcot-Marie-Tooth disease type 2G redefined by a novel mutation in LRSAM1. Annals of Neurology Talamillo A., Grande L., Ruiz-Ontanon P., Velasquez C., Mollinedo P., Torices S. et al. ODZ1 allows glioblastoma to sustain invasiveness through a Myc-dependent transcriptional upregulation of RhoA. Oncogene Sailer A., Scholz S.W., Nalls M.A., Schulte C., Federoff M., Price T.R. et al. A genome-wide association study in multiple system atrophy. Neurology. 2016;87(15): Riancho J. Retinoids and PPAR agonists: Promising partners in neurodegenerative diseases?. Free Radical Biology and Medicine Lafita-Navarro MC, Blanco R, Mata-Garrido J, Liaño-Pons J, Tapia O, García-Gutiérrez L et al. MXD1 localizes in the nucleolus, binds UBF and impairs rrna synthesis.oncotarget Lafarga M, Tapia O, Romero AM, Berciano MT. Cajal bodies in neurons.rna biology Sierra M., Gelpi E., Marti M.J., Compta Y. Lewy- and Alzheimer-type pathologies in midbrain and cerebellum across the Lewy body disorders spectrum. Neuropathology and Applied Neurobiology. 2016;42(5): Jimenez-Bonilla J.F., Banzo I., De Arcocha-Torres M., Quirce R., Martinez-Rodriguez I., Sanchez- Juan P. et al. Amyloid imaging with 11C-PIB in patients with cognitive impairment in a clinical setting: A visual and semiquantitative analysis. Clinical Nuclear Medicine. 2016;41(1):e18-e23. Guadalupe T., Mathias S.R., VanErp T.G.M., Whelan C.D., Zwiers M.P., Abe Y. et al. Human subcortical brain asymmetries in 15,847 people worldwide reveal effects of age and sex. Brain Imaging and Behavior. 2016; Riancho J., Berciano M.T., Berciano J., Lafarga M. Relaunching an old drug: the potential role of bexarotene in neurodegenerative diseases. Journal of Neurology. 2016; 1-2. Berciano J., Sedano M.J., Pelayo-Negro A.L., Garcia A., Orizaola P., Gallardo E. et al. Proximal nerve lesions in early Guillain Barré syndrome: implications for pathogenesis and disease classification. Journal of Neurology. 2016; Riancho J., Delgado-Alvarado M., Fernandez-Torre J.L., Sanchez-Juan P., Polo J.M. Subacute progressive aphasia: a rare presentation of Creutzfeldt Jakob disease. Journal of Neurology. 2016; 1-3. Riancho J., Lozano-Cuesta P., Santurtun A., Sanchez-Juan P., Lopez-Vega J.M., Berciano J. et al. Amyotrophic Lateral Sclerosis in Northern Spain 40 Years Later: What Has Changed. Neurodegenerative Diseases. 2016; Berciano J., Peeters K., Garcia A., Lopez-Alburquerque T., Gallardo E., Hernandez-Fabian A. et al. NEFL N98S mutation: another cause of dominant intermediate Charcot Marie Tooth disease with heterogeneous early-onset phenotype. Journal of Neurology. 2016;263(2): Salas-Gomez D, Fernandez-Gorgojo M, Pozueta A, Diaz-Ceballos I, Lamarain M, Perez C et al. Binge Drinking in Young University Students Is Associated with Alterations in Executive Functions Related to Their Starting Age.PloS one. 2016;11(11). Martín-Láez R, Valle-San Román N, Rodríguez-Rodríguez EM, Marco-de Lucas E, Berciano Blanco JA, Vázquez-Barquero A. Current concepts on the pathophysiology of idiopathic chronic adult hydrocephalus: Are we facing another neurodegenerative disease?. Neurologia (Barcelona, Spain) CIBERNED Annual Report / 150

151 Program 2 Group: Jon Infante Ceberio Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Santurtun A., Villar A., Delgado-Alvarado M., Riancho J. Trends in motor neuron disease: association with latitude and air lead levels in Spain. Neurological Sciences. 2016; 1-5. Santurtún A, Delgado-Alvarado M, Villar A, Riancho J. [Geographical distribution of mortality by Parkinson s disease and its association with air lead levels in Spain].Medicina clinica Jiménez-Bonilla JF, Banzo I, De Arcocha-Torres M, Quirce R, Martínez-Rodríguez I, Lavado- Pérez C et al. Diagnostic role of 11C-Pittsburgh compound B retention patterns and glucose metabolism by fluorine-18-fluorodeoxyglucose PET/CT in amnestic and nonamnestic mild cognitive impairment patients.nuclear medicine communications. 2016;37(11). Berciano J, Gallardo E, Orizaola P, Marco de Lucas E, García A, Pelayo-Negro AL et al. Comment on Paraparetic Guillain-Barré syndrome: non-demyelinating reversible conduction failure restricted to the lower limbs.muscle & nerve Fernandez-Fernandez J, Real-Noval H, Rodriguez-Rodriguez E. [Massive cerebral air embolism following endoscopic retrograde cholangiopancreatography. A case report and review of the literature].revista de neurologia. 2016;63(11). Velasquez C, Caballero H, Bucheli C, Berciano J, Vazquez-Barquero A, Martino J. [A very slowgrowing exophytic hemisphere glioma: a case report].revista de neurologia. 2016;62(1). Riancho J, Del Real A, Riancho JA. How to interpret epigenetic association studies: a guide for clinicians.bonekey reports. 2016;5. Mata-Garrido J, Casafont I, Tapia O, Berciano MT, Lafarga M. Neuronal accumulation of unrepaired DNA in a novel specific chromatin domain: structural, molecular and transcriptional characterization.acta neuropathologica communications. 2016;4. Berciano J., Gallardo E., Orizaola P., De Lucas E.M., Garcia A., Pelayo-Negro A.L. et al. Early axonal Guillain-Barré syndrome with normal peripheral conduction: Imaging evidence for changes in proximal nerve segments. Journal of Neurology, Neurosurgery and Psychiatry. 2016;87(5): Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. Santurtun A., Villar A., Lopez-Delgado L., Riancho J. Amyotrophic lateral sclerosis and richness: A correlation study across Spain. Journal of the Neurological Sciences. 2016;367: Riancho J., Navasa J.M., Sedano-Tous M.J., Polo J.M. Carotid-cavernous fistula successfully treated with carotid compression. Medicina Clinica. 2016;146(1):48-. Riancho J, Jiménez-López Y, Sánchez-de la Torre JR, Pardina-Vilella L, Sánchez-Juan P. How much do Spanish clinicians know about dementia?. Journal of the neurological sciences Research projects Code: PI2016/06. Title: Identificación de vías fisiopatológicas y biomarcadores candidatos en la fase prediagnóstica de la enfermedad de Parkinson. Principal Investigator: Miquel Vila Bover. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G601, G207, G410. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 151

152 Code: NVAL 2015/17. Title: Estudio de la utilidad de biomarcadores de neuroimagen y olfación en la enfermedad de Parkinson asociada a la mutación G2019S de LRRK2 como predictores de conversión a enfermedad de Parkinson motora. Principal Investigator: María Sierra Pena. CIBERNED groups: G601. Type: National. Funding agency: IDIVAL. Funding: Duration: Code: PI11/ Title: Selección de genes candidatos para la enfermedad de Parkinson idiopática a traves del análisis diferencial del transcriptoma en pacientes y portadores asintomaticos de mutaciones en LRRK2. Principal Investigator: Jon Infante Ceberio. CIBERNED groups: G601. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: ,55. Duration: Code: CINERNED 2014/02. Title: Mecanismos epigenéticos implicados en la etiología y progresión de las demencias neurodegenerativas rapidamente progresivas. Principal Investigator: Miguel Calero. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G601. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: PI13/ Title: Biomarcadores de enfermedad de Alzheimer como factores pronóstico en hidrocefalia a presión normal idiomática. Principal Investigator: Eloy Rodríguez. CIBERNED groups: G601. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: ,5. Duration: Code: PI12/ Title: Estudio multimodal de biomarcadores de enfermedad de Alzheimer en deterioro cognitivo postoperatorio. Principal Investigator: Pascual Sánchez- Juan. CIBERNED groups: G601. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: SMA Europe Call 8 grant application. Title: Regulation of the survival motor neuron (SMN) protein by acetylation and its importance in snrnp biogenesis and molecular assembly of Cajal bodies. Principal Investigator: Olga Tapia Martínez. CIBERNED groups: G601. Type: European. Funding agency: Comisión Europea. Funding: N.D. Duration: PhD dissertations Author: María Ruiz Soto. Title: El estrés oxidativo en la glía satélite de los ganglios raquídeos induce alteraciones sensitivas en el modelo murino hsod1(g93a) de esclerosis lateral amiotrófica (ELA). Date: 5 February Supervisor: Miguel Ángel Lafarga Coscojuela. Author: Isabel González Aramburu. Title: Niveles séricos de ácido úrico y progranulina, factores genéticos que los regulan y enfermedad de Parkinson. Date: 5 February Supervisor: Jon Infante Ceberio CIBERNED Annual Report / 152

153 GROUP Jaime Kulisevsky Bojarski 202 Principal Investigator Jaime Kulisevsky Bojarsky Research team Helena Berj Kasem Marco Researcher Antonia Campolongo Perillo Researcher Carmen García Sánchez Researcher Alexandre Gironell Carreró Researcher Javier Pagonabarraga Mora Researcher Berta Pascual Sedano Researcher María Victoria Sosti Sosa Researcher Juan Martín Lahoz Saúl Martínez Horta Jesús Pérez Pérez Sara Belmonte Calderón Researcher Ignacio Aracil Bolaños Ramón Fernández de Bobadilla Researcher Carles Roig Arnall Researcher Unidad de trastornos de movimiento, Servicio de Neurología Hospital de la Santa Creu i Sant Pau Sant Antoni M. Claret Barcelona, Catalunya (Spain) Tel.: Fax: jkulisevsky@santpau.cat 2016 CIBERNED Annual Report / 153

154 Summary Directed by Jaime Kulisevsky MD, PhD (Movement Disorders Unit, Sant Pau Hospital, Biomedical Research Institute), we focus on research of Parkinson s Disease (PD) and other Movement Disorders- During 2016 we initiated new projects and achieved different objectives: Genetics Parkinson and PSP: Study of genetic variations and risk of development of neurodegenerative diseases (2016). Parkinson and cognition FIS PI14/02058: Blood markers and neurophysiological progression of cognitive impairment. Validation of cognitive (PD-CRS) and functional scales (PD-CFRS) with diagnostic criteria of mild cognitive impairment (MCI) in Parkinson and other neurodegenerative diseases (published in J Neur Sci). Development of alternative version of PD- CRS for reevaluations in clinical trials (article under review). Validation and normative data of PD-CRS by research groups from other countries. Dissemination, translation and validation of neuropsychological scales designed for Parkinson in a international web platform ( DuoCog project: randomized, double-blind and cross-over study comparing immediate vs- intraduodenal levodopa in cognition and humor (own study, with funding from Abbvie International). Intramural project with Ciberned support: A randomized, double-blind, placebocontrolled clinical trial evaluating the efficacy and safety of candesartan versus placebo in mild cognitive impairment associated with Parkinson s disease. Apathy and behavior in parkinson FIS PI15/00962: Anatomical-functional correlates of structural abnormalities of apathy, hallucinations and cognitive impairment. Marató TV3 project: Prediction of impulse control disorders (ICD) and apathy- Anatomical-functional correlates of ICD and apathy. Advanced parkinson Continues the analysis of the nutritional status in patients with intraduodenal levodopa. Prospective study of cognitive, behavioral, sleep and quality of life aspects in patients undergoing deep brain stimulation. Utility study of a remote motor activity measurement system (PKG, Global Kinetics Corporation). Huntington s disease (HD) Coordination and development of cognitive assessment instruments of the Cognitive Phenotype Working Group of the European Huntington s Disease Network (EHDN). Spanish Coordination of the study Enroll- HD: A prospective registry study in a global Huntington s Disease cohort. Collaboration with the UB in the projects The nuclear lamina in EH: pathophysiology and therapeutic applications (Ramón Areces Grant) and Overcoming Depression in HD: Cdk5 as a potential biomarker and pharmacological target (Huntington s Disease Society of America). Collaboration in FIS PI14/00834: Longitudinal changes in functional and structural connectivity in pre-symptomatic patients and FIS PI15/02227: Phenotype and haplotypes associated with intermediate alleles of the HD gene. Tremor Development of hardware-software system of portable neurophysiological study of movement disorders- Collaborative project with the company SEIDOR and the University of Vic. Study of typification of tremor syndromes in Parkinson. Dystonia Spanish consensus on Evidence and myths about the use of botulinum toxin in dystonia and spasticity (paper accepted for publication). Ataxia-premutation tremor associated with fragile x syndrome (FXTAS) Assistance and collaborative research with Genetics of the Hospital Clínic (Barcelona) and with the Catalan Association of X Fràgil (ACXF). Detection of clinical and technological variables predictive of FXTAS progression in patients with premutation of Fragile X cromosome. Translation research Huntington transgenic model in rat (BACHD): longitudinal study of motor, emotional and cognitive alterations. Rat hemiparkinsonism model: study of estriatal transcriptional changes with different doses of L-Dopa CIBERNED Annual Report / 154

155 Program 2 Group: Jaime Kulisevsky Bojarski Keywords Cognition, mild cognitive impairment, huntington, dystonia, FxTAS, essential tremor, animal models Publications Schapira AH, Fox SH, Hauser RA, Jankovic J, Jost WH, Kenney C et al. Assessment of Safety and Efficacy of Safinamide as a Levodopa Adjunct in Patients With Parkinson Disease and Motor Fluctuations: A Randomized Clinical Trial.JAMA neurology. 2017;74(2). Epub Rodriguez-Revenga L., Pagonabarraga J., Gomez-Anson B., Lopez-Mourelo O., Izquierdo S., Alvarez-Mora M.I. et al. Carriage of One or Two FMR1 Premutation Alleles Seems to Have No Effect on Illness Severity in a FXTAS Female with an Autozygous FMR1 Premutation Allele. Cerebellum. 2016; 1-8. Spataro N., Roca-Umbert A., Cervera-Carles L., Valles M., Anglada R., Pagonabarraga J. et al. Detection of genomic rearrangements from targeted resequencing data in Parkinson s disease patients. Movement Disorders López-Mourelo O, Mur E, Madrigal I, Alvarez-Mora MI, Gómez-Ansón B, Pagonabarraga J et al. Social anxiety and autism spectrum traits among adult FMR1 premutation carriers.clinical genetics Martinez-Horta S., Sampedro F., Pagonabarraga J., Fernandez-Bobadilla R., Marin-Lahoz J., Riba J. et al. Non-demented Parkinson s disease patients with apathy show decreased grey matter volume in key executive and reward-related nodes. Brain Imaging and Behavior. 2016; 1-9. Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. Crespi J., Brathen G., Quist-Paulsen P., Pagonabarraga J., Roig-Arnall C. Facial Dystonia with Facial Grimacing and Vertical Gaze Palsy with round the Houses Sign in a 29-Year-Old Woman. Neuro-Ophthalmology. 2016;40(1): Provenzano C, Zamboni M, Veneziano L, Mantuano E, Garavaglia B, Zorzi G et al. Functional characterization of two novel mutations in TTF-1/NKX2.1 homeodomain in patients with benign hereditary chorea.journal of the neurological sciences. 2016;360. Matias-Guiu J.A., Fernandez-Bobadilla R., Fernandez-Oliveira A., Valles-Salgado M., Rognoni T., Cortes-Martinez A. et al. Normative data for the Spanish version of the addenbrooke s cognitive examination III. Dementia and Geriatric Cognitive Disorders. 2016;41(5-6): Lopez-Mora D.A., Camacho V., Perez-Perez J., Martinez-Horta S., Fernandez A., Sampedro F. et al. Striatal hypometabolism in premanifest and manifest Huntington s disease patients. European Journal of Nuclear Medicine and Molecular Imaging. 2016;43(12): Martinez-Horta S, Perez-Perez J, van Duijn E, Fernandez-Bobadilla R, Carceller M, Pagonabarraga J et al. Corrigendum to Neuropsychiatric symptoms are very common in premanifest and early stage Huntington s disease [Parkinsonism Relat. Disord. 25C (2016) 58-64].Parkinsonism & related disorders. 2016;31. Martinez-Horta S., Perez-Perez J., van Duijn E., Fernandez-Bobadilla R., Carceller M., Pagonabarraga J. et al. Erratum: Corrigendum to Neuropsychiatric symptoms are very common in premanifest and early stage Huntington s disease (Parkinsonism and Related Disorders (2016) 25C (58 64) (S ) ( /j.parkreldis )). Parkinsonism and Related Disorders. 2016;31:161-. Gironell A. Routine neurophysiology testing and functional tremor: Toward the establishment of diagnostic criteria. Movement Disorders. 2016;31(11): Gironell A., Marin-Lahoz J. The essence of essential tremor: Neurochemical bases. Revista de Neurologia. 2016;62(11): CIBERNED Annual Report / 155

156 Research projects Code: PI2015/02. Title: Validación de nuevas dianas terapéuticas y nuevos biomarcadores en enfermedad de Parkinson. Principal Investigator: José Luis Labandeira García. CIBERNED s collaboration: Yes. CIBERNED groups: G208, G101, G202, G203. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: MARATO Title: Blood-based and neurophysiological markers of cognitive deterioration and dementia in Parkinson s disease. Principal Investigator: Javier Pagonabarraga. CIBERNED groups: G202. Type: Private. Funding agency: Fundació La Marató de TV3. Funding: ,25. Duration: Code: 477/U/2014 MARATO. Title: Prediction of apathy and impulse control disorders in Parkinson Disease based on feedback related negativity. Principal Investigator: Jaime Kulisevsky. CIBERNED groups: G202. Type: Private. Funding agency: Fundació La Marató de TV3. Funding: ,5. Duration: Code: PI15/ Title: Correlatos anatómico-funcionales de los trastornos del control de impulsos y apatía en la enfermedad de Parkinson. Principal Investigator: Jaime Kulisevsky. CIBERNED groups: G202. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: 2010 SGR Title: AGAUR SGR Principal Investigator: Jaime Kulisevsky. CIBERNED groups: G202. Type: CC.AA. Funding agency: AGAUR-Generalitat de Catalunya Funding: Duration: Code: CM15/ Title: Río Hortega Principal Investigator: Juan Marín Lahoz. CIBERNED groups: G202. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: CIBERNED Annual Report / 156

157 GROUP José Luis Labandeira García 208 Principal Investigator José Luis Labandeira García Full professor Research team María 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 Associate professor Begoña Villar Cheda Associate professor Pablo Garrido Gil Postdoctoral researcher Rita Valenzuela Limiñana Predoctoral researcher Juan A. Parga Martín Postdoctoral researcher Antonio Domínguez Meijide Postdoctoral researcher Mª Alicia Costa Besada Predoctoral researcher María García Garrote Predoctoral researcher Iria Novoa Pérez Specialist technician Center for Research in Molecular Medicine and Chronic Diseases (CIMUS) Universidad de Santiago de Compostela Avenida Barcelona, 22 Santiago de Compostela (Spain) Tel.: / Fax: Pilar Aldrey García Specialist technician José A. Trillo Franco Specialist technician Mª Cristina Gianzo Quintela Specialist technician 2016 CIBERNED Annual Report / 157

158 Summary In 2016, we investigated mechanisms involved in higher vulnerability of dopaminergic neurons with aging and mechanisms involved in progression of Parkinson s disease (PD). We collaborated with other Ciberned research groups to develop new animal models of PD (J Neurochem. 136: ). We also investigated the effects of estrogen, menopause and hormonal therapies in development and progression of PD (Front. Neurendoc rinol 43:44-59). The role of Rho kinase (ROCK) in agerelated increase in dopaminergic vulnerability (J Gerontol A Biol Sci Med Sci. 71:1254-7), and survival of dopaminergic cells grafted in PD animal models (Stem Cells Trasl Med 5: ) was also investigated. Other mechanisms involved in brain aging and dopaminergic vulnerability, and their possible manipulation for neuroprotection in PD were investigated in several studies published in We described for the first time angiotensin receptors in brain mitochondria and their possible role in neurodegeneration and aging (Cell Death Dis 7: e2427). The role of brain IGF-1 in aging, neuroinflammation and dopaminergic degeneration, and particularly the interactions between IGF-1 and brain RAS were analyzed in other study (Oncotarget 7: ). In collaboration with different Ciberned research groups, we further investigated the role of striatal D1 and D2 dopaminergic receptors in progression of PD and L-dopa-induced dyskinesias in rodents and primates (Mol Neurobiol 53: ; Brain Struct Func PMID ). In addition we collaborated in the study of new IMAO compounds (multitarget inhibitors of MAO B and Cholinesterases) as possible new antiparkinsonian drugs (J Med Chem 59: ; Molecules 21: 362). An important part of the above-mentioned studies have been performed in collaboration with different Ciberned research groups : Dr. Lanciego, Drs Canela/R. Franco, Dr. López-Barneo/Toledo. Keywords Neurodegeneration, neuroprotection, neuroinflammation, Parkinson, aging, angiotensin, cell therapy Publications Rico A.J., Dopeso-Reyes I.G., Martinez-Pinilla E., Sucunza D., Pignataro D., Roda E. et al. Neurochemical evidence supporting dopamine D1 D2 receptor heteromers in the striatum of the long-tailed macaque: changes following dopaminergic manipulation. Brain Structure and Function. 2016; Iglesias-Gonzalez J., Sanchez-Iglesias S., Beiras-Iglesias A., Mendez-Alvarez E., Soto-Otero R. Effects of Aluminium on Rat Brain Mitochondria Bioenergetics: an In vitro and In vivo Study. Molecular Neurobiology. 2016; 1-8. Labandeira-Garcia JL, Rodriguez-Perez AI, Valenzuela R, Costa-Besada MA, Guerra MJ. Menopause and Parkinson s disease. Interaction between estrogens and brain reninangiotensin system in dopaminergic degeneration.frontiers in neuroendocrinology Pisani L., Farina R., Catto M., Iacobazzi R.M., Nicolotti O., Cellamare S. et al. Exploring Basic Tail Modifications of Coumarin-Based Dual Acetylcholinesterase-Monoamine Oxidase B Inhibitors: Identification of Water-Soluble, Brain-Permeant Neuroprotective Multitarget Agents. Journal of Medicinal Chemistry. 2016;59(14): Pisani L., Farina R., Soto-Otero R., Denora N., Mangiatordi G.F., Nicolotti O. et al. Searching for multitargeting neurotherapeutics against Alzheimer s: Discovery of potent AChE-MAO B inhibitors through the decoration of the 2H-Chromen-2-one structural motif. Molecules. 2016;21(3). Rodriguez-Perez A.I., Borrajo A., Diaz-Ruiz C., Garrido-Gil P., Labandeira-Garcia J.L. Crosstalk between insulin-like growth factor-1 and angiotensin-ii in dopaminergic neurons and glial cells: Role in neuroinflammation and aging. Oncotarget. 2016;7(21): Rodriguez-Pallares J., Rodriguez-Perez A.I., Munoz A., Parga J.A., Toledo-Aral J.J., Labandeira- Garcia J.L. Effects of rho kinase inhibitors on grafts of dopaminergic cell precursors in a rat model of Parkinson s disease. Stem Cells Translational Medicine. 2016;5(6): CIBERNED Annual Report / 158

159 Program 2 Group: José Luis Labandeira García Valenzuela R, Costa-Besada MA, Iglesias-Gonzalez J, Perez-Costas E, Villar-Cheda B, Garrido- Gil P et al. Mitochondrial angiotensin receptors in dopaminergic neurons. Role in cell protection and aging-related vulnerability to neurodegeneration.cell death & disease. 2016;7(10). Research projects Code: PI 2015/02. Title: Validación de nuevas dianas terapéuticas y nuevos biomarcadores en enfermedad de Parkinson. Principal Investigator: José Luis Labandeira. CIBERNED s collaboration: Yes. CIBERNED groups: G208, G101, G202, G203. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: RD12/0019/0008. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas Coordinador de red: José María Moraleda. CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RD16/0011/0017. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas. CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: GRC2014/002. Title: Consolidación y estructuración de grupos de referencia competitiva. Prncipal Investigator: José Luis Labandeira García. CIBERNED groups: G208. Type: CC.AA. Funding agency: Xunta Galicia. Funding: Duration: Code: 2015-CE009. Title: Financiación línea de investigación en enf. neuromuscuylares. Trastornos del movimiento. Principal Investigator: José luis Labandeira. CIBERNED groups: G208. Type: Private. Funding agency: Fundación de La Luz. Funding: Duration: Code: BFU Title: Sistema renina-angiotensina cerebral en neuroinflamación y degeneración dopaminérgica. Más allá del eje AII/AT1/ NADPH-oxidase. Principal Investigator: José Luis Labandeira. CIBERNED groups: G208. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: PhD dissertations Author: Ana Borrajo López. Title: Mecanismos de activación microglial en degeneración dopaminérgica: papel del eje Angiotensina/AT1/ Rho Quinasa. Date: 15 January Supervisor: José Luis Labandeira García CIBERNED Annual Report / 159

160 GROUP José Luis Lanciego Pérez 203 Principal Investigator José Luis Lanciego Research team Gloria González-Aseguinolaza Researcher Alberto José Rico Martín Researcher Iria María González-Dopeso Reyes Researcher José Diego Pignataro Diego Sucunza Guibert Elvira Roda Recalde Lab technician Fundación para la Investigación Médica Aplicada (FIMA) Centro de Investigación Médica Aplicada (CIMA) Departamento de Neurociencias Laboratorio de Neuroanatomía de Ganglios Basales Avenida Pío XII, Edificio CIMA Pamplona, Navarra (Spain) Tel.: (2002) Fax: CIBERNED Annual Report / 160

161 Summary Studies on heteromers made of G proteincoupled receptors (GPCRs). It might be argued that GPCRs have a natural tendency to form heteromeric complexes that represent novel molecular entities with properties distinct from those of each component receptor, when considered separately. GPCR heteromers represent novel targets for pharmacological developments in the field of Parkinson s disease. At present, our main activities focus on GPCR heteromers made of dopaminergic receptors (D1 and D2), as well as those formed by endocannabinoid receptors (CB1, CB2 and GPR55). Gene therapy for Parkinson s disease. The field of gene therapy has recently witnessed a numer of major conceptual changes. Instead of using viral vectors for the delivery of a given therapeutic gene, the current tendency in the field relies on the use of gene-carrying viral vectors for modifying brain circuits. It is expected that these genetic manipulations of brain circuits will ultimately sustain a direct therapeutic effect. By using relevant animal models of Parkinson s disease, we have designed different types of adeno-associated (AAVs) and lentiviral vectors that are currently being explored within the following specific projects: Direct in situ cellular reprogramming using AAVs coding for a combination of 3 transcription factors. Selective clearance of alpha-synuclein aggregates by means of pseudotyped lentiviruses carrying the gene coding for beta-glucocerebrosidase. Keywords GPCR, dopamine, cannabis, basal ganglia, gene therapy, adeno-associated viral vectors, lentiviruses Publications Rico A.J., Dopeso-Reyes I.G., Martinez-Pinilla E., Sucunza D., Pignataro D., Roda E. et al. Neurochemical evidence supporting dopamine D1 D2 receptor heteromers in the striatum of the long-tailed macaque: changes following dopaminergic manipulation. Brain Structure and Function. 2016; Franco R., Martinez-Pinilla E., Lanciego J.L., Navarro G. Basic pharmacological and structural evidence for class A G-protein-coupled receptor heteromerization. Frontiers in Pharmacology. 2016;7. Garbayo E., Ansorena E., Lana H., Carmona-Abellan M.D.M., Marcilla I., Lanciego J.L. et al. Brain delivery of microencapsulated GDNF induces functional and structural recovery in parkinsonian monkeys. Biomaterials. 2016;110: Perreault M.L., Hasbi A., Shen M.Y.F., Fan T., Navarro G., Fletcher P.J. et al. Disruption of a dopamine receptor complex amplifies the actions of cocaine. European Neuropsychopharmacology. 2016;26(9): Valenzuela R, Costa-Besada MA, Iglesias-Gonzalez J, Perez-Costas E, Villar-Cheda B, Garrido- Gil P et al. Mitochondrial angiotensin receptors in dopaminergic neurons. Role in cell protection and aging-related vulnerability to neurodegeneration.cell death & disease. 2016;7(10). Martinez-Pinilla E., Rabal O., Reyes-Resina I., Zamarbide M., Navarro G., Sanchez-Arias J.A. et al. Two affinity sites of the cannabinoid subtype 2 receptor identified by a novel homogeneous binding assay. Journal of Pharmacology and Experimental Therapeutics. 2016;358(3): CIBERNED Annual Report / 161

162 Research projects Code: PI 2015/02. Title: Validación de nuevas dianas terapéuticas y nuevos biomarcadores en enfermedad de Parkinson. Principal Investigator: José Luis Labandeira. CIBERNED s collaboration: Yes. CIBERNED groups: G208, G101, G202, G203. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: Title: Cannabinoid receptor heteromeric complexes as therapeutic targets in Parkinson s disease. Principal Investigator: Rafael Franco Fernández. CIBERNED s collaboration: Yes. CIBERNED groups: G201, G203. Type: Private. Funding agency: Fundació La Marató de TV3. 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). CIBERNED groups: G203. Type: International. Funding agency: European Research Council Executive Agency Funding: Duration: CIBERNED Annual Report / 162

163 GROUP José López Barneo 105 Principal Investigator José López Barneo Research team Candela Caballero Erazo Doctor Ivette López López Technician Patricia González Rodríguez Researcher Francisco Javier Villadiego Luque Postdoctoral researcher Gloria Paula García Flores Technician Helia Sarmiento Soto Technician Ignacio Arias Mayenco Technician Juan José Toledo Aral Researcher Lin Gao Researcher Nela Suárez Luna Technician Pablo Mir Rivera Doctor Instituto de Biomedicina de Sevilla Hospital Universitario Virgen del Rocío CISC, Universidad de Sevilla Avenida Manuel Siurot s/n Sevilla (Spain) Tel.: Fax: Patricia Ortega Sáenz Researcher Pilar Gómez Garre Researcher Ricardo Pardal Redondo Researcher Victoria Bonilla Henao Technician Xavier d Anglemont de Tassigny Postdoctoral researcher 2016 CIBERNED Annual Report / 163

164 Summary In 2016, our group has developed three research lines related to CIBERNED: Pathogenesis of Parkinson s disease (PD): We are currently studying the early stages of neurodegenerative diseases, such as Parkinson s disease (PD), using two transgenic animal models carrying deletions in mitochondrial complexes I and III. Part o this work is done in collaboration with the laboratory of Dr. James Surmeier of Northwestern University (Chicago, USA). We have developed several animal models that are now in the characterization step to i) advance in the understanding of the role of endogenous neurotrophic glial derived factor (GDNF) on the maintenance of the nigrostriatal pathway, ii) study the molecular mechanisms leading to the selective synthesis and secretion of GDNF by parvalbumin positive (PV+) interneurons in the striatum, and iii) investigate methods to increase the intrastriatal release of GDNF. For this, we have developed a sorting protocol to compare genomic and proteomic patterns of the striatal (GDNF+) and cortical (GDNF-) PV+ neurons. This will help to understand the specific regulation of endogenous GDNF synthesis. In addition, we have initiated a collaborative project with the labroatory of Dr. José Obeso to characterize PV+ interneurons in the monkey striatum and the effects of chronic administration of MPTP. 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: We continue our collaboration with Dr. Juan José Toledo-Aral s group on trofic effects of carotid body (CB) transplants in models of PD. We have analyzed several factors (i.e. age and sex of the human donors) that may influence the action exerted by the transplanted CB tissue. In this regard, age is 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. Effect of hypoxia on the neurogenic niches in the central and peripheral nervous system: We continue with the investigation on the effects of chronic and intermittent hypoxia on the central (brain) and peripheral (CB) neurogenic niches. In collaboration with Dr Ricardo Pardal s group we have identified a new cell type (neuroblasts) in the adult CB, which has a fundamental role in the fast generation of new O2-sensitive neuron-like chemoreceptor cells. We are working on the elucidation of the role of these neurobalsts in human pathologies with CB overactivation and an excess of sympathetic outflow. Role of biotin in dopaminergic systems. We have found that the CB contains unusual high levels of biotin and that this vitamin/ coenzyme is necessary for the uptake and maturation of dopaminergic secretory vesicles. The role of biotin on central dopaminergic transmission is under study. Keywords Hypoxia, ion channels, carotid body, movement disorders, Parkinson s disease Publications Lohmann K., Schlicht F., Svetel M., Hinrichs F., Zittel S., Graf J. et al. The role of mutations in COL6A3 in isolated dystonia. Journal of Neurology. 2016; 1-5. Navarro-Guerrero E., Platero-Luengo A., Linares-Clemente P., Cases I., Lopez-Barneo J., Pardal R. Gene Expression Profiling Supports the Neural Crest Origin of Adult Rodent Carotid Body Stem Cells and Identifies CD10 as a Marker for Mesectoderm-Committed Progenitors. Stem Cells Pardal R., Lopez Barneo J. Mature neurons modulate neurogenesis through chemical signals acting on neural stem cells. Development Growth and Differentiation Lopez-Barneo J, Gonzalez-Rodriguez P, Gao L, Fernandez-Aguera MC, Pardal R, Ortega- Saenz P. Oxygen sensing by the carotid body: mechanisms and role in adaptation to hypoxia. American journal of physiology. Cell physiology CIBERNED Annual Report / 164

165 Program 2 Group: José López Barneo Gomar I.S., Sanchez M.D., Sanchez A.J.U., Chocan J., Suarez-Luna N., Ramirez-Lorca R. et al. Comparative analysis for the presence of IgG anti-aquaporin-1 in patients with NMO-spectrum disorders. International Journal of Molecular Sciences. 2016;17(8). Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Rodriguez-Pallares J., Rodriguez-Perez A.I., Munoz A., Parga J.A., Toledo-Aral J.J., Labandeira- Garcia J.L. Effects of rho kinase inhibitors on grafts of dopaminergic cell precursors in a rat model of Parkinson s disease. Stem Cells Translational Medicine. 2016;5(6): Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. Valldeoriola F, Grandas F, Santos-García D, Regidor I, Catalán MJ, Arbelo JM et al. Longterm effectiveness of levodopa-carbidopa intestinal gel in 177 Spanish patients with advanced Parkinson s disease.neurodegenerative disease management. 2016;6(4). Ortega-Saenz P., Macias D., Levitsky K.L., Rodriguez-Gomez J.A., Gonzalez-Rodriguez P., Bonilla- Henao V. et al. Selective accumulation of biotin in arterial chemoreceptors: Requirement for carotid body exocytotic dopamine secretion. Journal of Physiology Ruiz-Veguilla M., Martin-Rodriguez J.F., Palomar F.J., Porcacchia P., Alvarez de Toledo P., Perona-Garcelan S. et al. Trait- and state-dependent cortical inhibitory deficits in bipolar disorder. Bipolar Disorders Pouyssegur J., Lopez-Barneo J. Hypoxia in health and disease. Molecular Aspects of Medicine. 2016;47-48:1-2. Enterria-Morales D., Lopez-Lopez I., Lopez-Barneo J., De Tassigny X.D. Striatal GDNF production is independent to circulating estradiol level despite pan- neuronal activation in the female mouse. PLoS ONE. 2016;11(10). Research projects Code: FIS 13 (PI13/01461). Title: Plasticidad neuronal en las discinesias inducidas por levodopa en la enfermedad de Parkinson. Principal Investigator: Pablo Mir Rivera. CIBERNED groups: G105. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: PIE 2012 Excelencia (P12-CTS-2739 MO). Title: Terapia celular en la enfermedad de Parkinson. Principal Investigator: Juan José Toledo Aral. CIBERNED groups: G105. Type: CC.AA. Funding agency: Junta de Andalucía. Funding: Duration: Code: Retos-Colaboracion 15. Title: Desarrollo de una terapia para el tratamiento de variantes genéticas de alfasinucleina en la enfermedad de Parkinson. Principal Investigator: Pablo Mir Rivera. CIBERNED groups: G105. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: Fundación Mutua Madrilena 13. Title: Estudio de asociación de los genes relacionados con plasticidad neuronal y distonia primaria. Principal Investigator: Pablo Mir Rivera. CIBERNED groups: Type: National. Funding agency: Fundación Mutua Madrilena. Funding: Duration: CIBERNED Annual Report / 165

166 Code: Retos-Colaboracion Title: Desarrollo de una terapia para el tratamiento de variantes genéticas de alfasinucleina en la enfermedad de Parkinson. Principal Investigator: Juan José Toledo Aral. CIBERNED groups: G105. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: ,47. Duration: Code: PIE 2011 Excelencia (P11-CTS-7685). Title: Estudio mediante estimulación magnética transcraneal del papel del circuito cerebelo-talamo-cortical en el desarrollo de discinesias en la enfermedad de Parkinson y su modulacion por factores genéticos. Principal Investigator: Pablo Mir Rivera. CIBERNED groups: G105. Type: CC.AA. Funding agency: Junta de Andalucía. Funding: Duraton: Code: PROPAG-AGEING Title: The continuum between healthy ageing and idiopathic Parkinson Disease within a propagation perspective of inflammation and damage: the search for new diagnostic, prognostic and therapeutic targets. Principal Investigator: Pablo Mir Rivera. CIBERNED groups: G105. Type: European. Funding agency: Comisión Europea. Funding: ,5. Duration: Code: RD12/0019/0008. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas (coordinador de red: José María Moraleda). CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RD16/0011/0017. Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas. CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: Axontherapix (Prorroga). Title: Axontherapix. Principal Investigator: José López Barneo. CIBERNED groups: G105. Other CIBER s collaboration: Yes (Redes de Terapia Celular). Type: National. Funding agency: Axontherapix. Funding: ,5. Duration: Code: Title: EMTICS (European Multicentre Tics in Children Studies). Principal Investigator: Pablo Mir Rivera. CIBERNED groups: G105. Type: European. Funding agency: Comisión Europea. Funding: ,6. Duration: Code: SAF P. Title: Fisiopatología de celulas madre derivadas de la cresta neural. Principal Investigator: Ricardo Pardal. CIBERNED groups: G105. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: Fundación Marcelino Botín. Title: Sensibilidad al oxígeno y neurodegeneracion. Principal Investigator: José López Barneo. CIBERNED groups: G105. Type: National. Funding agency: Fundación Marcelino Botin. Funding: Duration: Code: SAF2012/ Title: Sensibilidad al oxígeno y neurodegeneracion. Principal Investigator: José López Barneo. CIBERNED groups: G105. Type: National. Funding agency: Ministerio de Economía, Industrtia y Competividad. Funding: Duration: CIBERNED Annual Report / 166

167 Program 2 Group: José López Barneo Code: EMTICS 2011 (EUFP ). Title: European Multicentre Tics in Children Studies. Principal Investigator: Pablo Mir Rivera. CIBERNED groups: G105. Type: European. Funding agency: Comisión Europea. Funding: ,3. Duration: 2016 Code: PAI 2011(CTS516). Title: Fisiología Celular y Biofísica. Principal Investigator: José López Barneo. CIBERNED groups: G105. Type: CC.AA. Funding agency: Junta de Andalucía. Funding: 7.196,16. Duration: 2016 Code: ERC Title: Molecular mechanisms of acute oxygen sensing (OXYGENSENSING). Principal Investigator: José López Barneo. CIBERNED groups: G105. Type: European. Funding agency: Comisión Europea. Funding: Duration: Code: Pulmotension (II). Title: Pulmonary Hipertension: Functional Genomics and Therapy of Lung Vascular Remodelling. Principal Investigator: José López Barneo. CIBERNED groups: G105. Type: European. Funding agency: Comisión Europea. Funding: ,63. Duration: CIBERNED Annual Report / 167

168 GROUP Adolfo López de Munain 609 Principal Investigator Adolfo López de Munain Neurologist Research team Ana Aiastui Pujana PhD Garazi Aldanondo Aristizabal Miren Karmele Arnaiz Pérez Administrative staff Myriam Barandiarán Amillano Neurologist Alberto Bergareche Yarza M. Pilar Camaño González Leire Casas Fraile Rosalía Dacosta Aguayo PhD Manuel Delgado Alvarado Roberto Fernández Torrón Belén Gago Calderón PhD Federico García Bragado PhD Francisco Javier Gil Bea PhD María Goicoechea Bianchi PhD Ana Gorostidi Pagola PhD Haritz Jiménez Urbieta Biodonostia-Instituto de Investigación Sanitaria Área de Neurociencias Pº Dr. Beguiristain s/n - Planta 1ª San Sebastián (Spain) Guipúzcoa-España Tel.: Fax: mirenkarmele.arnaizperez@osakidetza.eus Jaione Lasa Elgarresta Haizpea Lasa Fernández José Félix Martí Masso PhD Pablo Martínez-Lage Álvarez Bachelor degree Alba Mateos Mateos Ayerdi Leyre Merino Galán Fermín Moreno Izco PhD Neia Naldaiz Gastezi Irene Navalpotro Gómez Nahikari Pastoriza Polo Technician Juanjo Poza Aldea PhD Ana Quiroga Valera PhD Tatiana Rodríguez Chinchilla María Cruz Rodríguez Oroz PhD Javier Ruiz Martínez PhD Amets Sáenz Peña PhD Andone Sistiaga Berrondo PhD Ivan Toral Ojeda PhD Ainara Vallejo Illarramendi PhD Mónica Zufiría García Miren Zulaica Ijurco PhD 2016 CIBERNED Annual Report / 168

169 Summary Main research lines: 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 multicenter clinical trial on ICD coordinated by this group is also under development, as well as a unicenter one. 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 Baque country) and of asymptomatic carriers of this mutation aiming to know pre-motor biomarkers of PD including clinical, molecular, 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 post-mortem 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 disease The group is composed by neurologists, neuropsychologists and biologists who develop a comprehensive approach to neuromuscular pathology from a clinical and basic aspect (including ELA and hereditary-based muscular diseases), as well as on the role of the muscle in the metabolic control and its influence on the development of central phenomena associated with aging (brain proteinopathies and aging). Main research lines: Myotonic dystrophy: a paper has been published about the incidence of cancer in the disease and the group continues the study of common metabolic 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. The study has resulted in a patent for some compounds called generically AHULKEN, valuing the creation of a start-up. Facioscapulohumeral dystrophy: the group continues collaborating with other groups in the clinical characterization of cases with scapuloperoneal syndrome and methylation patterns. Limb Girdle Dystrophy type 2 A: study of the influence of calpain 3 in proliferation, differentiation and replacement of muscle satellite cells, as well as the relationship with the extracellular matrix proteins. The group have published some collaborative and individual papers. Amyotrophic Lateral Sclerosis: the group has begun its journey in this field with the creation of an intraciber network that has obtained a project published a review. Keywords Parkinson Disease, Dyskinesias, impulse control disorder, dementia, cognitive impairment, LRRK2, neuroinflammation, muscular dystrophies, frontotemporal dementia, calpain, progranulin, biomarkers, neurogenetics, ALS 2016 CIBERNED Annual Report / 169

170 Publications Brockmann K, Schulte C, Schneiderhan-Marra N, Apel A, Pont-Sunyer C, Vilas D et al. Inflammatory profile discriminates clinical subtypes in LRRK2-associated Parkinson s disease. European journal of neurology. 2017;24(2). Epub Hamel W., Koppen J.A., Alesch F., Antonini A., Barcia J.A., Bergman H. et al. Targeting of the Subthalamic Nucleus for Deep Brain Stimulation: A Survey Among Parkinson Disease Specialists. World Neurosurgery. 2017;99: Epub Skrobot OA, O Brien J, Black S, Chen C, DeCarli C, Erkinjuntti T et al. The Vascular Impairment of Cognition Classification Consensus Study.Alzheimer s & dementia : the journal of the Alzheimer s Association Freyermuth F., Rau F., Kokunai Y., Linke T., Sellier C., Nakamori M. et al. Splicing misregulation of SCN5A contributes to cardiac-conduction delay and heart arrhythmia in myotonic dystrophy. Nature Communications. 2016;7. Fernandez-Torron R., Garcia-Puga M., Emparanza J.-I., Maneiro M., Cobo A.-M., Poza J.-J. et al. Cancer risk in DM1 is sex-related and linked to mirna-200/141 downregulation. Neurology. 2016;87(12): Fraser K.B., Moehle M.S., Alcalay R.N., West A.B., E. Tolosa, J.F. Martí Massó. Urinary LRRK2 phosphorylation predicts parkinsonian phenotypes in G2019S LRRK2 carriers. Neurology. 2016;86(11): Naldaiz-Gastesi N., Goicoechea M., Alonso-Martin S., Aiastui A., Lopez-Mayorga M., Garcia- Belda P. et al. Identification and Characterization of the Dermal Panniculus Carnosus Muscle Stem Cells. Stem Cell Reports. 2016;7(3): Alquezar C, Salado IG, de la Encarnación A, Pérez DI, Moreno F, Gil C et al. Targeting TDP- 43 phosphorylation by Casein Kinase-1δ inhibitors: a novel strategy for the treatment of frontotemporal dementia.molecular neurodegeneration. 2016;11(1). Delgado-Alvarado M., Gago B., Navalpotro-Gomez I., Jimenez-Urbieta H., Rodriguez-Oroz M.C. Biomarkers for dementia and mild cognitive impairment in Parkinson s disease. Movement Disorders. 2016;31(6): Marras C., Alcalay R.N., Caspell-Garcia C., Coffey C., Chan P., Duda J.E. et al. Motor and nonmotor heterogeneity of LRRK2-related and idiopathic Parkinson s disease. Movement Disorders Toral-Ojeda I., Aldanondo G., Lasa-Elgarresta J., Lasa-Fernandez H., Fernandez-Torron R., Lopez de Munain A. et al. Calpain 3 deficiency affects SERCA expression and function in the skeletal muscle. Expert Reviews in Molecular Medicine Garcia-Esparcia P, Koneti A, Rodríguez-Oroz MC, Lago B, Del Rio JA, Ferrer I. Mitochondrial activity in the frontal cortex area 8 and angular gyrus in Parkinson s disease and Parkinson s disease with dementia.brain pathology (Zurich, Switzerland) Muñoz-Culla M, Irizar H, Sáenz-Cuesta M, Castillo-Triviño T, Osorio-Querejeta I, Sepúlveda L et al. SncRNA (microrna &snorna) opposite expression pattern found in multiple sclerosis relapse and remission is sex dependent.scientific reports. 2016;6. Cervera-Carles L., Alcolea D., Estanga A., Ecay-Torres M., Izagirre A., Clerigue M. et al. Cerebrospinal fluid mitochondrial DNA in the Alzheimer s disease continuum. Neurobiology of Aging Tijms B.M., Kate M.T., Wink A.M., Visser P.J., Ecay M., Clerigue M. et al. Gray matter network disruptions and amyloid beta in cognitively normal adults. Neurobiology of Aging. 2016;37: ten Kate M., Sanz-Arigita E.J., Tijms B.M., Wink A.M., Clerigue M., Garcia-Sebastian M. et al. Impact of APOE-ɛ4 and family history of dementia on gray matter atrophy in cognitively healthy middle-aged adults. Neurobiology of Aging. 2016;38: Lopez de Maturana R., Lang V., Zubiarrain A., Sousa A., Vazquez N., Gorostidi A. et al. Mutations in LRRK2 impair NF-ΚB pathway in ipsc-derived neurons. Journal of Neuroinflammation. 2016;13(1). Brockmann K., Apel A., Schulte C., Schneiderhan-Marra N., Pont-Sunyer C., Vilas D. et al. Inflammatory profile in LRRK2-associated prodromal and clinical PD. Journal of Neuroinflammation. 2016;13(1) CIBERNED Annual Report / 170

171 Program 2 Group: Adolfo López de Munain Rodriguez-Perdigon M., Tordera R.M., Gil-Bea F.J., Gerenu G., Ramirez M.J., Solas M. Downregulation of glutamatergic terminals (VGLUT1) driven by Aβ in Alzheimer s disease. Hippocampus. 2016;26(10): Rodríguez-Arribas M, Pedro JM, Gómez-Sánchez R, Yakhine-Diop SM, Martínez-Chacón G, Uribe-Carretero E et al. Pompe Disease and Autophagy: Partners in Crime, or Cause and Consequence?. Current medicinal chemistry. 2016;23(21). Garcia-Garcia D, Guridi J, Toledo JB, Alegre M, Obeso JA, Rodríguez-Oroz MC. Stimulation sites in the subthalamic nucleus and clinical improvement in Parkinson s disease: a new approach for active contact localization.journal of neurosurgery Riancho J., Delgado-Alvarado M., Fernandez-Torre J.L., Sanchez-Juan P., Polo J.M. Subacute progressive aphasia: a rare presentation of Creutzfeldt Jakob disease. Journal of Neurology. 2016; 1-3. Evangelista T., Wood L., Fernandez-Torron R., Williams M., Smith D., Lunt P. et al. Design, setup and utility of the UK facioscapulohumeral muscular dystrophy patient registry. Journal of Neurology. 2016;263(7): Fernandez-Costa J.M., Llamusi B., Bargiela A., Zulaica M., Alvarez-Abril M.C., Perez-Alonso M. et al. Six serum mirnas fail to validate as myotonic dystrophy type 1 biomarkers. PLoS ONE. 2016;11(2). Abete I., Zulet M.A., Goyenechea E., Blazquez V., de Arce Borda A.M., Lopez de Munain A. et al. Association of lifestyle, inflammatory factors, and dietary patterns with the risk of suffering a stroke: A case control study. Nutritional Neuroscience. 2016; 1-9. Gorostidi A., Marti-Masso J.F., Bergareche A., Rodriguez-Oroz M.C., de Munain A.L., Ruiz- Martinez J. Genetic Mutation Analysis of Parkinson s Disease Patients Using Multigene Next- Generation Sequencing Panels. Molecular Diagnosis and Therapy. 2016; Navalpotro-Gomez I., Vivanco-Hidalgo R.M., Cuadrado-Godia E., Medrano-Martorell S., Alameda- Quitllet F., Villalba-Martinez G. et al. Focal status epilepticus as a manifestation of idiopathic hypertrophic cranial pachymeningitis. Journal of the Neurological Sciences. 2016;367: Marcos-Gragera R., Galceran J., Martos C., de Munain A.L., Vicente-Raneda M., Navarro C. et al. Incidence and survival time trends for Spanish children and adolescents with leukaemia from 1983 to Clinical and Translational Oncology. 2016; Blanco Martin E., Ugarriza Serrano I., Elcoroaristizabal Martin X., Galdos Alcelay L., Molano Salazar A., Bereincua Gandarias R. et al. Dysexecutive syndrome in amnesic mild cognitive impairment: A multicenter study. BMC Neurology. 2016;16(1). Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Grandal Leiros B., Perez Mendez L.I., Zelaya Huerta M.V., Moreno Eguinoa L., Garcia-Bragado F., Tunon Alvarez T. et al. Prevalence and concordance between the clinical and the postmortem diagnosis of dementia in a psychogeriatric clinic. Neurologia Andrés N, Poza JJ, Martí Massó JF. Familial amyloidosis with polyneuropathy type 1 caused by transthyretin mutation Val50Met (Val30Met): 4 cases in a non-endemic area.neurologia (Barcelona, Spain) Garcia-Ribas G., Arbizu J., Carrio I., Garrastachu P., Martinez-Lage P. PET biomarkers: Use of imaging techniques in Alzheimer disease and neurodegeneration clinical diagnosis. Neurologia Santurtun A., Villar A., Delgado-Alvarado M., Riancho J. Trends in motor neuron disease: association with latitude and air lead levels in Spain. Neurological Sciences. 2016; 1-5. Barandika O., Irigoyen C., Anasagasti A., Egiguren G., Ezquerra-Inchausti M., Lopez De Munain A. et al. A Cost-Effective Mutation Screening Strategy for Inherited Retinal Dystrophies. Ophthalmic Research. 2016;56(3): Santurtún A, Delgado-Alvarado M, Villar A, Riancho J. [Geographical distribution of mortality by Parkinson s disease and its association with air lead levels in Spain].Medicina clinica CIBERNED Annual Report / 171

172 Poza-Aldea J.J. A proposal for a model to replace carbamazepine or oxcarbazepine by eslicarbazepine acetate in clinical practice. Revista de Neurologia. 2016;63(5): Tainta M., de la Riva P., Gonzalez F., Marti-Masso J.F., Goenaga M.A. Rhomboencephalitis and endocarditis caused by listeria monocytogenes: An unreported association. Revista de Neurologia. 2016;63(1): Castiella A, Zapata E, De Juan MD, Zubiaurre L, Iribarren A, Otazua P et al. Analysis of HFE mutations and non-hfe gene mutations (TFR2 and SLC40A1) in patients with phenotypic hemochromatosis from the Basque Country.International journal of laboratory hematology. 2016;38(1). Munoz-Canoves P., Carvajal J.J., de Munain A.L., Izeta A. Editorial: Role of stem cells in skeletal muscle development, regeneration, repair, aging, and disease. Frontiers in Aging Neuroscience. 2016;8(APR). Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. De la Riva P, Andres-Marín N, Gonzalo-Yubero N, Tainta-Cuezva M, Caminos N, Urtasun- Ocariz MÁ et al. Headache and other complications following intrathecal chemotherapy administration.cephalalgia : an international journal of headache Sáenz A, López de Munain A. Dominant LGMD2A: alternative diagnosis or hidden digenism?. Brain : a journal of neurology Richard I, Hogrel JY, Stockholm D, Payan CA, Fougerousse F, Calpainopathy Study Group. et al. Natural history of LGMD2A for delineating outcome measures in clinical trials.annals of clinical and translational neurology. 2016;3(4). Research projects Code: PI2014/06. Title: Inicio y progresión de la enfermedad de Parkinson: papel de la activación glial. Principal Investigator: Mª Cruz Rodríguez Oroz. CIBERNED groups: G609. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: AGL REDT. Title: Red nacional de priones. Principal Investigator: José Antonio del Río. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G609, G207. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: PI2016/04. Title: The ALS CIBERNED challenge: accelerating new drug discovery. Principal Investigator: Adolfo López De Munain Arregui. CIBERNED s collaboration: Yes. CIBERNED groups: G609, G303, G503, G408. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: DTS15/ Title: Evaluación del impacto de la imagen PET de amiloide en el diagnóstico de los pacientes con deterioro cognitivo evaluados por sospecha de Alzheimer. Principal Investigator: Dr. Javier Arbizu. CIBERNED s collaboration: Yes. CIBERNED groups: G504, G502, G609. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: 2016 Code: RTC Title: Desarrollo de agentes terapéuticos basados en ácidos nucléicos para el tratamiento de enfermedades neuromotoras y neuromusculares. Principal Investigator: Rubén Lépez Vales. CIBERNED s collaboration: Yes. CIBERNED groups: G607, G609. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 172

173 Program 2 Group: Adolfo López de Munain Code: PI14/ Title: Estudio de la capacidad regenerativa de los progenitores musculares derivados del ipss del pacientes con distrofia de cinturas tipo 2ª. Estudio in vitro y en un modelo murino de dano tisular. Principal Investigator: Adolfo López De Munain. CIBERNED groups: G609. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: UE/2014/CAPN3_CA. Title: Targeting calcium handling proteins in LGMD2A muscular dystrophy. Principal Investigator: Ainara Vallejo. CIBERNED groups: G609. Type: International. Funding agency: Association Française Contre Les Myopathies. Funding: Duration: Code: FMM Title: Testado de nuevas sustancias con potencial terapéutico in vitro e in vivo para el tratamiento de la distrofia muscular de Duchenne. Principal Investigator: Ainara Vallejo. CIBERNED groups: G609. Type: International. Funding agency: Asociacion Duchenne Parent Project España. Funding: Duration: Code: Title: Implicación de la calpaina 3 en la senescencia prematura de las células satelite musculares: estudio del ciclo celular en progenitores musculares derivados de IPSs de pacientes con LGMD2A. Principal Investigator: Adolfo López de Munain. CIBERNED groups: G609. Type: CC.AA. Funding agency: Gobierno Vasco. Funding: Duration: Code: Title: La esclerosis lateral amiotrófica como enfermedad metabólica: estudio de mecanismos patogénicos y aproximación terapéutica. Principal Investigator: Francisco Javier Gil Bea. CIBERNED groups: G609. Type: CC.AA. Funding agency: Gobierno Vasco. Funding: Duration: PhD dissertations Author: Fermín Moreno Izco. Title: Demencia frontotemporal por mutaciones en el gen de la progranulina. Aspectos clínicos y moleculares. Date: 28 Octobre Supervisor: Adolfo López de Munain Arregui. Author: Myriam Barandiaran Amillano. Title: Estudio longitudinal y caracterización cognitiva de portadores de la mutación C.709-1G>A en el gen de la Progranulina. Date: 28 Octobre Supervisor: Adolfo López de Munain Arregui. Author: Ivan Toral Ojeda. Title: Study of calcium-handling proteins in Limb-girdle muscular dystrophy type 2A. Date: 22 July Supervisor: Ainara Vallejo Illarramendi. Author: Sara Bahillo Santamaría. Title: Papel del ácido úrico en la enfermedad de Parkinson asociada o no a mutaciones en el gen LRRK2. Date: 15 June Supervisor: Javier Ruiz Martínez. Author: Carmen Gasca Salas. Title: Estudio longitudinal del deterioro cognivo en la enfermedad de Parkinson. Date: 8 February Supervisor: María Cruz Rodríguez Oroz CIBERNED Annual Report / 173

174 GROUP José Javier Lucas Lozano 306 Principal Investigator José Javier Lucas Lozano Research team María Santos Galindo Researcher Jorge Rubén Cabrera Heredia Postdoctoral fellow Ainara Elorza Peregrina Postdoctoral fellow Marta Fernández Nogales Alberto Parras Rodríguez Ivó Hernández Hernández Sara Picó del Pino Student Miriam Lucas Santamaría Researcher Centro de Biología Molecular Severo Ochoa CSIC/UAM c/nicolás Cabrera, 1, Madrid (Spain) Tel.: / (Lab) Fax: CIBERNED Annual Report / 174

175 Summary Huntington s disease (HD) is the most prevalent genetic neurodegenerative disease, caused by an expansion of the trinucleotide CAG located in 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. This way, we discovered an isoform imbalance in tau, a protein related with Alzheimer s disease and other dementias, and a new histopathological hallmark (the Tau Nuclear Rods or TNRs), consisting on the presence of tau in nuclear indentations, in HD. Furthermore, the splicing factor SRSF6, involved in tau splicing and capable to binding CAGrepeats, is altered in HD and sequestered in the mutant huntingtin inclusion bodies (Nat Med. 2014, 20(8):881-5). In order to characterize if tau alteration is enough for TNR appearance, we have tested a mouse model of FTDP17, a disease caused by altered tau. The TNR appearance in this model let us conclude that TNR detection could be useful as marker of tau imbalance (Brain Pathol. doi: /bpa.12407). We have also described decreased GSK3b, a kinase able to phosphorylate tau, in HD. Disease phenotype is reversible with GSK3b overexpression in an HD model (HMG 24(17): ). As well, we have studied levels and isoforms of MAP2, another microtubuleassociated protein which is related to tau. Its levels drop, its isoforms are imbalanced and its subcellular localization is aberrant in HD. In cell models, this splicing can be performed by SRSF6 (Brain Pathol. 27(2): ). These splicing alterations in HD have opened a new research line in which we are currently working in our lab. Keywords Huntington s disease, transgenic mice, GSK-3, Tau, MAP2, SRSF6, splicing Publications Cabrera JR, Lucas JJ. MAP2 Splicing is Altered in Huntington s Disease.Brain pathology (Zurich, Switzerland) Fernandez-Nogales M., Santos-Galindo M., Merchan-Rubira J., Hoozemans J.J.M., Rabano A., Ferrer I. et al. Tau-positive nuclear indentations in P301S tauopathy mice. Brain Pathology Fernandez-Nogales M., Santos-Galindo M., Hernandez I.H., Cabrera J.R., Lucas J.J. Faulty splicing and cytoskeleton abnormalities in Huntington s disease. Brain Pathology. 2016;26(6): Sorolla MA, Rodríguez-Colman MJ, Vall-Llaura N, Vived C, Fernández-Nogales M, Lucas JJ et al. Impaired PLP-dependent metabolism in brain samples from Huntington disease patients and transgenic R6/1 mice.metabolic brain disease. 2016;31(3). de Diego-García L, Ramírez-Escudero M, Sebastián-Serrano Á, Diaz-Hernández JI, Pintor J, Lucas JJ et al. Regulation of proteasome activity by P2Y₂ receptor underlies the neuroprotective effects of extracellular nucleotides.biochimica et biophysica acta. 2016;1863(1). Research projects Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz Cánoves. CIBERNED s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 175

176 GROUP Rosario Moratalla Villalba 204 Principal Investigator Rosario Moratalla Villalba Research team Noelia Granado Martínez Postdoctoral fellow Luz María Suárez PhD Patricia García Sanz PhD José Rubén García Montes Postdoctoral fellow Irene Ruiz De Diego Postdoctoral fellow Oscar Solís Castrejón Lorena Orgaz Gordillo Guillermo Bueno Gil Samuel Alberquilla Researcher staff Emilia Rubio Rubio Lab technician Beatriz Pro Lab technician Instituto Cajal (CSIC) Avenida Dr. Arce, Madrid (Spain) Tel.: / Fax: moratalla@cajal.csic.es 2016 CIBERNED Annual Report / 176

177 Summary During this year 2016, we have demonstrated the changes in synaptic transmission in striatal projection neurons (MSNs) in dyskinetic mice correlate with changes in spine morphology, rather than with changes in spine density (Figure 1). In particular, we show that in direct neurons (D1- MSNs) the synaptic transmission is maintained, despite the fact that L-DOPA does not alter the spine-pruning caused by dopaminergic loss. After chronic L-DOPA treatment, the remaining spines exhibit increased volume, length, and head diameter, with higher PSD-content, which allows a greater input of information into the dendrite maintaining the synaptic transmission in normal values (Suarez et al 2016). In contrast, in D2-MSNs, chronic L-DOPA treatment restores spine density, reduced by dopamine depletion. However, the newly born spines are not fully functional due to lower PSD content and thinner necks which prevent synaptic transmission from fully recovering. Then, abnormal spine morphology correlates with dyskinetic behaviors. Also, we established that L-DOPA restores back to normal the increased amino acid levels in the striatum of 6-OHDAlesioned mice, in line with the synaptic plasticity. Moreover, L-DOPA produces a strong increase in GABA and tyrosine levels that correlate with the hyperactivation of D1R-containing striatal neurons and with the appearance of striatal THpositive neurons, respectively (Solis et al 2016). This year, we also demonstrate that the C-terminal domain of the heavy chain of tetanus toxin (Hc-TeTx) is a nontoxic peptide can provide significant protection against acute methamphetamine, a derivated amphetamine drug that induced dopaminergic damage, induced neurotoxicity (Figure 2) and motor impairment, suggesting its therapeutic potential in methamphetamine abusers (Mendieta et al 2016). Keywords Parkinson s disease, tyrosine, spines, GABA, methamphetamine, neurotoxicity Publications Suarez LM, Solis O, Aguado C, Lujan R, Moratalla R. L-DOPA Oppositely Regulates Synaptic Strength and Spine Morphology in D1 and D2 Striatal Projection Neurons in Dyskinesia.Cerebral cortex (New York, N.Y. : 1991) Solis O., Garcia-Sanz P., Herranz A.S., Asensio M.-J., Moratalla R. L-DOPA Reverses the Increased Free Amino Acids Tissue Levels Induced by Dopamine Depletion and Rises GABA and Tyrosine in the Striatum. Neurotoxicity Research. 2016; 1-9. Suarez L.M., Munoz M.-D., Gonzalez J.C., Bustamante J., Del Rio R.M., Solis J.M. The taurine transporter substrate guanidinoethyl sulfonate mimics the action of taurine on long-term synaptic potentiation. Amino Acids. 2016; Mendieta L., Granado N., Aguilera J., Tizabi Y., Moratalla R. Fragment C domain of tetanus toxin mitigates methamphetamine neurotoxicity and its motor consequences in mice. International Journal of Neuropsychopharmacology. 2016;19(8):1-11. Suarez L.M., Munoz M.-D., Martin Del Rio R., Solis J.M. Taurine content in different brain structures during ageing: Effect on hippocampal synaptic plasticity. Amino Acids. 2016;48(5): Research projects 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. CIBERNED groups: G204. Type: International. Funding agency: Secretaria de Ciencia, Tecnologia e Innovacion del Distrito Federal, Mexico. Funding: Duration: CIBERNED Annual Report / 177

178 Code: PI2015-2/02. Title: Potencial patológico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal Investigator: Joan X. Comella Carnice. CIBERNED s collaboration: Yes. CIBERNED groups: G413, G415, G204, G108, G411. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: Cajal Blue Brain Project. Title: Cajal Blue Brain Project. International Blue Brain Proyect. Principal Investigation: Javier de Felipe. CIBERNED s collaboration: Yes. CIBERNED groups: G204, G403. Type: International. Funding agency: Ecole Polytechnique Federale de Lausanne (Suiza), IBM, Universidad Politécnica de Madrid, Consejo Superior de Investigaciones Cientificas, España. Funding: N.D. Duration: Code: SAF R. Title: Mecanismos moleculares responsables de la plasticidad sináptica y remodelación estructural de las neuronas estriatales de proyección en ratones disquinéticos. Principal Investigator: Rosario Moratalla. CIBERNED groups: G204. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: PCIN Title: Development of a new in vivo radiotracer for alpha-synuclein. Principal Investigator: Mireille Dumoilin. CIBERNED groups: G204. Type: European. Funding agency: Comisión Europea. Funding: Duration: CIBERNED Annual Report / 178

179 GROUP Pura Muñoz-Cánoves 604 Principal Investigator Pura Muñoz Cánoves ICREA, research professor Research team Antonio L. Serrano Sánchez Senior researcher Eusebio Perdiguero Santamaría Senior researcher, CIBERNED Mercè Jardí Ripoll Technician Vera Lukesova Technician Laura Ortet Postdoctoral fellow Marina Raya Labmanager Jessica Segales Dalmau Postdoctoral fellow Begoña Ampudia Carrasco Animal facility technician Yacine Kharraz Postdoctoral fellow Vanessa Ruíz-Bonilla Postdoctoral fellow, CIBERNED Laura García Prat Antonio Martínez Pre-doctoral fellow Victoria Moiseeva Pre-doctoral fellow Cell Biology Group ICREA and Pompeu Fabra University (UPF) Department of Experimental and Life Sciences (DCEXS) Tel.: Fax: CIBERNED Annual Report / 179

180 Summary Muscle stem cell regenerative decline with aging. Muscle stem cells lose autophagic activity as they age and this promotes senescence entry: We have shown that basal autophagy is indispensable to maintain the normal stem-cell quiescent state in mice. Failure of autophagy in physiologically aged satellite cells causes senescence entry and increased mitochondrial dysfunction, resulting in numerical and functional satellite cell decline. Autophagy reestablishment reverses senescence and restores regenerative functions in geriatric satellite cells. These findings uncover autophagy as a decisive stem cell-fate regulator and have implications for fostering muscle regeneration in sarcopenia. García-Prat L, Martínez-Vicente M, Perdiguero E, Ortet L, Garcia-Ubreva J, Rebollo E, Gutarra S, Ballestar E, Serrano AL, Sandri M, Muñoz-Cánoves P. Autophagy maintains stemness by preventing senescence. Nature 529: 37-42, full commentaries/articles in: Autophagy 2016 doi: / ; Bioessays 2016 doi: /bies ; Rejuvenation Res 2016 doi: / rej ; Oral Dis doi: / odi Nature Revs Mol Biol. nrm html; - 1 invited Article Summary in Nature News & Views doi: / nature Faculty of 1000 Prime (F1000Prime) f1000.com/prime/ García-Prat L, Martínez-Vicente M, Muñoz- Cánoves P. Autophagy: a decisive process for stemness. Oncotarget 7, García-Prat L, Muñoz-Cánoves P*, Martínez- Vicente M*. Dysfunctional autophagy is a driver of muscle stem cell decline with aging. Autophagy 12:612-3, 2016 (*cocorresponding authors). Additional studies on stem cells and striated muscle: We have shown a p38amapk-dependent epigenetic regulation of muscle stem cells during myogenesis. Segalés J, Islam AB, Sousa-Victor P, Dilworth FJ, Perdiguero E, Muñoz-Cánoves P. Chromatin-wide and transcriptome profiling integration uncovers p38α MAPK as a global regulator of skeletal muscle differentiation. Skelet Muscle 15;6:9, Collaborations on the mechanisms regulating skeletal and cardiac muscle functions: Gómez del Arco at al. Cell Metabol 23:881-92, Keywords Skeletal muscle, regeneration, muscle stem cells, sarcopenia, aging, inflammation, fibrosis Publications Sousa-Victor P., Munoz-Canoves P. Regenerative decline of stem cells in sarcopenia. Molecular Aspects of Medicine Sebastian D., Sorianello E., Segales J., Irazoki A., Ruiz-Bonilla V., Sala D. et al. Mfn2 deficiency links age-related sarcopenia and impaired autophagy to activation of an adaptive mitophagy pathway. EMBO Journal García-Prat L, Muñoz-Cánoves P, Martinez-Vicente M. Dysfunctional autophagy is a driver of muscle stem cell functional decline with aging.autophagy Gomez-Del Arco P., Perdiguero E., Yunes-Leites P.S., Acin-Perez R., Zeini M., Garcia-Gomez A. et al. The Chromatin Remodeling Complex Chd4/NuRD Controls Striated Muscle Identity and Metabolic Homeostasis. Cell Metabolism. 2016;23(5): Serrano AL, Muñoz-Cánoves P. Fibrosis development in early-onset muscular dystrophies: Mechanisms and translational implications.seminars in cell & developmental biology García-Prat L, Muñoz-Cánoves P. Aging, metabolism and stem cells: Spotlight on muscle stem cells.molecular and cellular endocrinology CIBERNED Annual Report / 180

181 Program 2 Group: Pura Muñoz-Cánoves Garcia-Prat L., Martinez-Vicente M., Munoz-Canoves P. Autophagy: A decisive process for stemness. Oncotarget. 2016;7(11): Brack A.S., Munoz-Canoves P. The ins and outs of muscle stem cell aging. Skeletal Muscle Segales J., Islam A.B.M.M.K., Kumar R., Liu Q.-C., Sousa-Victor P., Dilworth F.J. et al. Chromatinwide and transcriptome profiling integration uncovers p38α MAPK as a global regulator of skeletal muscle differentiation. Skeletal Muscle. 2016;6(1). Garcia-Prat L., Martinez-Vicente M., Munoz-Canoves P. Methods for mitochondria and mitophagy flux analyses in stem cells of resting and regenerating skeletal muscle. Methods in Molecular Biology. 2016;1460: Pessina P., Munoz-Canoves P. Fibrosis-Inducing strategies in regenerating dystrophic and normal skeletal muscle. Methods in Molecular Biology. 2016;1460: Segalés J, Perdiguero E, Muñoz-Cánoves P. Regulation of Muscle Stem Cell Functions: A Focus on the p38 MAPK Signaling Pathway.Frontiers in cell and developmental biology. 2016;4. Munoz-Canoves P., Carvajal J.J., de Munain A.L., Izeta A. Editorial: Role of stem cells in skeletal muscle development, regeneration, repair, aging, and disease. Frontiers in Aging Neuroscience. 2016;8. Medici D., Munoz-Canoves P., Yang P.-C., Brunelli S. Mesenchymal Transitions in Development and Disease. Stem Cells International. 2016;2016. Kharraz Y., Lefort A., Libert F., Mann C.J., Gueydan C., Kruys V. Genome-wide analysis of TIAR RNA ligands in mouse macrophages before and after LPS stimulation. Genomics Data. 2016;7: Research projects Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz-Cánoves. CIBERNED s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: AGAUR SGR 102. Title: Biología Celular. Principal Investigator: Pura Muñoz-Cánoves. CIBERNED groups: G604. Type: CC.AA. Funding agency: AGAUR-Generalitat de Catalunya. Funding: Duration: Code: ERARE13-fp-149. Title: Stimulating Intrinsic Repair for DMD (SIRD). Principal Investigator: Michael Rudnicki. CIBERNED groups: G604. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: MDA Title: Understanding and reversing muscle stem cell regenerative decline in DMD. Principal Investigation: Pura Muñoz Cánoves. CIBERNED groups: G604. Type: International. Funding agency: Muscular Dystrophy Association (MDA)-USA. Funding: Duration: CIBERNED Annual Report / 181

182 Code: SAF R). Title: Understanding skeletal muscle regenerative decline with aging. Principal Investigation: Pura Muñoz- Cánoves. CIBERNED groups: G604. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: 2014 SGR 102. Title: Understanding the basis of muscle regeneration. Principal Investigator: Pura Muñoz-Cánoves. CIBERNED groups: G604. Type: CC.AA. Funding agency: AGAUR-Generalitat Catalunya. Funding: Duration: PhD dissertations Author: Joana Guerra Badell. Title: Mechanisms regulating skeletal muscle growth and atrophy. Date: 14 October Supervisor: Antonio L. Serrano Sánchez. Author: Diana Sobral Mesquita. Title: Regulation of Skeletal Muscle Regeneration and Growth. Date: 19 September Supervisor: Pura Muñoz Cánoves. Author: Pedro Maseres. Title: Regulation of myogenesis by p38 MAPK signaling. Date: 20 June Supervisor: Pura Muñoz Cánoves CIBERNED Annual Report / 182

183 GROUP José Ramón Naranjo Orovio 307 Principal Investigator José Ramón Naranjo Research team Britt Mellström CIBERNED researcher Alberto Rábano F.CIEN-CIBERNED researcher Alejandro López-Hurtado Máster-UAM, student Daniel Fernández Máster-UAM, student Rocio Naranjo TFG-UAM, student M. Paz González Assistant researcher Xosé Manuel Dopazo Assistant researcher CSIC. Centro Nacional de Biotecnología Molecular and Cellular Biology c/darwin, Madrid (Spain) Tel.: naranjo@cnb.csic.es 2016 CIBERNED Annual Report / 183

184 Summary We investigate the nuclear components of activity- and Ca2+-dependent transcriptional responses in neurons to: 1) 2) Understand the molecular determinants of downstream events that are responsible for early plastic changes in synaptic functionality during neurodegeneration. 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 and protein homeostasis are common features of some neurodegenerative pathologies, including Alzheimer s (AD), Down syndrome (DS) and Huntington s disease (HD), pathologies that are in the focus of our work. 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+ and protein homeostasis. Importantly, an early reduction in DREAM levels is found in mouse models for these pathologies (AD, DS and HD) and, genetic experiments show that this could be part of a neuroprotective mechanism operating in HD. The mechanism involves the protein-protein interaction between DREAM and ATF6, a key protein in the early activation of the unfolded protein response (UPR), as well as the regulation at the transcriptional level of several cytoskeletal proteins that are important for axonal transport and synaptic function. 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 Publications Garcia-Ayllon M.-S., Botella-Lopez A., Cuchillo-Ibanez I., Rabano A., Andreasen N., Blennow K. et al. HNK-1 Carrier Glycoproteins Are Decreased in the Alzheimer s Disease Brain. Molecular Neurobiology. 2016; Fernandez-Nogales M., Santos-Galindo M., Merchan-Rubira J., Hoozemans J.J.M., Rabano A., Ferrer I. et al. Tau-positive nuclear indentations in P301S tauopathy mice. Brain Pathology Pajares M., Jimenez-Moreno N., Garcia-Yague A.J., Escoll M., de Ceballos M.L., Van Leuven F. et al. Transcription factor NFE2L2/NRF2 is a regulator of macroautophagy genes. Autophagy. 2016; De Pedro-Cuesta J., Martinez-Martin P., Rabano A., Alcalde-Cabero E., Jose Garcia Lopez F., Almazan-Isla J. et al. Drivers: A biologically contextualized, cross-inferential view of the epidemiology of neurodegenerative disorders. Journal of Alzheimer s Disease. 2016;51(4): de Pedro-Cuesta J., Martinez-Martin P., Rabano A., Ruiz-Tovar M., Alcalde-Cabero E., Calero M. Etiologic framework for the study of neurodegenerative disorders as well as vascular and metabolic comorbidities on the grounds of shared epidemiologic and biologic features. Frontiers in Aging Neuroscience. 2016;8(JUN). Benedet T, Gonzalez P, Oliveros JC, Dopazo JM, Ghimire K, Palczewska M et al. Transcriptional repressor DREAM regulates trigeminal noxious perception.journal of neurochemistry Sanchez-Mut J.V., Heyn H., Vidal E., Moran S., Sayols S., Delgado-Morales R. et al. Human DNA methylomes of neurodegenerative diseases show common epigenomic patterns. Translational Psychiatry. 2016;6(1) CIBERNED Annual Report / 184

185 Program 2 Group: José Ramón Naranjo Orovio Rabano A., Cuadros R., Merino-Serrais P., Rodal I., Benavides-Piccione R., Gomez E. et al. Protocols for monitoring the development of tau pathology in Alzheimer s disease. Methods in Molecular Biology. 2016;1303: Lastres-Becker I., Garcia-Yague A.J., Scannevin R.H., Casarejos M.J., Kugler S., Rabano A. et al. Repurposing the NRF2 Activator Dimethyl Fumarate as Therapy Against Synucleinopathy in Parkinson s Disease. Antioxidants and Redox Signaling. 2016;25(2): Naranjo J.R., Zhang H., Villar D., Gonzalez P., Dopazo X.M., Moron-Oset J. et al. Activating transcription factor 6 derepression mediates neuroprotection in Huntington disease. Journal of Clinical Investigation. 2016;126(2): Mellstrom B., Kastanauskaite A., Knafo S., Gonzalez P., Dopazo X.M., Ruiz-Nuno A. et al. Specific cytoarchitectureal changes in hippocampal subareas in dadream mice. Molecular Brain. 2016;9(1). 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 groups: G111, G401, G104, G307, G409. Other CIBER s collaboration: Yes (CIBERER). Type: CC.AA. Funding agency: Comunidad de Madrid. Funding: Duration: Code: PI2016/05. Title: Dream inhibitors and Alzheimer s disease. Principal Investigator: José Ramón Naranjo Orovio. CIBERNED s collaboration: Yes. CIBERNED groups: G307, G106, G403. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: SAF R. Title: Mecanismos de acción y propiedades terapéuticas de los ligandos de DREAM. Principal Investigator: José Ramón Naranjo Orovio. CIBERNED groups: G307. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: 2016 Code: RTC Title: Validación de DREAM como nueva diana terapéutica para el tratamiento de la enfermedad de Alzheimer y desarrollo experimental de moléculas candidatas a ser nuevos productos terapéuticos. Principal Investigator: Antonio Gómez (JANSSEN-CILAG). CIBERNED groups: G307. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 185

186 GROUP Xavier Navarro Acebes 607 Principal Investigator Xavier Navarro Acebes Full professor Research team Esther Udina Bonet Associate professor, researcher Rubén López Vales Associate professor, researcher Caty Casas Louzao Associate professor, researcher Assumpció Bosch Merino Associate professor, researcher Guillermo García Alías Ramón y Cajal researcher Jordi Cuadras Mas Emeritous professor, researcher Jordi Bruna Escuer Assistant professor Roser Velasco Fargas Assistant professor Stefano Cobianchi Postdoctoral fellow Jaume del Valle Macià Postdoctoral fellow Joaquim Hernández Martín Postdoctoral fellow Mireia Herrando Grabulosa Postdoctoral fellow Clara Penas Pérez Postdoctoral fellow Jesús Amo Aparicio Ariadna Arbat Plana Natalia De la Oliva Muñoz Isaac Francos Quijorna Nuria Gaja Capdevila Tatiana Leiva Rodríguez Víctor Manuel López Álvarez Clara López Serrano Anna Martínez Muriana María Puigdomenech Poch David Romeo Guitart Alba Sánchez Fernández Daniel Santos Rojas Jordi Badia Casahuja Researcher Miguel Ángel Rubio Pérez Researcher Jessica Jaramillo Rodríguez Technician Israel Blasco Ruano Technician, CIBERNED Dept. Cell Biology, Physiology and Immunology Institute of Neurosciences Universitat Autònoma de Barcelona Avenida Can Domènech, edificio M, campus UAB Bellaterra, Catalunya (Spain) Tel.: Fax: xavier.navarro@uab.cat 2016 CIBERNED Annual Report / 186

187 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, neural 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. Neuromodulation of neural plasticity for promoting functional restitution in spinal cord lesions. 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, neuroplasticity, neural interfaces Publications Guiho T, Andreu D, López-Alvarez VM, Cvancara P, Hiairrassary A, Granata G, et al. Advanced 56 Channels Stimulation System to Drive Intrafascicular Electrodes. In: Converging Clinical and Engineering Research on Neurorehabilitation II. Ibáñez J, González-Vargas J, Azorín J, Akay M, Pons J, editors. Biosystems & Biorobotics, Springer, 2017, vol 15, pp ISBN: In press Coll-Miro M., Francos-Quijorna I., Santos-Nogueira E., Torres-Espin A., Bufler P., Dinarello C.A. et al. Beneficial effects of IL-37 after spinal cord injury in mice. Proceedings of the National Academy of Sciences of the United States of America. 2016;113(5): Meyer C, Stenberg L, Gonzalez-Perez F, Wrobel S, Ronchi G, Udina E et al. Chitosan-film enhanced chitosan nerve guides for long-distance regeneration of peripheral nerves. Biomaterials. 2016;76. Sanchez-Osuna M., Martinez-Escardo L., Granados-Colomina C., Martinez-Soler F., Pascual- Guiral S., Iglesias-Guimarais V. et al. An intrinsic DFF40/CAD endonuclease deficiency impairs oligonucleosomal DNA hydrolysis during caspase-dependent cell death: A common trait in human glioblastoma cells. Neuro-Oncology. 2016;18(7): CIBERNED Annual Report / 187

188 Izquierdo C., Velasco R., Vidal N., Sanchez J.J., Argyriou A.A., Besora S. et al. Lymphomatosis cerebri: A rare form of primary central nervous system lymphoma. Analysis of 7 cases and systematic review of the literature. Neuro-Oncology. 2016;18(5): Francos-Quijorna I., Amo-Aparicio J., Martinez-Muriana A., Lopez-Vales R. IL-4 drives microglia and macrophages toward a phenotype conducive for tissue repair and functional recovery after spinal cord injury. GLIA Santos D, Wieringa P, Moroni L, Navarro X, Valle JD. PEOT/PBT Guides Enhance Nerve Regeneration in Long Gap Defects.Advanced healthcare materials Martinez-Muriana A., Mancuso R., Francos-Quijorna I., Olmos-Alonso A., Osta R., Perry V.H. et al. CSF1R blockade slows the progression of amyotrophic lateral sclerosis by reducing microgliosis and invasion of macrophages into peripheral nerves. Scientific Reports. 2016;6. Olivan S., Calvo A.C., Rando A., Herrando-Grabulosa M., Manzano R., Zaragoza P. et al. Neuroprotective effect of non-viral gene therapy treatment based on tetanus toxin C-fragment in a severe mouse model of spinal muscular atrophy. Frontiers in Molecular Neuroscience. 2016;9. Mancuso R, Martínez-Muriana A, Leiva T, Gregorio D, Ariza L, Morell M et al. Neuregulin-1 promotes functional improvement by enhancing collateral sprouting in SOD1(G93A) ALS mice and after partial muscle denervation.neurobiology of disease. 2016;95. Redondo-Castro E., Navarro X., Garcia-Alias G. Longitudinal evaluation of residual cortical and subcortical motor evoked potentials in spinal cord injured rats. Journal of Neurotrauma. 2016;33(10): Cobianchi S., Arbat-Plana A., Lopez-Alvarez V.M., Navarro X. Neuroprotective effects of exercise treatments after injury: The dual role of neurotrophic factors. Current Neuropharmacology. 2016;14. Santos D., Gonzalez-Perez F., Navarro X., Del Valle J. Dose-Dependent Differential Effect of Neurotrophic Factors on in Vitro and in Vivo Regeneration of Motor and Sensory Neurons. Neural Plasticity. 2016;2016. López-Serrano C, Torres-Espín A, Hernández J, Alvarez-Palomo AB, Requena J, Gasull X et al. Effects of the spinal cord injury environment on the differentiation capacity of human neural stem cells derived from induced pluripotent stem cells.cell transplantation Delgado-Martinez I., Badia J., Pascual-Font A., Rodriguez-Baeza A., Navarro X. Fascicular topography of the human median nerve for neuroprosthetic surgery. Frontiers in Neuroscience. 2016;10. Herrando-Grabulosa M, Mulet R, Pujol A, Mas JM, Navarro X, Aloy P et al. Novel Neuroprotective Multicomponent Therapy for Amyotrophic Lateral Sclerosis Designed by Networked Systems. PloS one. ;11(1). Gonzalez-Fernandez C., Mancuso R., Del Valle J., Navarro X., Rodriguez F.J. Wnt signaling alteration in the spinal cord of amyotrophic lateral sclerosis transgenic mice: Special focus on Frizzled-5 cellular expression pattern. PLoS ONE. 2016;11(5). Ale A., Bruna J., Calls A., Karamita M., Haralambous S., Probert L. et al. Inhibition of the neuronal NFκB pathway attenuates bortezomib-induced neuropathy in a mouse model. NeuroToxicology. 2016;55: Badia J, Raspopovic S, Carpaneto J, Micera S, Navarro X. Spatial and Functional Selectivity of Peripheral Nerve Signal Recording With the Transversal Intrafascicular Multichannel Electrode (TIME).IEEE transactions on neural systems and rehabilitation engineering : a publication of the IEEE Engineering in Medicine and Biology Society. 2016;24(1). Santos D., Giudetti G., Micera S., Navarro X., Del Valle J. Focal release of neurotrophic factors by biodegradable microspheres enhance motor and sensory axonal regeneration in vitro and in vivo. Brain Research. 2016;1636: Rubio MA, Herrando-Grabulosa M, Vilches JJ, Navarro X. Involvement of sensory innervation in the skin of SOD1(G93A) ALS mice.journal of the peripheral nervous system : JPNS. 2016;21(2). Velasco R., Santos C., Soler G., Gil-Gil M., Pernas S., Galan M. et al. Serum micronutrients and prealbumin during development and recovery of chemotherapy-induced peripheral neuropathy. Journal of the Peripheral Nervous System. 2016;21(3): CIBERNED Annual Report / 188

189 Program 2 Group: Xavier Navarro Acebes Carboni C., Bisoni L., Carta N., Puddu R., Raspopovic S., Navarro X. et al. An integrated interface for peripheral neural system recording and stimulation: system design, electrical tests and invivo results. Biomedical Microdevices. 2016;18(2). Micera S, Raspopovic S, Petrini F, Carpaneto J, Oddo C, Badia J, Stieglitz T, Navarro X, Rossini PM, Granata G. On the use of intraneural transversal electrodes to develop bidirectional bionic limbs. In: Converging Clinical and Engineering Research on Neurorehabilitation II. Ibáñez J, González-Vargas J, Azorín J, Akay M, Pons J, editors. Biosystems & Biorobotics, Springer, 2017, vol 15, pp ISBN: Stieglitz T, Boretius T, Čvančara P, Guiraud D, Guiho T, Lopez-Alvarez VM, Navarro X. Biocompatibility and stability of transversal intrafascicularmultichannel electrodes TIME. In: Converging Clinical and Engineering Research on Neurorehabilitation II. Ibáñez J, González Vargas J, Azorín J, Akay M, Pons J, editors. Biosystems & Biorobotics, Springer, 2017, vol 15, pp ISBN: Yoshida K, Malec J, Comoglio C, Mosier K, Lontis R, Larsen K, Navarro X, Jensen W. Evaluation of the effect of sensory feedback on phantom limb pain in multi-center clinical trials. In: Converging Clinical and Engineering Research on Neurorehabilitation II. Ibáñez J, González Vargas J, Azorín J, Akay M, Pons J, editors. Biosystems & Biorobotics, Springer, 2017, vol 15, pp ISBN: Research projects Code: RD12/0019/0008 Title: RETIC de terapia celular. Principal Investigator: Isabel Fariñas (coordinador de red: Jose Maria Moraleda) CIBERNED s collaboration: Yes. CIBERNED groups: G102, G301, G113, G208, G105, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: 2015/01. Title: Terapia génica dirigida a neuregulinas para el tratamiento de la degeneración de las motoneuronas en la esclerosis lateral amiotrofica. Principal Investigator: Xavier Navarro Acebes. CIBERNED s collaboration: Yes. CIBERNED groups: G607, G113, G406, G407. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: RD12/0019/0016. Title: Red Terapia Celular: NEUROCEL-ELA. Nodo Valencia. Principal Investigator: José Manuel García Verdugo. CIBERNED s collaboration: Yes. CIBERNED groups: G113, G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: RTC Title: Optimización y desarrollo de un agente terapéutico basado en ácidos nucleicos para el tratamiento de la enfermedad de Huntington. Principal Investigator: Assumpció Bosch (G607)/Esther Pérez (G301). CIBERNED s collaboration: Yes. CIBERNED groups: G607, G301. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: ,92. Duration: Code: RTC Title: Desarrollo de agentes terapéuticos basados en ácidos nucleicos para el tratamiento de enfermedades neuromotoras y neuromusculares. Principal Investigator: Rubén López Vales. CIBERNED s collaboration: Yes. CIBERNED groups: G607, G609. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 189

190 Code: PI15/ Title: Estudio exploratorio sobre la etiopatogenia, biomarcadores de riesgo y mecanismos implicados en la regeneración de la neuropatía inducida por platinos. Principal Investigator: Jordi Bruna. CIBERNED groups: G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: FP Title: Natural sensory feedback for phantom limb pain modulation and therapy (EPIONE). Principal Investigator: Xavier Navarro Acebes. CIBERNED groups: G607. Type: European. Funding agency: Comisión Europea. Funding: Duration: Code: FP Title: Neurocontrolled bidirectional artificial upper limb and hand prosthesis (NEBIAS). Principal Investigator: Xavier Navarro Acebes. CIBERNED groups: G607. Type: European. Funding agency: Comisión Europea. Funding: Duration: Code: SAF Title: Reprogramación neuronal para promover los mecanismos endogenos de neuroprotección usando biología sintética en un modelo de degeneración retrógrada de motoneuronas. Principal Investigator: Caty Casas. CIBERNED groups: G607. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: IRP Research Grant P148. Title: Role of pro-resolving lipid mediators in SCI. Principal Investigator: Rubén Lopez-Vales. CIBERNED groups: G607. Type: International. Funding agency: International Foundation for Research in Paraplegia. Funding: Duration: Code: TV Title: Terapia génica dirigida a neuregulinas para el tratamiento de la esclerosis lateral amiotrófica. Principal Investigator: Xavier Navarro. CIBERNED groups: G607. Type: Private. Funding agency: Fundació La Marató-TV3. Funding: Duration: Code: PI15/ Title: Estudio molecular y tratamiento de la Mucopolisacaridosis tipo VII. Principal Investigator: Assumpcio Bosch. CIBERNED groups: G607. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: SAF R. Title: Participación de los receptores del aácido lisofosfatídico en la fisiopatología de la lesión medular. Principal Investigator: Rubén Lopez-Vales. CIBERNED groups: G607. Type: National. Funding agency: Ministerio de Economía y Competitividad. Funding: Duration: CIBERNED Annual Report / 190

191 Program 2 Group: Xavier Navarro Acebes PhD dissertations Author: Francisco J. González Pérez. Title: Functionalizing artificial nerve guides to promote regeneration and recovery after peripheral nerve injuries. Date: 1 February Supervisor: Xavier Navarro Acebes. Author: Daniel Santos Rojas. Title: Contributions to the development of a bio-electronic regenerative interface for the injured peripheral nerve Date: 21 July Supervisor: Xavier Navarro Acebes. Author: Isaac Francos Quijorna. Title: Activation of inflammatory resolution programs as a new therapeutic approach to promote neuroprotection after SCI. Date: 22 July Supervisor: Rubén López Vales. Author: Ariadna Arbat Plana. Title: Modulation of the stretch reflex arc to improve functional recovery after peripheral nerve injury. Date: 19 Septembre Supervisor: Esther Udina Bonet CIBERNED Annual Report / 191

192 GROUP José Ángel Obeso Inchausti 205 Principal Investigator José Ángel Obeso Inchausti Research team Francisco Molinet Dronda Researcher Fernando Alonso Frech Researcher Michele Dileone Researcher Stéphanie Etienne Researcher Raquel Márquez López Technician Raúl Martínez Fernández Researcher Francisco Javier Blesa de los Mozos PhD Cristina Calvo González Administrative assistant Iván Castela Muñoz Researcher Marta del Álamo de Pedro Researcher María Ledia Fernández Hdez. PhD Guillermo Foffani PhD Carmen Gasca Salas Researcher Frida Hernández Fernández Researcher Natalia López González del Rey PhD Centro Integral de Neurociencias AC (CINAC) Hospital Puerta del Sur y Universidad San Pablo CEU, Avenida Carlos V, nº Móstoles, Madrid (Spain) Tel.: jobeso.hmcinac@hmhospitales.com Michele Matarazzo Researcher Ignacio Obeso Martín PhD José Ángel Pineda Pardo PhD Rafael Rodríguez Rojas Researcher Álvaro Sánchez Ferro Researcher Inés Trigo Damas PhD Lydia Vela Desojo Researcher 2016 CIBERNED Annual Report / 192

193 Summary Dr. Obeso heads the movement disorders which is mainly focused on the study of Parkinson s disease (PD) from a multidisciplinary approach. The principal objective is the study of the onset and progression of the disease using basic and clinical experiments aiming to define the etiopathogenesis of the disease, optimizing the diagnosis and advancing in the treatment of neurodegenerative and neuropsiquiatric diseases. Onset of the disease: compensatory mechanisms and selective vulnerability So far, treatments for PD have not been successful and one of the reasons is because diagnosis tends to be delayed respect to the onset of the neurodegenerative process. At CINAC, patients are treated with magnetic stimulation (TMS) and neuroimaging directed to define the onset of the disease. Interestingly, PD is principally characterized by the progressive degeneration of dopaminergic neurons of the substantia nigra pars compacta (SNc), which are crucial during learning and habit acquisition. Additionally, since the dopaminergic loss is previous to the diagnosis it is assumed that there might be a series of compensatory mechanisms which delay the rise of the symptoms. At CINAC, in vivo neuronal activity recordings in PD animals models are used to define these mechanisms. Evolution and treatment of the disease HIFU (High Intensity Focused Ultrasound) is a novel technique which allows performing controlled and focused brain lesions without surgery. Thus, mobility and mortality and economic costs are greatly reduced compared to current surgical treatments. This technique has demonstrated efficacy and safety in the performance of thalamotomies aimed to cease essential tremor and PD tremor. At CINAC, the team uses this technique to improve the symptoms. All studies mentioned above are supplemented with neuroimaging studies using positron emission topography (PET) and magnetic resonance (MR) which help to determine potential biomarkers of the disease. Neuroinflammation and Parkinson s disease Several mechanisms are involved in the etiopathogeny and progression of PD and, neuroinflamation and glial activation have gain relevance among them. Studies with longterm disease patients and with animal models showing severe dopamine depletion reveal numerous evidences of a glial and astrocytes activation and high levels of proinflammation citokynes in SNc and striatum, which are related to the dopamine loss. At CINAC imaging in vivo studies in animal models are used in order to examine the relationship between the neuroinflamation processes and the dopamine loss in PD. Pharmacological treatment of Parkinson s disease and motor complications Dopamine replacement therapy has been largely used when treating PD. Both dopamine agonists and levodopa (LDOPA) are the main treatments. Even if quite successful in ameliorating the motor symptoms, this therapy is also associated not only with motor complications after long term use (involuntary movements: dyskinesias). At CINAC animal models showing progressive dopamine depletion are utilized in order to determine the molecular base and mechanisms involved in the onset of LDOPA dyskinesias (LID) using recording and optogenetic techniques which allow a more specific and selective neuronal modulation. Keywords Parkinson s disease, Substantia nigra pars compacta, Basal Ganglia, Dopamine, Vulnerability, Dyskinesias, Compensatory mechanisms Publications De la Casa-Fages B., Alonso-Frech F., Grandas F. Effect of subthalamic nucleus deep brain stimulation on balance in Parkinson s disease: A static posturographic analysis. Gait and Posture. 2017;52: Epub Ponsoda V., Martinez K., Pineda-Pardo J.A., Abad F.J., Olea J., Roman F.J. et al. Structural brain connectivity and cognitive ability differences: A multivariate distance matrix regression analysis. Human Brain Mapping. 2017;38(2): Epub CIBERNED Annual Report / 193

194 Humanes-Valera D, Foffani G, Alonso-Calviño E, Fernández-López E, Aguilar J. Dual Cortical Plasticity After Spinal Cord Injury.Cerebral cortex (New York, N.Y. : 1991) Contador I, Stern Y, Bermejo-Pareja F, Sánchez-Ferro Á, Benito-León J. Is educational attainment associated with increased risk of mortality in people with dementia?. A population-based study. Current Alzheimer research Di Lazzaro V., Pellegrino G., Capone F., Florio L., Dileone M., Cioni B. et al. Reduction of disease progression in a patient with amyotrophic lateral sclerosis after several years of epidural motor cortex stimulation. Brain Stimulation Garcia-Garcia D, Guridi J, Toledo JB, Alegre M, Obeso JA, Rodríguez-Oroz MC. Stimulation sites in the subthalamic nucleus and clinical improvement in Parkinson s disease: a new approach for active contact localization.journal of neurosurgery Giancardo L., Sanchez-Ferro A., Arroyo-Gallego T., Butterworth I., Mendoza C.S., Montero P. et al. Computer keyboard interaction as an indicator of early Parkinson s disease. Scientific Reports. 2016;6. Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Cerasa A., Obeso I., Dileone M., Quattrone A. Transcranial Non-Invasive Brain Stimulation in Parkinson s Disease Patients with Dyskinesias. Where is the Optimal Target?. Cerebellum. 2016; 1-3. Ligneul R., Obeso I., Ruff C.C., Dreher J.-C. Dynamical Representation of Dominance Relationships in the Human Rostromedial Prefrontal Cortex. Current Biology. 2016;26(23): Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. Singh A., Mewes K., Gross R.E., DeLong M.R., Obeso J.A., Papa S.M. Human striatal recordings reveal abnormal discharge of projection neurons in Parkinson s disease. Proceedings of the National Academy of Sciences of the United States of America. 2016;113(34): Calabresi P., Pisani A., Rothwell J., Ghiglieri V., Obeso J.A., Picconi B. Hyperkinetic disorders and loss of synaptic downscaling. Nature Neuroscience. 2016;19(7): Benito-Leon J., Louis E.D., Mato-Abad V., Dydak U., Alvarez-Linera J., Hernandez-Tamames J.A. et al. In vivo neurometabolic profiling in orthostatic tremor. Medicine (United States). 2016;95(37). Alonso-Calvino E., Martinez-Camero I., Fernandez-Lopez E., Humanes-Valera D., Foffani G., Aguilar J. Increased responses in the somatosensory thalamus immediately after spinal cord injury. Neurobiology of Disease. 2016;87: Aurtenetxe S., Garcia-Pacios J., Rio D., Lopez M.E., Pineda-Pardo J.A., Marcos A. et al. Interference impacts working memory in mild cognitive impairment. Frontiers in Neuroscience. 2016;10. Domingo-Santos A., Sepulveda M., Matarazzo M., Calleja-Castano P., Ramos-Gonzalez A., Saiz A. et al. Intravenous immunoglobulin therapy in a patient with anti-myelin oligodendrocyte glycoprotein-seropositive neuromyelitis optica. Clinical Neuropharmacology. 2016;39(6): Gasca-Salas C, Alonso A, González-Redondo R, Obeso JA. Coexisting Parkinson s and Wilson s Disease: Chance or Connection?. The Canadian journal of neurological sciences. Le journal canadien des sciences neurologiques Matarazzo M., Galan Sanchez-Seco V., Mendez-Guerrero A.J., Gata-Maya D., Domingo- Santos A., Ruiz-Morales J. et al. Drug-Related Eyelid Nystagmus: Two Cases of a Rare Clinical Phenomenon Related to Carbamazepine and Derivatives. Clinical Neuropharmacology CIBERNED Annual Report / 194

195 Program 2 Group: José Ángel Obeso Inchausti Ferros I., Mora M.J., Obeso I.F., Jimenez P., Martinez-Insua A. Relationship between the cranial base and the mandible in artificially deformed skulls. Orthodontics and Craniofacial Research. 2016;19(4): Blesa J, Trigo-Damas I, Quiroga-Varela A, Lopez- Gonzalez del Rey N. Parkinson s Disease- Associated Mutations Affect Mitochondrial Function. In: Mitochondrial Mechanisms of Degeneration and Repair in Parkinson s Disease. Buhlman, Lori M, editors. Springer, 2016, pp ISBN: Gasca-Salas C. Treatment of cognitive impairment in Parkinson disease: A practical approach. Medicina Clinica Hernandez L.F., Obeso I. A STOP signal to striatum mediated by globus pallidus: A new loop discovered. Movement Disorders. 2016;31(8): Martínez-Fernández R, Gasca-Salas C, Sánchez-Ferro A, Obeso JA. Actualización en la enfermedad de Parkinson. Revista Médica Clínica Las Condes. 2016; 27(3), Sanchez-Ferro A., Maetzler W. Advances in sensor and wearable technologies for Parkinson s disease. Movement Disorders. 2016;31(9): Hernandez L.F. An A-PLAuse to a new assay that unveils previously undetected alphasynucleinopathies. Movement Disorders. 2016;31(3):301-. Trigo-Damas I., Gonzalez del Rey N.L. Are the basal ganglia actually controlling movement or quite the opposite?. Movement Disorders. 2016;31(9): Gasca-Salas C., Masellis M., Khoo E., Shah B.B., Fisman D., Lang A.E. et al. Characterization of movement disorder phenomenology in genetically proven, familial frontotemporal lobar degeneration: A systematic review and meta-analysis. PLoS ONE. 2016;11(4). Dileone M., Ranieri F., Florio L., Capone F., Musumeci G., Leoni C. et al. Differential Effects of HRAS Mutation on LTP-Like Activity Induced by Different Protocols of Repetitive Transcranial Magnetic Stimulation. Brain Stimulation. 2016;9(1): Trigo-Damas I. Dopamine Receptors Direct the Plasticity Orchestra in the Motor Cortex. Movement Disorders. 2016;31(7):969-. Obeso I., Oliviero A., Jahanshahi M. Editorial: Non-invasive brain stimulation in neurology and psychiatry. Frontiers in Neuroscience. 2016;10. Crossman AR, Obeso JA. Functions of the basal ganglia-paradox or no paradox?. Movement disorders : official journal of the Movement Disorder Society. 2016;31(8). Garces P., Pereda E., Hernandez-Tamames J.A., Del-Pozo F., Maestu F., Angel Pineda-Pardo J. Multimodal description of whole brain connectivity: A comparison of resting state MEG, fmri, and DWI. Human Brain Mapping. 2016;37(1): Sanchez-Ferro A., Elshehabi M., Godinho C., Salkovic D., Hobert M.A., Domingos J. et al. New methods for the assessment of Parkinson s disease (2005 to 2015): A systematic review. Movement Disorders. 2016;31(9): Blesa J, Trigo-Damas I, Obeso JA. Parkinson s disease and thalamus: facts and fancy.the Lancet. Neurology. 2016;15(7). Martinez-Fernandez R., Pelissier P., Quesada J.-L., Klinger H., Lhommee E., Schmitt E. et al. Postoperative apathy can neutralise benefits in quality of life after subthalamic stimulation for Parkinson s disease. Journal of Neurology, Neurosurgery and Psychiatry. 2016;87(3): Benito-Leon J., Louis E.D., Manzanedo E., Hernandez-Tamames J.A., Alvarez-Linera J., Molina- Arjona J.A. et al. Resting state functional MRI reveals abnormal network connectivity in orthostatic tremor. Medicine (United States). 2016;95(29). del Rey N.L.-G., Blesa J. Selective connectivity of dopamine neurons projecting to the posterior striatum. Movement Disorders. 2016;31(3):298-. Herreros-Rodriguez J., Sanchez-Ferro A. Summertime dyskinesia-hyperpyrexia syndrome: The Dual Heat hypothesis. Clinical Neuropharmacology. 2016;39(4): CIBERNED Annual Report / 195

196 Obeso I., Cerasa A., Quattrone A. The effectiveness of transcranial brain stimulation in improving clinical signs of hyperkinetic movement disorders. Frontiers in Neuroscience. 2016;9(JAN). Martinez-Fernandez R., Schmitt E., Martinez-Martin P., Krack P. The hidden sister of motor fluctuations in Parkinson s disease: A review on nonmotor fluctuations. Movement Disorders. 2016;31(8): Vela L., Martinez Castrillo J.C., Garcia Ruiz P., Gasca-Salas C., Macias Macias Y., Perez Fernandez E. et al. The high prevalence of impulse control behaviors in patients with earlyonset Parkinson s disease: A cross-sectional multicenter study. Journal of the Neurological Sciences. 2016;368: Obeso J.A. The Movement Disorders journal 2016 and onward. Movement Disorders. 2016;31(1):1-2. Blesa J. Vive la difference! Dissecting the diversity of midbrain dopamine neurons. Movement Disorders. 2016;31(1): Research projects Code: SAF C 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. CIBERNED groups: G205. Type: International. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: Sin código. Title: Selective vulnerability, progression and synuclein toxicity in Parkinson`s disease. Principal Investigator: José Ángel Obeso Inchausti. CIBERNED groups: G205. Type: Private. Funding agency: BBVA. Funding: Duration: Code: MJFF Staff Initiated Title: Prion-like dissemination of synuclein pathology: a non-human primate study. Principal Investigator: Erwan Bezard (Universite de Bordeaux, France). CIBERNED s collaboration: Yes. CIBERNED groups: G109, G205. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: Code: FJCI Title: Programa Juan De La Cierva. Principal Investigator: José Ángel Pineda Pardo. CIBERNED groups: G205. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 196

197 Program 2 Group: José Ángel Obeso Inchausti Code: CD15/ Title: Contrato posdoctoral de perfeccionamiento Sara Borrell. Grupo habitual. Principal Investigator: Ignacio Obeso Martín. CIBERNED groups: G205. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: H2020-MSCA-IF-2014 ROSNPD Title: Marie Skłodowska-Curie Individual Fellowships. Principal Investigator: Ledia Fernández Hernández. CIBERNED groups: G205. Type: European. Funding agency: Comisión Europea. Funding: Duration: PhD dissertations Author: Carmen Gasca. Title: Estudio prospectivo del deterioro cognitivo en la enfermedad de Parkinson. Date: 8 February Supervisor: José Ángel Obeso Inchausti CIBERNED Annual Report / 197

198 GROUP Ana Pérez-Castillo 110 Principal Investigator Ana Pérez-Castillo Research team Ángel Santos Montes Full professor José A. Morales García PhD Elena Giné PhD Sandra Alonso Gil Technician Marina Sanz San Cristóbal Technician Ana Beltrán Arranz Student Adriana Escudero Pérez Student Instituto de Investigaciones Biomédicas (CSIC-UAM) Dpto. de Modelos Experimentales de Enfermedades Humanas c/arturo Duperier Madrid (Spain) Tel.: Fax: CIBERNED Annual Report / 198

199 Summary Research is focused on molecular biological approaches to study pathologies of the central nervous system. One major area of interest is the identification of new targets for the treatment of several CNS disorders, specifically Parkinson s disease, as well as in the development of proofof concept for novel drugs directed towards these targets. We have identified several genes involved in different brain disorders, including the nuclear receptor PPARγ, GSK-3, C/EBPβ, and phosphodiesterase 7 (PDE7). In this regard we have shown that ligands of PPARγ and new inhibitors of PDE7 and GSK-3 are potent anti-inflammatory and neuroprotective agents in different experimental models of brain diseases. Also, a main focus of the lab concerns research on neurogenesis and aging. We are currently working in the role of different new targets which can expand our understanding of the processes that lead to improved neurogenesis and that can be of use for a better understanding and new treatments of aging-related disorders. Keyword Neuroprotección, neuroinflamación, neurogénesis, Parkinson, PDE7, C / EBPβ, C3 Publications Morales-Garcia J.A., Echeverry-Alzate V., Alonso-Gil S., Sanz-Sancristobal M., Lopez-Moreno J.A., Gil C. et al. Phosphodiesterase7 Inhibition Activates Adult Neurogenesis in Hippocampus and Subventricular Zone In Vitro and In Vivo. Stem Cells Hernandez-Encinas E, Aguilar-Morante D, Morales-Garcia JA, Gine E, Sanz-SanCristobal M, Santos A et al. Complement component 3 (C3) expression in the hippocampus after excitotoxic injury: role of C/EBPβ.Journal of neuroinflammation. 2016;13(1). Estrada M., Herrera-Arozamena C., Perez C., Vina D., Romero A., Morales-Garcia J.A. et al. New cinnamic - N-benzylpiperidine and cinnamic - N,N-dibenzyl(N-methyl)amine hybrids as Alzheimer-directed multitarget drugs with antioxidant, cholinergic, neuroprotective and neurogenic properties. European Journal of Medicinal Chemistry. 2016;121: Estrada M., Perez C., Soriano E., Laurini E., Romano M., Pricl S. et al. New neurogenic lipoicbased hybrids as innovative Alzheimer s drugs with σ-1 agonism and β-secretase inhibition. Future Medicinal Chemistry. 2016;8(11): Research projects Code: 2015/03. Title: Perfiles metabólicos diferenciales en enfermedad de Parkinson. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED s collaboration: Yes. CIBERNED groups: G103, G110, G209. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: 2015 Code: SAF2014. Title: Papel del factor de transcripción CEBPβ en la enfermedad de Parkinson: Identificación y caracterización de los genes mediadores de su acción. Principal Investigator: Ana Pérez Castillo. CIBERNED groups: G110. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 199

200 Code: 2014 Target validation. Title: Effects of new agonists of melatonin receptors on neurodegeneration, inflammation, and neuroregeneration in Parkinson s disease. Principal Investigator: Ana Pérez Castillo. CIBERNED groups: G110. Type: International. Funding agency: Michael J. Fox Foundation. Funding: N.D. Duration: Code: Marie Skłodowska-Curie Actions. H2020-MSCA-ITN Title: Blood biomarker-based diagnostic tools for early-stage Alzheimer s disease. Principal Investigator: Ana Pérez Castillo. CIBERNED groups: G110. Other CIBER s collaboration: Yes (CIBERER). Type: European. Funding agency: Comisión Europea. Funding: ,52. Duration: Code: Marie Skłodowska-Curie Actions. H2020-MSCA-ITN Title: Neuroregenerative drug discovery. Principal Investigator: María Laura Bolognesi. CIBERNED groups: G110. Type: European. Funding agency: Comisión Europea. Funding: N.D. Duration: CIBERNED Annual Report / 200

201 GROUP Jordi Pérez Tur 209 Principal Investigator Jordi Pérez Tur CSIC Research team Fernando Cardona Serrate Researcher Concepción Rubio Granero Researcher Jacek Szymanski Researcher Àngels Albors Asensi Lab technician Jessica Valdivia Lab technician Guillermo Rodríguez Casero Lab technician Unitat de Genètica Molecular Institut de Biomedicina de València-CSIC c/jaume Roig, Valencia (Spain) Tel.: Fax: CIBERNED Annual Report / 201

202 Summary During this year, we have focused on the study of genetic factors related to diseases of the spectrum Amyotrophic Lateral Sclerosis Frontotemporal Dementia and Parkinson s disease, taking advantage of the synergies with other groups of CIBERNED through DEGESCO. As for work in progress, throughout 2016 we have been studying the genetic factors involved in the onset of dementia in Parkinson s disease, the characterization of variants or alterations in DNA related to the appearance of a Familial form of primary lateral sclerosis as well as in the definition of the mutation that causes a familial form of Progressive Supranuclear Palsy. Likewise, we have continued the functional characterization of variants that may be related to the risk of Parkinson s disease as well as variants that modify the expression of genes related to this disease. Keywords Alzheimer, Parkinson, Neurodegeneration, Neurogenetics, ALS Publications Cardona F, Pérez-Tur J. Other proteins involved in Parkinson s disease and related disorders. Current protein & peptide science Research projects Code: 2015/03. Title: Perfiles metabólicos diferenciales en enfermedad de Parkinson. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED s collaboration: Yes. CIBERNED groups: G103, G110, G209. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: 2015 Code: SAF R. Title: A la caza de genes esquivos: ELA, PSP y epilepsia. Principal Investigator: Jordi Pérez Tur. CIBERNED groups: G209. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 202

203 GROUP José Antonio del Río Fernández 114 Principal Investigator José Antonio del Río Fernández Research projects Rosalina Gavín Marín Lecturer Vanessa Gil Fernández Research fellow Arnau Hervera Abad Research fellow Andreu Matamoros Anglès Ágata Mata Rodríguez Laura Urrea Zazurca Laila Lidón Gil Ana López Mengual Francina Mesquida Veny Master student Miriam Segura Feliú Lab technician Molecular and Cellular Neurobiotechnology Institute for Bioengineering of Catalonia (IBEC) Baldiri Reixac Barcelona, Catalunya (Spain) Tel.: / jadelrio@ub.edu jadelrio@ibecbarcelona.eu 2016 CIBERNED Annual Report / 203

204 Summary Our research interests are focused on three main aspects of Neurodegeneration: Reelin and rapid progressive dementia Reelin is an extracellular glycoprotein involved in key cellular processes in developing and adult nervous system, including regulation of neuronal migration, synapse formation and plasticity. Most of these roles are mediated by the intracellular phosphorylation of Dab1, an intracellular adaptor molecule, in turn mediated by binding Reelin to its receptors. Altered expression and glycosylation patterns of Reelin in cerebrospinal and cortical extracts have been reported in Alzheimer s disease. However, putative changes in Reelin are not described in natural prionopathies or experimental models of prion infection or toxicity. With this is mind, we determined that Reelin protein and mrna levels increased in CJD human samples and in mouse models of human prion disease in contrast to murine models of prion infection (Mata et al., Mol Neurobiol Oct 10). However, changes in Reelin expression appeared only at late terminal stages of the disease, which prevents their use as an efficient diagnostic biomarker. In addition, increased Reelin in CJD and in in vitro models does not correlate with Dab1 phosphorylation, indicating failure in its intracellular signaling. Overall, these findings widen our understanding of the putative changes of Reelin in neurodegeneration, particularly in rapid progressive dementias. α-synuclein spreading in Parkinson s disease The cellular prion protein, encoded by the gene PRNP, has been reported as receptor of β-amyloid (reviewed in Del Río and Gavín 2016). Their interaction is mandatory for neurotoxic effects of β-amyloid oligomers. We aimed to explore whether the cellular prion protein participates in the spreading of α-synuclein (Figure 1). Results demonstrate that PRNP expression is not mandatory for α-synuclein spreading (Urrea et al., Mol Neurobiol Feb 22). However, although the pathological spreading of α-synuclein can take place in the absence of PRNP, α-synuclein fibrils bind strongly on PRNP expressing cells, suggesting a role modulating the effect of α-synuclein fibrils. In fact, increased levels of PRNP enhance the spreading of α-synuclein in living mice. Taken together we demonstrate that the cellular prion protein is a receptor/binding partner of α-synuclein. Development of new lab on a chip devices for neurobiological research We recently developed a new device able to reproduce axon lesioning in vitro in a single chip. Current experiments of our group in collaboration with groups of IBEC and Cinerned aimed at developing new lab on chip devices to mimics and modulate neurodegeneration. For example: cortico-spinal chips to develop genetic studies; molecular gradient generation for migrating neurons and in silico 3D modeling for neurodegenerative diseases (Parkinson s and Alzheimer s diseases chip). At this respect, we aimed to mimic neuronal pathways of relevance in neurodegeneration. First experiments are developed to reconstitute in vitro the connections between cortex-striatumsubstantia nigra. Keywords P-Tau, amyloid, PrPc, axonal regeneration, cell therapy Publications Frau-Mendez M.A., Fernandez-Vega I., Ansoleaga B., Blanco Tech R., Carmona Tech M., Antonio del Rio J. et al. Fatal familial insomnia: Mitochondrial and protein synthesis machinery decline in the mediodorsal thalamus. Brain Pathology Garcia-Esparcia P, Koneti A, Rodríguez-Oroz MC, Lago B, Del Rio JA, Ferrer I. Mitochondrial activity in the frontal cortex area 8 and angular gyrus in Parkinson s disease and Parkinson s disease with dementia.brain pathology (Zurich, Switzerland) Gutierrez-Franco A., Eixarch H., Costa C., Gil V., Castillo M., Calvo-Barreiro L. et al. Semaphorin 7A as a Potential Therapeutic Target for Multiple Sclerosis. Molecular Neurobiology. 2016; CIBERNED Annual Report / 204

205 Program 2 Group: José Antonio del Río Fernández Del Río JA, Gavín R. Functions of the cellular prion protein, the end of Moore s law, and Ockham s razor theory.prion Ansoleaga B., Garcia-Esparcia P., Llorens F., Hernandez-Ortega K., Carmona M., Del Rio J.A. et al. Altered mitochondria, protein synthesis machinery, and Purine metabolism are molecular contributors to the pathogenesis of Creutzfeldt-Jakob disease. Journal of Neuropathology and Experimental Neurology. 2016;75(8): Mata A., Urrea L., Vilches S., Llorens F., Thune K., Espinosa J.-C. et al. Reelin Expression in Creutzfeldt- Jakob Disease and Experimental Models of Transmissible Spongiform Encephalopathies. Molecular Neurobiology. 2016; Tomas-Roig J, Piscitelli F, Gil V, Del Río JA, Moore TP, Agbemenyah H et al. Social defeat leads to changes in the endocannabinoid system: An overexpression of calreticulin and motor impairment in mice.behavioural brain research. 2016;303. Requena J.R., Kristensson K., Korth C., Zurzolo C., Simmons M., Aguilar-Calvo P. et al. The Priority position paper: Protecting Europe s food chain from prions. Prion. 2016;10(3): Research projects Code: 392/U/2014. Title: Role of the cellular prion protein as cross-talk protein between alpha-syn/ LRRK2 and p-tau in sporadic and familiar Parkinson s disease. Principal Investigator: José Antonio del Río (Fundacio Institut de Bioenginyeria de Catalunya). CIBERNED s collaboration: Yes. CIBERNED groups: G109, G114. Type: Private. Funding agency: Fundació la Marató TV3. Funding: Duration: Code: Marató TV3-6. Title: Paper de la PrPc com a cross-talk protein entre alpha-sinucleina/lrrk2 i p-tau en la malaltia de Parkinson de tipus esporàdic i/o familiar. Principal Investigator: José Antonio del Río. CIBERNED groups: G114. Type: Private. Funding agency: Fundació Marato TV3. Funding: ,25. Duration: Code: AGL REDT. Title: Red nacional de priones. Principal Investigator: José Antonio del Río CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G609, G207. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad.F Funding: Duration: Code: PI2016/02. Title: Monitoring the onset and evolution of neuronal dysfunctions in propagative neural disorders using microfluidic devices and translational approaches. Principal Investigator: José Antonio del Río. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G401, G201. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: CINERNED 2014/02. Title: Mecanismos epigenéticos implicados en la etiología y progresión de las demencias neurodegenerativas rápidamente progresivas Principal Investigator: Miguel Calero. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G601. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 205

206 GROUP Manuel Rodríguez Díaz 206 Principal Investigator Manuel Rodríguez Díaz Research team Tomás González Hernández Research staff Magdalena Sabaté Bel Research staff Alberto Sánchez Fernández Research staff Ingrid Morales Pérez Postdoctoral fellow Clara Rodríguez Sabaté Fernando Isidro Montón Álvarez Research staff Jesús Norellis Lorenzo Brito Research staff Departamento de Fisiología Facultad de Medicina Universidad de La Laguna La Laguna, Canarias (Spain) Tel.: Fax: CIBERNED Annual Report / 206

207 Summary Increasing evidence suggests that the dopaminergic degeneration which characterizes Parkinson s disease starts in the striatal dopamine terminals and progresses retrogradely to the body of dopamine cells in the substantia nigra. The role of striatal astrocytes in the striatal initiation of the dopaminergic degeneration is little known. We studied the astrocytic response to the dopaminergic denervation of the striatum. The injection of 6-hydroxydopamine (25 μg) in the lateral ventricle of adult rats induced a fast (4 hours) and selective (unaccompanied by unspecific lesions of striatal tissue or microgliosis) degeneration of the dopaminergic innervation of the striatum which was followed by a selective astrocytosis unaccompanied by microgliosis. This astrocytosis was severe and had a specific profile which included some (e.g. upregulation of GFAP, GS, S100β, NDRG2, vimentin) but not all (e.g. astrocytic proliferation or differentiation from NG2 cells, astrocytic scars, microgliosis) the characteristics observed after the non-selective lesion of the striatum. This astrocytosis is similar to those observed in the parkinsonian striatum. In these rats, dopaminergic debris were accumulated within spheroids (free-spheroids) which retained some proteins of dopaminergic neurons (e.g., tyrosine hydroxylase, the dopamine transporter protein, and APP) but not others (e.g., α-synuclein). Free-spheroids showed the initial (LC3- autophagosomes) but not the late (Lamp1/ Lamp2-lysosomes and activated Caspase 6) components of autophagia (incomplete autophagia), preparing their autophagosomes for an external phagocytosis (accumulation of phosphatidylserine). Free-spheroids were penetrated by astrocyte processes (fenestratedspheroids) which made them immunoreactive for GFAP and S100β, and which had the elements needed to continue the debris degradation (caspase 6 and Lamp1). Finally, proteins normally found in neurons (TH, DAT and α-synuclein) were observed within astrocytes 2-5 days after the dopaminergic degeneration, suggesting that the intracellular contents of degenerated cells had been transferred to astrocytes. Taken together, present data suggest phagocytosis as a physiological role of striatal astrocytes, a role which could be critical for cleaning striatal debris during the initial stages of Parkinson s disease. Our group is collaborating in the ips project with other groups belonging to CIBERNED who facilitate patient samples and the differentiation of astrocytic cells. By using a procedure developed to identify and to get representative BOLD-signals of the main basal ganglia from magnetic resonance images, the main interactions of these centers were studied in the human brain. The partial correlation methods showed a circular cortico-subcortical interaction which was globally similar in humans to that previously found in animals with other methods. This procedure was used to analyze the functional connectivity of the intralaminar thalamic centers with the main basal ganglia circuits. Data analysis with Multiple Linear Regression showed relevant non-linear components in the basal ganglia interactions. Thus, a new non-linear method based on the Multiple Correspondence Analysis was also applied to basal ganglia interactions. This is a multivariate method which may identify massive interacting networks with an unsupervised data-driven procedure. This method generates a self-organizing clustering of centers according to their interactions which is similar to that generated by Independent Component Analysis, but which may be particularly suitable when non-linear interactions are being studied. The Multiple Correspondence Analysis isolated six networks within the cortico-subcortical motor loop of basal ganglia. The activity of these networks is now being studied in Parkinson s disease. 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 phosforilation and ubiquitinization of the dopamine transporter. Keywords Astrocytes, IPs, dopaminergic degeneration, dopamine transporter, dopaminergic vulnerability, functional neuroimaging 2016 CIBERNED Annual Report / 207

208 Publications Morales I., Sanchez A., Rodriguez-Sabate C., Rodriguez M. The astrocytic response to the dopaminergic denervation of the striatum. Journal of Neurochemistry Rodriguez-Sabate C., Sabate M., Llanos C., Morales I., Sanchez A., Rodriguez M. The functional connectivity in the motor loop of human basal ganglia. Brain Imaging and Behavior. 2016; Barroso-Chinea P., Cruz-Muros I., Afonso-Oramas D., Castro-Hernandez J., Salas-Hernandez J., Chtarto A. et al. Long-term controlled GDNF over-expression reduces dopamine transporter activity without affecting tyrosine hydroxylase expression in the rat mesostriatal system. Neurobiology of Disease. 2016;88: CIBERNED Annual Report / 208

209 GROUP Eduard Tolosa Sarró 207 Principal Investigator Eduard Tolosa Sarró Full professor Research team María José Martí Doménech Consultant Francesc Valldeoriola i Serra Consultant Esteban Muñoz Martínez Consultant Joan Santamaría Cano Consultant Alejandro Iranzo de Riquer Consultant Carles Gaig Specialist Yaroslau Compta Hinrjy Specialist Mario Ezquerra Trabalón Researcher Rubén Fernández-Santiago Researcher Mónica Serradell Eroles Carme Junqué Plaja Full professor Alicia Garrido Pla Manuel Fernández Sánchez Lab technician Laura Maragall Clerk Institut de Neurociencies Unitat de Parkinson i Trastorns Anormals del Moviment c/villarroel 171, esc 8-4 planta Barcelona (España) Tel.: Fax: etolosa@clinic.ub.es 2016 CIBERNED Annual Report / 209

210 Summary We have continued our work on prodromal Parkinson disease (PD) in high risk cohorts (idiopathic RBD and PD non-manifesting LRRK2 mutation carriers) by performing clinical and wet biomarkers studies (Mirelman et al. Mov Disord 2016; Vilas et al. Mov Disord 2016; Brokmann et al. J Neuroinflamation 2016; Brokmann et al. Neurology 2016), and neuroimaging markers (Marzi et al. Ann Neurol 2016; Marzi et al. Mov Disorders 2016). We have reported that high mitochondrial DNA content in CSF distinguishes idiopathic from LRRK2-associated PD suggesting that different biochemical pathways underlie neurodegeneration in these two disorders (Podlesniy et al. Neurobiol Dis 2016). irbd has been identified as the most specific predictor of PD since most patients with irbd eventually develop PD (Iranzo et al. Lancet Neurology 2016, Fernádez-Arcos et al. Sleep 2016). The evidence is shown in the identification of deposits of alpha-synuclein in the submandibular gland of living subjects with IRBD (Vilas et al. Lancet Neurol, 2016) and abnormal substantia nigra nigrosome density (De Marzi et al. Ann Neurol 2016) in living subjects. We have identified another disorders that may mimic the clinical symptomatology of RBD named periodic limb movements in sleep (Gaig et al. Sleep 2016) and overlap parasomnia (Matos et al Sleep Med). Even though RBD can occur in LRRK2 mutations inducing PD, we have found that these LRRK2 mutations are not detected in IRBD (Fernández- Santiago et al. Neurology 2016). In addition, we have reported the presence of different genetic risk factors related to PD which are also associated with irbd such as single nucleotide polymorphism in the MAPT gene (Fernández- Santiago et al. Neurol Genet 2016). We have also described the polysomnographyc features of irbd (Antelmi et al. Sleep Med Rev 2016), and the evolution of cognition in manifested PD with associated RBD (Chaiue et al. Parkinsonism Rel Disord 2016). We have consolidated the line of research in CSF and imaging biomarkers in PD related dementia (Compta et al. J Neurol Sci 2016), as well as clinico-pathological correlations in Parkinson s disease and other parkinsonisms (Compta et al: Neuropathol Appl Neuropathol 2016 a & b). The network collaborations with international consortiums has resulted in new publications as well (COSTdystonia: Valadas et al; Eur J Neurol 2016 & Neurotox res 2016; UK study on apomorphine and amyloid pathology in PD: Yarnall et al. Mov Dis 2016; genome wide analyses in dementia with Lewy bodies: Guerreiro R. Neurobiol Aging 2016; genome wide association study in MSA: Sailer et al. Neurology 2016). As part of the Catalan Multiple System Atrophy (MSA), cross-sectional analyses of cerebrospinal fluid markers are being undertaken (abstract,international Congress of Parkinson s Disease and Movement Disorders Society, Vancouver, 2017), as well as connectivity MRI studies in typical parkinsonisms. In the line of research of MRI studies, we identified three patterns of cortical thinning in non-demented PD patients: one with parieto occipital atrophy, another with fonto-medial and coccipital atrophy and a third without atrophy (Uribe et al, Mov Disord 2016). Moreover in asymptomatic LRRK2 mutation carriers we found that from MRI resting-state acquisition could be observed increased and decreased functional connectivity in the nigro-striatal circuitry (Vilas et al: Mov Disord 2016). Studies on experimental therapeutics have included reports on targeting of the subthalamic nucleus and fast track care in deep brain stimulation surgery (Vesper et al. World Neurosurg 2016; Martin N et al. Neurocirugia 2016) and long term results of levodopa carbidopa gel infusions in advanced PD (Valldeoriola et al. Neurodegen Dis Manag 2017). Keywords Neuroimaging and cognition, biological markers, pre-motor diagnosis, non-motor symptoms, motor complications, genetic variants, sleep disorders, experimental therapy Publications Brockmann K, Schulte C, Schneiderhan-Marra N, Apel A, Pont-Sunyer C, Vilas D et al. Inflammatory profile discriminates clinical subtypes in LRRK2-associated Parkinson s disease. European journal of neurology. 2017;24(2). Epub Giri A., Mok K.Y., Jansen I., Sharma M., Tesson C., Mangone G. et al. Lack of evidence for a role of genetic variation in TMEM230 in the risk for Parkinson s disease in the Caucasian population. Neurobiology of Aging. 2017;50:167.e e13. Epub CIBERNED Annual Report / 210

211 Program 2 Group: Eduard Tolosa Sarró Hamel W., Koppen J.A., Alesch F., Antonini A., Barcia J.A., Bergman H. et al. Targeting of the Subthalamic Nucleus for Deep Brain Stimulation: A Survey Among Parkinson Disease Specialists. Tell-Marti G, Puig-Butille JA, Potrony M, Plana E, Badenas C, Antonell A et al. A Common Variant in the MC1R Gene (p.v92m) is associated with Alzheimer s Disease Risk.Journal of Alzheimer s disease : JAD. 2017;. Epub Kun-Rodrigues C, Ross OA, Orme T, Shepherd C, Parkkinen L, Darwent L et al. Analysis of C9orf72 repeat expansions in a large international cohort of dementia with Lewy bodies.neurobiology of aging Dangla-Valls A., Molinuevo J.L., Altirriba J., Sanchez-Valle R., Alcolea D., Fortea J. et al. CSF microrna Profiling in Alzheimer s Disease: a Screening and Validation Study. Molecular Neurobiology. 2016; 1-8. Padilla N, Fransson P, Donaire A, Figueras F, Arranz A, Sanz-Cortés M et al. Intrinsic Functional Connectivity in Preterm Infants with Fetal Growth Restriction Evaluated at 12 Months Corrected Age.Cerebral cortex (New York, N.Y. : 1991) Sailer A., Scholz S.W., Nalls M.A., Schulte C., Federoff M., Price T.R. et al. A genome-wide association study in multiple system atrophy. Neurology. 2016;87(15): Dauvilliers Y, Gaig C, Barateau L, Graus F, Iranzo A, Lopez R et al. Absence of NMDA receptor antibodies in the rare association between Type 1 Narcolepsy and Psychosis.Scientific reports. 2016;6. Valadas A., Contarino M.-F., Albanese A., Bhatia K.P., Falup-Pecurariu C., Forsgren L. et al. Management of dystonia in Europe: A survey of the European network for the study of the dystonia syndromes. European Journal of Neurology Yarnall A.J., Lashley T., Ling H., Lees A.J., Coleman S.Y., O Sullivan S.S. et al. Apomorphine: A potential modifier of amyloid deposition in Parkinson s disease?. Movement Disorders. 2016;31(5): Mirelman A., Bernad-Elazari H., Thaler A., Giladi-Yacobi E., Gurevich T., Gana-Weisz M. et al. Arm swing as a potential new prodromal marker of Parkinson s disease. Movement Disorders. 2016;31(10): Nalls MA, Keller MF, Hernandez DG, Chen L, Stone DJ, Singleton AB et al. Baseline genetic associations in the Parkinson s Progression Markers Initiative (PPMI).Movement disorders : official journal of the Movement Disorder Society. 2016;31(1). Santos-Garcia D., Mir P., Cubo E., Vela L., Rodriguez-Oroz M.C., Marti M.J. et al. COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global -clinical evaluations, serum biomarkers, genetic studies and neuroimaging- prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression. BMC Neurology. 2016;16(1). Marras C., Alcalay R.N., Caspell-Garcia C., Coffey C., Chan P., Duda J.E. et al. Motor and nonmotor heterogeneity of LRRK2-related and idiopathic Parkinson s disease. Movement Disorders Ferreira D., Bartres-Faz D., Nygren L., Rundkvist L.J., Molina Y., Machado A. et al. Different reserve proxies confer overlapping and unique endurance to cortical thinning in healthy middle-aged adults. Behavioural Brain Research. 2016;311: Aguilar F., Cisternas A., Montserrat J.M., Avila M., Torres-Lopez M., Iranzo A. et al. Effect of Nasal Continuous Positive Pressure on the Nostrils of Patients with Sleep Apnea Syndrome and no Previous Nasal Pathology. Predictive Factors for Compliance. Archivos de Bronconeumologia. 2016;52(10): Santos-García D, Mir P, Cubo E, Vela L, Rodríguez-Oroz MC, Martí MJ et al. Erratum to: COPPADIS-2015 (COhort of Patients with PArkinson s DIsease in Spain, 2015), a global--clinical evaluations, serum biomarkers, genetic studies and neuroimaging--prospective, multicenter, non-interventional, long-term study on Parkinson s disease progression.bmc neurology. 2016;16. Martin N., Valero R., Hurtado P., Gracia I., Fernandez C., Rumia J. et al. Experience with Fast track postoperative care after deep brain stimulation surgery. Neurocirugia. 2016;27(6): Sansa G., Gavalda A., Gaig C., Monreal J., Ercilla G., Casamitjana R. et al. Exploring the presence of narcolepsy in patients with schizophrenia. BMC Psychiatry. 2016;16(1) CIBERNED Annual Report / 211

212 Antelmi E., Ferri R., Iranzo A., Arnulf I., Dauvilliers Y., Bhatia K.P. et al. From state dissociation to status dissociatus. Sleep Medicine Reviews. 2016;28:1-13. Brockmann K., Apel A., Schulte C., Schneiderhan-Marra N., Pont-Sunyer C., Vilas D. et al. Inflammatory profile in LRRK2-associated prodromal and clinical PD. Journal of Neuroinflammation. 2016;13(1). Cosin-Tomas M., Antonell A., Llado A., Alcolea D., Fortea J., Ezquerra M. et al. Plasma mir- 34a-5p and mir-545-3p as Early Biomarkers of Alzheimer s Disease: Potential and Limitations. Molecular Neurobiology. 2016; De Marzi R., Seppi K., Hogl B., Muller C., Scherfler C., Stefani A. et al. Loss of dorsolateral nigral hyperintensity on 3.0 tesla susceptibility-weighted imaging in idiopathic rapid eye movement sleep behavior disorder. Annals of Neurology. 2016;79(6): Tell-Martí G, Anton Puig-Butille J, Potrony M, Badenas C, Milà M, Malvehy J et al. Reply to Letter to Editor ANA Lack of association between MC1R variants and Parkinson s disease in European descent by Jiang Q and Liu G.Annals of neurology Podlesniy P., Vilas D., Taylor P., Shaw L.M., Tolosa E., Trullas R. Mitochondrial DNA in CSF distinguishes LRRK2 from idiopathic Parkinson s disease. Neurobiology of Disease. 2016;94: Vilas D., Iranzo A., Tolosa E., Aldecoa I., Berenguer J., Vilaseca I. et al. Assessment of α-synuclein in submandibular glands of patients with idiopathic rapid-eye-movement sleep behaviour disorder: A case-control study. The Lancet Neurology Fabbri M., Leodori G., Fernandes R.M., Bhidayasiri R., Marti M.J., Colosimo C. et al. Neutralizing Antibody and Botulinum Toxin Therapy: A Systematic Review and Meta-analysis. Neurotoxicity Research. 2016;29(1): Vilas D., Shaw L.M., Taylor P., Berg D., Brockmann K., Aasly J. et al. Cerebrospinal fluid biomarkers and clinical features in leucine-rich repeat kinase 2 (LRRK2) mutation carriers. Movement Disorders Salat D., Noyce A.J., Schrag A., Tolosa E. Challenges of modifying disease progression in prediagnostic Parkinson s disease. The Lancet Neurology Fraser K.B., Moehle M.S., Alcalay R.N., West A.B., E. Tolosa, J.F. Martí Massó. Urinary LRRK2 phosphorylation predicts parkinsonian phenotypes in G2019S LRRK2 carriers. Neurology. 2016;86(11): Cerquera C., Rumia J., Herrera J.M., Moreno V., Bargallo N., Valldeoriola F. A single case report of MR-guided focused ultrasound thalamotomy for tremor in fragile X-associated tremor/ ataxia. Parkinsonism and Related Disorders. 2016;28: Fernandez-Santiago R., Iranzo A., Gaig C., Serradell M., Fernandez M., Tolosa E. et al. Absence of LRRK2 mutations in a cohort of patients with idiopathic REM sleep behavior disorder. Neurology. 2016;86(11): Compta Y, Ramos-Campoy O, Grau-Rivera O, Colom-Cadena M, Clarimón J, Martí MJ et al. Conjoint FTLD-FUS of the neuronal intermediate filament inclusion disease type, progressive supranuclear palsy and alzheimer s pathology presenting as parkinsonism with early falls and late hallucinations, psychosis and dementia.neuropathology and applied neurobiology Vilas D., Segura B., Baggio H.C., Pont-Sunyer C., Compta Y., Valldeoriola F. et al. Nigral and striatal connectivity alterations in asymptomatic LRRK2 mutation carriers: A magnetic resonance imaging study. Movement Disorders Gaig C, Iranzo A, Pujo M, Perez H, Santamaria J. Periodic Limb Movements during Sleep Mimicking REM Sleep Behavior Disorder.Sleep Solla P, Grau O, Gelpi E, Marrosu F, Martí MJ. Pisa syndrome in a patient with pathologically confirmed Parkinson s disease.neuropathology and applied neurobiology Penades R., Pujol N., Catalan R., Masana G., Garcia-Rizo C., Bargallo N. et al. Cortical thickness in regions of frontal and temporal lobes is associated with responsiveness to cognitive remediation therapy in schizophrenia. Schizophrenia Research. 2016;171(1-3): CIBERNED Annual Report / 212

213 Program 2 Group: Eduard Tolosa Sarró Vilas D, Muxi A, Tolosa E, Compta, Y. Enfermedad de Parkinson asociada a mutación del gen LRRK2, que simula atrofia multisistémica por la combinación de parkinsonismo, disautonomía precoz y gammagrafía cardíaca normal. Revista de neurología. 2016; 63(2), Iranzo A., Santamaria J., Tolosa E. Idiopathic rapid eye movement sleep behaviour disorder: Diagnosis, management, and the need for neuroprotective interventions. The Lancet Neurology. 2016;15(4): Sierra M., Gelpi E., Marti M.J., Compta Y. Lewy- and Alzheimer-type pathologies in midbrain and cerebellum across the Lewy body disorders spectrum. Neuropathology and Applied Neurobiology. 2016;42(5): Valldeoriola F, Grandas F, Santos-García D, Regidor I, Catalán MJ, Arbelo JM et al. Longterm effectiveness of levodopa-carbidopa intestinal gel in 177 Spanish patients with advanced Parkinson s disease.neurodegenerative disease management. 2016;6(4). Vilas D, Muxi A, Tolosa E, Compta Y. [Parkinson s disease associated to mutation of the LRRK2 gene, mimicking multisystemic atrophy due to the combination of parkinsonism, early-onset dysautonomia and normal cardiac scintigraphy].revista de neurologia. 2016;63(2). Uribe C., Segura B., Baggio H.C., Abos A., Marti M.J., Valldeoriola F. et al. Patterns of cortical thinning in nondemented Parkinson s disease patients. Movement Disorders. 2016;31(5): Fernandez-Arcos A., Iranzo A., Serradell M., Gaig C., Santamaria J. The clinical phenotype of idiopathic rapid eye movement sleep behavior disorder at presentation: A study in 203 consecutive patients. Sleep. 2016;39(1): Matos N., Iranzo A., Gaig C., Santamaria J. Video-polysomnographic documentation of nonrapid eye movement sleep parasomnia followed by rapid eye movement sleep behavior disorder: a parasomnia overlap disorder?. Sleep Medicine. 2016;23: Compta Y., Buongiorno M., Bargallo N., Valldeoriola F., Munoz E., Tolosa E. et al. White matter hyperintensities, cerebrospinal amyloid-β and dementia in Parkinson s disease. Journal of the Neurological Sciences. 2016;367: Research projects Code: PI2016/06. Title: Identificación de vías fisiopatológicas y biomarcadores candidatos en la fase prediagnóstica de la enfermedad de Parkinson. Principal Investigator: Miquel Vila Bover. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G601, G207, G410. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: Code: 2014\FTPGB-LAGUNA. Title: Estudio del perfil transcripcional en subgrupos de pacientes prodrómicos con enfermedad de Parkinson. Principal Investigator: Ariadna Laguna Tuset. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G207. Type: Private. Funding agency: Fundación Tatiana Pérez de Guzmán el Bueno. Funding: Duration: Code: COURAGE - PD. Title: Comprehensive unbiased risk factor assessment for genetics and environment in Parkinson s disease. Principal Investigator: Thomas Gasser. CIBERNED groups: G207. Type: International. Funding agency: JPND. Funding: Duration: Code: N.D. Title: The Parkinson s Progression Markers Initiative (PPMI). Principal Investigator: Eduard Tolosa. CIBERNED groups: G207. Type: International. Funding agency: The Michael J. Fox Foundation. Funding: Duration: CIBERNED Annual Report / 213

214 Code: AGL REDT. Title: Red nacional de priones. Principal Investigator: José Antonio del Río. CIBERNED s collaboration: Yes. CIBERNED groups: G114, G509, G503, G609, G207. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: 288/C/2014. Title: Catalan Network of Multiple System Atrophy: Biomarkers and Pathophysiology. Principal Investigator: M.j. Marti (Hospital Clinic-IDIBAPS). CIBERNED s collaboration: Yes. CIBERNED groups: G109, G207. Type: Private. Funding agency: Fundació La Marató de TV3. Funding: Duration: Code: LRRK2 Resource RFA Title: Alpha-synuclein and other biomarkers in biological samples of LRKK2 PD. Principal Investigator: Eduard Tolosa. CIBERNED s collaboration: Yes. CIBERNED groups: G410, G207. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: Code: PI Title: Mitochondrial DNA copy number in LRRK2 fibroblasts. Principal Investigation: Eduard Soriano. CIBERNED s collaboration: Yes. CIBERNED groups: G410, G207. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: CIBERNED Annual Report / 214

215 GROUP Carlos Vicario Abejón 108 Principal Investigator Carlos Vicario Abejón CSIC senior scientist Research team Eva Díaz Guerra Postdoctoral fellow Eva Rodríguez Traver Elena Moreno Jiménez Master student Carlos Omar Oueslati Morales Master student Mª José Román Laboratory technician Pablo Fernández Rodríguez Master student Manuel Antonio Oria Muriel Master student CSIC Avenida Doctor Arce Madrid (Spain) Tel.: Fax: CIBERNED Annual Report / 215

216 Summary We have obtained induced pluripotent stem cells (ips cells or ipscs) by reprogramming fibroblasts from Alzheimer s disease patients carrying the epsilon 3 and epsilon 4 alleles of APOE gene or a mutation in the Presenilin gene. Thanks to this achievement, we are participating in ADAPTED, a European project that got financed in the call Innovative Medicine Initiative H2020-JTI- IMI Carlos Vicario is the lider of work package-2. Our results obtained in ips cells and ips-derived neurons from Parkinson s disease patients were presented during the IV International Congress on Research and Innovation in Neurodegenerative Diseases (Vicario et al., 2016). We have published an article in the journal Stem Cells reporting the role and mechanisms of action of IGF-I growth factor, specifically that synthetized in the brain, during postnatal-adult neurogenesis in the hippocampal dentate gyrus (Nieto-Estévez et al., 2016a). Moreover, we have critically revised IGF-I functions on neurogenesis and synaptogenesis (Nieto-Estévez et al., 2016b). Our study is the first to describe the phenotype of a Nervous System-IGF-I conditional knockout and demonstrate that brain IGF-I regulates adult hippocampal neurogenesis. We have published an article in the journal Frontiers in Bioengineering and Biotechnology reporting the biocompatibility of graphene with neural stem cells (NSCs), neurons, astrocytes and oligodendrocytes (Defterali et al., 2016). Our results support the use of graphene to study functional neuronal networks. We have studied the role of the Tbr1 transcription factor (Méndez-Gómez et al., 2011; Díaz-Guerra et al., 2013) during neurogenesis, gliogenesis, and neuronal migration in the olfactory bulb and cerebral cortex. With this aim, we have used gain- and loss-of function strategies of this factor in vivo and in cell culture. We are analyzing potential molecular targets of Tbr1. Keywords Alzheimer s disease, Parkinson s disease, neurogenesis, ips cells, NSCs, transcription factors, growth factors, graphene Publications Nieto-Estevez V., Oueslati-Morales C.O., Li L., Pickel J., Morales A.V., Vicario-Abejon C. Brain Insulin-Like Growth Factor-I Directs the Transition from Stem Cells to Mature Neurons During Postnatal/Adult Hippocampal Neurogenesis. Stem Cells. 2016;34(8): Nieto-Estévez V, Defterali Ç, Vicario-Abejón C. IGF-I: A Key Growth Factor that Regulates Neurogenesis and Synaptogenesis from Embryonic to Adult Stages of the Brain.Frontiers in neuroscience. 2016; 10. Defteralı Ç, Verdejo R, Majeed S, Boschetti-de-Fierro A, Méndez-Gómez HR, Díaz-Guerra E et al. In Vitro Evaluation of Biocompatibility of Uncoated Thermally Reduced Graphene and Carbon Nanotube-Loaded PVDF Membranes with Adult Neural Stem Cell-Derived Neurons and Glia. Frontiers in bioengineering and biotechnology. 2016;4. Research projects Code: PI2015-2/02. Title: Potencial patológico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal Investigator: Joan X. Comella Carnice. CIBERNED s collaboration: Yes. CIBERNED groups: G413, G415, G204, G108, G411. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 216

217 Program 2 Group: Carlos Vicario Abejón Código: Title: Alzheimer s disease apolipoprotein pathology for treatment elucidation and development. Principal Investigator: Carlos Vicario Abejón. CIBERNED groups: G108. Type: European. Funding agency: Comisión Europea. Funding: ,12. Duration: Code: SAF R. Title: Formación de neuronas en el ser humano y ratón adultos por activación de la reprogramación celular y la neurogénesis endógena. Principal Investigator: Carlos Vicario Abejón. CIBERNED groups: G108. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: SAF R. Title: Regulación de la multipotencialidad de células madre neurales humanas y la neurogénesis en modelos de alzhéimer esporádico y alzhéimer familiar. Principal Investigator: Carlos Vicario Abejón. CIBERNED groups: G108. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: CIBERNED Annual Report / 217

218 GROUP Miquel Vila Bover 109 Principal Investigator Miquel Vila Bover ICREA research professor Research team Celine Perier Ramón y Cajal researcher Jordi Bové Badell Miguel Servet researcher Marta Martínez Vicente Miguel Servet researcher Oriol de Fàbregues-Boixar Nebot Nebot, clinician Iria Carballo Carbajal Postdoctoral fellow, CIBERNED Ariadna Laguna Tuset Postdoctoral fellow, Beatriu de Pinós Sandra Franco Iborra Postdoctoral fellow Albert Torra Talavera Postdoctoral fellow Jordi Galiano Landeira Postdoctoral fellow Annabelle Parent Technician Thais Cuadros Arasa Technician CIBERNED Beatriz Rodríguez Galván Technician Jordi Romero Giménez Technician Neurodegenerative Diseases Research Group Vall d Hebron Research Institute Mediterranean Building, First Floor, Lab. 102 Pg. Vall d Hebron, Barcelona (Spain) Tel.: / Fax: mvila@ir.vhebron.net CIBERNED Annual Report / 218

219 Summary In 2016 we have continued our studies on the pathogenicity of α-synuclein and its potential role on neuron dysfunction/death in the context Parkinson s disease. We have also continued with the development of novel therapeutic tools to interfere with α-synuclein-mediated neurotoxicity. In addition, we have particiapted in a study in which we have shown that transplantation of hematopoietic stem and progenitor cells (HSPCs) in the substantia nigra of different mouse models of Parkinson s disease significantly ameliorates dopaminergic neuron loss and motor function. Transplanted HSPCs can fuse with neurons and with glial cells in the ventral midbrain of Parkinson s disease mice. The hybrids can undergo reprogramming in vivo and long-term survival after transplantation, while acquiring features ofmature astroglia. These newly generated astroglia were essential for functional rescue of the dopaminergic neurons. Our data indicates that glial-derived hybrids produced upon fusion of transplanted HSPCs in the substantia nigra can rescue the Parkinson s disease phenotype via a nichemediated effect and can be potentially exploited as an efficient cell-therapy approach for this disease. Keywords Parkinson s disease, α-synuclein, hematopoietic stem and progenitor cells Publications Garcia-Lezana T., Oria M., Romero-Gimenez J., Bove J., Vila M., Genesca J. et al. Cerebellar neurodegeneration in a new rat model of episodic hepatic encephalopathy. Journal of Cerebral Blood Flow and Metabolism. 2017;37(3): Epub García-Prat L, Muñoz-Cánoves P, Martinez-Vicente M. Dysfunctional autophagy is a driver of muscle stem cell functional decline with aging. Autophagy Garcia-Prat L., Martinez-Vicente M., Munoz-Canoves P. Autophagy: A decisive process for stemness. Oncotarget. 2016;7(11): Altarche-Xifro W., di Vicino U., Munoz-Martin M.I., Bortolozzi A., Bove J., Vila M. et al. Functional Rescue of Dopaminergic Neuron Loss in Parkinson s Disease Mice After Transplantation of Hematopoietic Stem and Progenitor Cells. EBioMedicine. 2016;8: Garcia-Prat L., Martinez-Vicente M., Munoz-Canoves P. Methods for mitochondria and mitophagy flux analyses in stem cells of resting and regenerating skeletal muscle. Methods in Molecular Biology. 2016;1460: Recasens A., Perier C., Sue C.M. Role of micrornas in the regulation of α-synuclein expression: A systematic review. Frontiers in Molecular Neuroscience. 2016;9. Research projects Code: PI2016/06. Title: Identificación de vías fisiopatológicas y biomarcadores candidatos en la fase prediagnóstica de la enfermedad de Parkinson. Principal Investigator: Miquel Vila Bover. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G601, G207, G410. Type: Intramurales. Funding agency: CIBERNED. Funding: Duration: CIBERNED Annual Report / 219

220 Code: 392/U/2014. Title: Role of the cellular prion protein as cross-talk protein between alpha-syn/ LRRK2 and p-tau in sporadic and familiar Parkinson s disease. Principal Investigator: José Antonio del Río (Fundació Institut de Bioenginyeria de Catalunya). CIBERNED s collaboration: Yes. CIBERNED groups: G109, G114. Type: Private. Funding agency: Fundació la Marató de TV3. Funding: Duration: Code: 2014\FTPGB-LAGUNA. Title: Estudio del perfil transcripcional en subgrupos de pacientes prodrómicos con enfermedad de Parkinson. Principal Investigator: Ariadna Laguna Tuset. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G207. Type: Private. Funding agency: Fundación Tatiana Pérez de Guzmán el Bueno. Funding: Duration: Code: SAF R. Title: Disfunción mitocondrial en la enfermedad de Parkinson: papel de la dinámica mitocondrial. Principal Investigator: Celine Perier. CIBERNED groups: G109. Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: PI13/ Title: Rol de la neuromelanina y alfa-sinucleina en el inicio y extensión de la enfermedad de Parkinson: potencial diagnóstico y terapéutico en nuevos modelos pre-clínicos. Principal Investigator: Miquel Vila Bover. CIBERNED groups: G109. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: Code: PI14/ Title: Rol patogénico de la disfunción autofágica/lisosomal en la enfermedad de Parkinson: diagnóstico y nuevas terapias pre-clínicas. Principal Investigator: Marta Martínez Vicente. CIBERNED groups: G109. Other CIBER s collaboration: Yes (CIBER-BBN). Type: International. Funding agency: Fondo de Investigación Sanitaria. Funding: Duration: Code: MJFF_TARGET.ADV-2015 (Grant ID: 11580). Title: Targeting the transcriptional regulator of autophagy LMX1B for Parkinson s disease therapeutic development. Principal Investigator: Ariadna Laguna Tuset. CIBERNED groups: G109. Type: International. Funding agency: The Michael J. Fox Foundation. Funding: Duration: Code: PI15/ Title: Respuesta inmune adaptativa en la enfermedad de Parkinson inducida por proteoformas de alfa-sinucleina presentes en los cuerpos de Lewy: aplicación diagnóstica y terapéutica. Principal Investigator: Jordi Bove Badell. CIBERNED groups: G109. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: Duration: 2016 Code: MJFF Staff Initiated Title: Prion-like dissemination of synuclein pathology: a non-human primate study. Principal Investigator: Erwan Bezard (Universite de Bordeaux, France). CIBERNED s collaboration: Yes. CIBERNED groups: G109, G205. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: Code: 288/C/2014. Title: Catalan Network of Multiple System Atrophy: Biomarkers and Pathophysiology. Principal Investigator: M.j. Marti (Hospital Clinic-IDIBAPS). CIBERNED s collaboration: Yes. CIBERNED groups: G109, G205. Type: Private. Funding agency: Fundació La Marató de TV3. Funding: Duration: CIBERNED Annual Report / 220

221 Program 2 Group: Miquel Vila Bover Code: 2014 SGR Title: Apoptosi i Neurodegeneracio (GRC). Principal Investigator: Joan Comella Carnicé. CIBERNED s collaboration: Yes. CIBERNED groups: G109, G413. Type: International. Funding agency: AGAUR. Funding: Duration: Code: Fundació la Marató de TV3-Malalties del cor (320/U/2015). Title: New molecular functions of apoptotic genes in cardiac development and stress. Principal Investigator: Daniel Sanchis Morales (IRBLLEIDA, Universitat de Lleida). CIBERNED groups: G109. Type: Private. Funding agency: Fundació la Marató de TV3. Funding: Duration: Code: RTC Title: Nuevas estrategias terapéuticas para el tratamiento de sinocleunopatias. Principal Investigation: Raquel Revilla (nlife Therapeutics). CIBERNED groups: G109. Type: National. Funding agency: Ministerio de Economía, Industria 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). CIBERNED groups: G109. Type: International. Funding agency: Michael J. Fox Foundation. Funding: Duration: Code: MJFF TPP2014. Title: A precise approach for nucleosidebased therapy of neuromuscular disorders with defects in mitochondrial dna. Principal Investigator: Miguel Ángel Martín Casanueva-Fundación Hospital 12 de Octubre. CIBERNED groups: G109. Other CIBER s collaboration: Yes (CIBERER). Type: National. Funding agency: Ministerio de Economía, Industria y Competitividad. Funding: Duration: Code: MJFF_TARGET ADV2016. Title: Targeting a novel transcriptional pathway to reduce alpha-synuclein expression. Principal Investigator: Miquel Vila Bover/ Solange Desagher (CNRS-Institute of Molecular Genetics Montpellier, France). CIBERNED groups: G109. Type: International. Funding agency: The Michael J. Fox Foundation. Funding: Duration: Code: COST Action CA Title: TRANSAUTOPHAGY-European Network of Multidisciplinary Research and Translation of Autophagy knowledge. Principal Investigator: Caty Casas (UaB). CIBERNED groups: G109. Type: European. Funding agency: Comisión Europea. Funding: N.D. Duration: 2016 PhD dissertations Author: Oriol de Fàbregues-Boixar Nebot. Title: Nuevas formas farmacéuticas y vías de administración de levodopa para el tratamiento de la enfermedad de Parkinson: nueve años de experiencia en infusión intraduodenal de gel de levodopa/ carbidopa. Date: 8 February Supervisor: Miquel Vila Bover CIBERNED Annual Report / 221

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223 Cooperative research

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225 Cooperative research The current cooperative research program continues to be viewed as an essential scientific tool for planning CIBERNED future actions. This program is one of the cornerstones of the Center 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 benefit of the research activity. The National Evaluation Agency (ANEP), although subject to final approval by the Steering Committee, externally reviews the proposals, so that the allocation of funds is independent and transparent. The cooperative research program has so far launched seven calls for projects in total. The first four (2010, 2011, 2013 and 2014) has already developed fifteen completed projects and a total accumulated budget allocation of These projects from the first four calls are listed below: 2010 call Code PI2010/05 PI2010/06 PI2010/07 PI2010/08 PI2010/09 PI2010/11 Title Generating a dopaminergic neuronal model from induced pluripotent stem cells from patients with Parkinson s disease associated to mutations in the LRRK2 gene Neuroprotection in Huntington s disease Reelin and GSK3 as therapeutic targets in Alzheimer s disease Glial activation during the neuroinflammatory process: a potential therapeutic target for Alzheimer s disease BESAD-P: Biomarkers of early stages of Alzheimer disease - prevention Consortium to generate a common database aimed at implementing clinical and basic research on neuromuscular diseases Código Coordinator Tolosa Sarró, Eduardo García de Yébenes Prous, Justo Ávila de Grado, Jesús Vitorica Fernández, Francisco Javier Ferrer Abizanda, Isidro Illa Sendra, Isabel 2011 call Code Title Coordinator PI2011/01 PI2011/02 PI2011/03 PI2011/04 The Nrf2 transcription factor as a new therapeutic target for Parkinson s disease Onset and progression of Parkinson s disease. Vulnerability of the nigrostriatal pathway, events at origin and destination Generation of dopaminergic neurons from somatic cells of parkinsonian patients with cognitive failure Multicenter study on LCR and neuroimaging biomarkers in the continuum preclinical- prodromal Alzheimer s disease (SIGNAL Study) Cuadrado Pastor, Antonio Obeso Inchausti, José Ángel Moratalla Villalba, Rosario Lleó Bisa, Alberto 2016 CIBERNED Annual Report / 225

226 2013 call Code PI2013/01 PI2013/05 PI2013/07 PI2013/08 PI2013/09 Title Emergent properties of the neuron-glia relationship underlying neurodegeneration and dementia in Alzheimer s disease Identification and molecular characterization of cannabinoid receptors subpopulations in poliglutaminopatías Role of GSK-3β in the cortical circuits alterations occurring in Alzheimer s disease Mitochondrial dynamics and mitophagy as therapeutic targets in Parkinson s and Huntington s diseases Synaptic degeneration and dysregulation of adult neurogenesis in murine models of neurodegeneration Código Coordinator Torres Alemán, Ignacio Guzmán Pastor, Manuel Iglesias Vacas, Teresa Soriano García, Eduardo Fernández Chacón, Rafael 2014 call Code PI2014/02 PI2014/06 Title Epigenetic mechanisms involved in the etiology and progression of rapidly progressive neurodegenerative dementias Onset and progression of Parkinson s disease: role of glial activation Código Coordinator Calero Lara, Miguel Rodríguez Oroz, Mª Cruz Those seventeen already finished projects have actively involved 57 CIBERNED research groups overall, in addition to some other external ones, representing 90,4 % of the existing groups. The 2014 call for CIBERNED cooperative projects ended in December As stated in the call remit, the groups involved addressed the obtained results in final reports that were provided to CIBERNED s scientific management. Below is a brief description of the results obtained by each of the projects from this 2014 call: Project PI2014/02: 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 A. Sánchez Juan, Pascual Institution CIBERNED, Carlos III Institute of Health - CIEN Foundation, Madrid CIBERNED, Bellvitge Biomecial research Institute, Barcelona CIBERNED, Catalonian Bioengineering Institute, Barcelona CIBERNED, Marqués de Valdecilla Foundation, Santander This project is focused on the study of epigenetic mechanisms involved in the etiology and progression of rapid progressive neurodegenerative dementias (rpndd) compared to their slow progressive counterparts. The main objectives are: 2016 CIBERNED Annual Report / 226

227 1) 2) Identification of epigenetic mechanisms that modulate the molecular expression of rpndd (CJD, rpad, and rplbd) in brain tissue. Analysis of the hypermutability of CpG dinucleotides in genes associated with CJD, AD and LBD as a regulatory mechanism of the expression of neurodegenerative dementias. 3) Targeted identification of epigenetic and transcriptomic signatures of differentiated IPSc. 4) 5) Device a sub-classification and definition of rpndd endophenotypes, as a function of neuropathological and molecular characteristics based on epigenetic signatures. Elucidate whether the observed changes can be used as biomarkers in accessible fluids such as blood and CSF. This work is a multidisciplinary approach common to different clinical entities with similar presentation, focused on the epigenetic mechanisms involve in the etiology of rpndd. Currently, the project is analyzing human samples from the Brain Bank of Institute of Neuropathology (Biobank HUB-ICO-IDIBELL, differentiated IPS cells, as well as ADN samples and blood and CSF samples from collections of the participants from several neurodegenerative disorders (CJD, rpad, rplbd, typical AD and LBD and age-matched controls). Following, we describe the most important results, arranged according to the pertaining objectives: Objective 1. Identification of epigenetic mechanisms that modulate the molecular expression of rpndd (CJD, rpad, and rplbd) in brain tissue. Results on this objective, led by Dr. Isidro Ferrer, indicated that rapid progressive dementias related to neurodegenerative disorders associated with abnormally-conformed protein deposits show alteration in: i) energy and mitochondrial metabolism, ii) purine metabolism, iii) inflammatory response, and iv) molecules involved in protein synthesis. As a result, eight papers have been published: five focused on prion diseases, and three on Parkinson s disease and Dementia with Lewy bodies. Another paper focused on rapid Alzheimer s disease has been submitted for publication. Objective 2. Analysis of the hypermutability of CpG dinucleotides in genes associated with CJD, AD and LBD as a regulatory mechanism of the expression of neurodegenerative dementias. The results of this objective, led by M. Calero, indicates: i) that the hypermutability of CpG dinucleotides may be involved in creating hot spots in the sequence of genes involved in neurodegenerative dementias, and ii) that variability in genes involved into the regulation of this hypermutability of CpG (DNA methyltransferase, cytosine deaminases, repair-enzymes, etc.) may be associated with neurodegenerative disorders. In this objective, we performed in silico sequence analysis of genes directly involved in neurodegenerative processes (PRNP, APP, PSEN1, PSEN2, SNCA), through the analysis of potential mutagenic sites and the study of the role of these changes in the functionality of the different proteins. We have also been able to determine several polymorphisms that appear to be associated with neurodegeneration (CJD and AD). Among them, 14 polymorphisms have been identified in the genes APOBEC1, DNMT1, DNMT3B, MBD4A and TDG that are associated with changes in the amino acid sequence in their corresponding proteins. Objective 3. Targeted identification of epigenetic and transcriptomic signatures of differentiated IPSc. During this year, in this objective led by Dr. Jose Antonio del Rio, several key tools have been generated: i) Spherical neural masses (SNMs) have been differentiated from neural induced pluripotent stem cell (IPSc) from a patient with GSS, and ii) The SNMs generated have been characterized and material for subsequent analyses (RNA, DNA, fixed cell cultures) prepared CIBERNED Annual Report / 227

228 Objective 4. Device a sub-classification and definition of rpndd endophenotypes as a function of neuropathological and molecular characteristics based on epigenetic signatures. Results of this objective will permit to delineate certain commonalities and differences among rapid dementias that will be comprehensively described after compilation of available data at the end of the project. Importantly, certain factors influence the rapidity of progression of Alzheimer s disease and Lewy body diseases, which is related on the one hand to the characteristics of soluble β-amyloid species, and, on the other, to the increased expression of certain pro-inflammatory cytokines and mediators of the inflammatory response. Objective 5. Elucidate whether the observed changes can be used as biomarkers in accessible fluids such as blood and CSF. The results of this objective, led by Dr. Pascual Sanchez Juan, following a comparative study between EA cases and controls has allowed us to select several mirna as candidate biomarkers, namely mir-9-5p, mir-134 and mir-598. Analysis of the routes involved by using the web page has demonstrated that these mirna are involved in: i) amyloid formation routes, ii) cellular stress signaling, and iii) Huntington disease. Additionally, Dr. Ferrer s team has studied the expression of malate dehydrogenase 1 as a possible CSF biomarker of sporadic Creuzfeldt-Jakob (scjd), and performed a literature review covering up-date knowledge on CSF biomarkers of neurodegenerative and vascular dementias. These work has been published in two papers. Other observations have been submitted for publication or are in progress as the utility of α-synuclein levels in combination with other markers in the diagnosis of prion diseases, and the expression levels of YKL-40 in dementias other tan Alzheimer s disease. The overall results obtained during the first year of the project are satisfactory, relevant and according to the initial work plan. It is important to remark that this collaborative project could not be carried out by any of the participants single-handedly; as the results are complementary and the project takes advantage and potentiates the technical and clinical expertise as well as the methodological resources of the research team, constituted. Project PI2014/06: Onset and progression of Parkinson s disease: role of glial activation Principal Investigator Rodríguez Oroz, Mª Cruz Matute Almau, Carlos Obeso Inchausti, José A. Rodríguez Díaz, Manuel Institution CIBERNED, Biodonostia Research Institute, San Sebastián CIBERNED, University of the Basque Country, Bilbao CIBERNED, Center for Applied Medical Research, Univ. Navarra, Pamplona CIBERNED, University La Laguna, Tenerife Background: Neuroinflammation and glial activation seems relevant in the aetiology and physiopathology of Parkinson s disease (PD). There is a strong interaction between inflammation and dopaminergic death, although neither human studies nor animal models used until this moment allow explaining the role of microglia and astrocytes in the initiation/progression of dopaminergic death in PD. In addition, it is not known whether LRRK2 mutations causing PD induce proinflammatory changes that can drive to dopaminergic degeneration. Objective: To study the role of astrocytes and microglia in the beginning and progression of dopaminergic death in PD CIBERNED Annual Report / 228

229 Design and Scope of the study: Traslational and multicenter study with complementary approaches using astrocycit cell cultures differentiated from induced pluripotent stem cells (ips) derived from patients, graft of these atrocytes in an animal model of selective dopaminergic lesion, and in vivo and postmorterm studies in two animal models of progressive parkinsonism. The study has been undertaken in 4 centers with in parallel and sequential complementary works comprising the clinic field (selection of patients for skin biopsies) and the basic field with two lines of research: 1. Isolation and differentiation of fibroblast from skin biopsies to ips and subsequently to astrocytes, graft of these astrocytes in the striatum of a model of selective dopaminergic lesion; 2. Development and sequential in vivo and post-mortem study of models of progressive parkinsonism in rat and monkey. Instrumentation: 1) 2) 3) Analysis of the relationship between glial cells, pro-inflammatory cytokines and death of dopaminergic neurons in in vivo (positron emission tomography (PET)) and post-mortem studies in the substantia nigra compacta (SNc) and striatum in two animal models of progressive nigro-striatal lesion (rat overexpressing mutant human a-syn A53T in the SNc and monkey with progressive MPTP intoxication) at different times corresponding to progressive higher degrees of dopaminergic lesion. Cell cultures and characterization of the inflammatory profile of astrocytes (Ast) differentiated from induced pluripotent stem cells (ips) derived from fibroblasts of sporadic and LRRK2 mutations PD patients and from asymptomatic carriers of this mutation. Graft of these astrocytes in a rat model of selective dopaminergic lesion and evaluation of the local inflammatory response and survival of dopaminergic neurons. Results: In the rat model of progressive parkinsonism it has been observed: a progressive dopaminergic depletion (30% in SNc at 3 weeks post-inoculation of the viral vector with α-syn); an early reduction of the microglial activation in the SNc (24h) antedating the dopaminergic death with a later increase at 3 weeks, suggesting that the microglial activation might be a consequence of the dopaminergic death; a lack of astrocytic activation at 24 h and a reduction at 3 weeks. The PET study shows an increment of microglial activity in the SNc respect to the striatum although no differences either between the times of study or between groups (dopaminergic lesion and control) have been observed. In the MPTP monkey model, the in vivo study with PET shows an increment in the striatal microglial activity at day 15 after the administration of the toxic, that is further augmented at day 45 only in animals with parkinsonism (not seen in asymptomatic monkeys with less severe dopaminergic lesion). On the other hand, a rat model of selective dopaminergic lesion without unspecific tissue damage has been developed showing that striatal astrocytes and microglia reactivity is different (early astrocytosis without microglial reactivity). The role (protection versus promotion of dopaminergic death) of reactive astrocytes has been studied showing that astrocytes have a key role in the autophagy of dopaminergic detritus without evidence of microglial participation in this process. These data are in keeping with the results obtained in the PET (in vivo) and postmortem studies in the model of progressive parkinsonism in rat where there is scarce microglial participation in early stages of the dopaminergic lesion (beginning of microglial activation at 3 weeks when dopaminergic lesion is 30%), and in the PETs of the monkey model, where the highest microglial activation is associated with a more severe dopaminergic lesion and parkinsonism. In addition, the characterization of the astrocytosis observed in the rat model of selective dopaminergic lesion shows some common features with the one associated with unspecific lesions (increment of GFAP, glutamine sinthetasa, S100β, NDRG2 and vimentine) but some differences (there is neither 2016 CIBERNED Annual Report / 229

230 astrocytic proliferation nor astrocytic differentiation from NG2 cells or collateral microgliosis). This a critical contribution to be able to evaluate the inflammatory changes specifically associated to the grafts of astrocytes derived from ips of patients. Simultaneously, a method for the striatal graft of astrocytes derived from 1-3 post-natal rats and obtained by selective culture or by magnetic nano-spheres has been developed. These studies provide optimal conditions to procedure with the grafts of astrocytes derived from patients. On the other hand, we have isolated fibroblast from skin biopsies of PD patients with and without LRRK2 mutation, asymptomatic carriers of this mutation and control subjects sex and age matched of which ips have been derived. Currently, we are differentiating astrocytes from ips to subsequently undertake their graft to the selective dopaminergic lesion rat model. Meanwhile, commercial astrocytes derived from ips of healthy controls have been bought in order to start this part of the study. Next month, once cultures of astrocytes have been obtained from the 4 groups of humans, the expression (q-pcr) and release (ELISA) of the main pro-inflammatory cytokines will be study to characterize the inflammatory profile of these astrocytes. Conclusion: The in vivo (PET studies) and postmortem results obtained in the animal models of progressive parkinsonism and selective dopaminergic lesion support that inflammation has an important role in the beginning and progression of PD and that astrocytes and microglia have different roles at different time of the evolution of the dopaminergic lesion. Astrocytes might decrease the rate of progression of dopaminergic degeneration in the early stages of the disease whereas microglia seems to be activated later on when the dopaminergic lesion is well established and parkinsonian signs are present. Our data also suggest that PET studies might be useful as in vivo maker of the microglial activity but it is necessary a correlation study that will be performed when all data have been quantified. ips have been derived from non-genetic and genetic (LRRK2) PD patients, asymptomatic carriers of the mutation and healthy controls and astrocytes will be obtained soon. Then, their pro-inflammatory characterization and the effect of their graft in the survival of dopaminergic cells in a rat model well characterized to this aim will be undertaken. In June 2015 the fifth call for CIBERNED cooperative projects was published, which was evaluated again by ANEP and resolved in early September with the award of three new collaborative projects, which are detailed below I call Code PI2015/01 PI2015/02 PI2015/03 Title Neuregulin gene therapy aimed at the treatment of motor neuron degeneration in ALS Validation of new therapeutic targets and biomarkers in Parkinson s disease Differential metabolic profiles in Parkinson s disease Código Coordinator Navarro Acebes, Xavier Labandeira García, José Luis Fuentes Rodríguez, José Manuel This 2015-I call for projects it is endowed with 345,000 annually for two years, of 690,000 overall, for the 11 CIBERNED research groups and 3 external associated groups participatin in all three projects. Below is a brief description of the scientific activity carried out by each of the projects during its first year of implementation: 2016 CIBERNED Annual Report / 230

231 Project PI2015/01: Neuregulin gene therapy aimed at the treatment of motor neuron degeneration in ALS Principal Investigator Navarro Acebes, Xavier Rodríguez Álvarez, José García Verdugo, José Manuel Sáez Valero, Javier Institution CIBERNED, Autonomous University of Barcelona CIBERNED, Autonomous University of Barcelona CIBERNED, Cavanilles Institute, University of Valencia CIBERNED, Miguel Hernández University, Elche Recent genetic and cellular evidences suggest an association of the neurotrophic factor Neuregulin 1 (Nrg1) and its receptor ErbB4 in Charcot-Marie-Tooth and Amyotrophic Lateral Sclerosis (ALS) among other neurodegenerative diseases. Thus, loss-of-function mutations in the Nrg1 receptor ErbB4 are associated with late-onset autosomal-dominant ALS. Nrg1 expression is reduced both in ALS patients and SOD1G93A transgenic mice. In addition, active Nrg1 requires sequential processing by α-secretase (ADAM proteins) or β-secretase (BACE1) and presenilin (PS)/ γ-secretase, proteases also involved in the processing of APP in Alzheimer s disease (AD). However, the pathogenic role of Nrg1 and its processing in motoneuron survival is not well understood. Our hypothesis is that Nrg1/ErbB4 signaling could be a relevant molecular pathogenic mechanism underlying neurodegeneration. Therefore, targeting Nrg1/ErbB signaling may represent a promising novel therapeutic approach for ALS and other neurodegenerative diseases. To investigate the role of Nrg1/ErbB4 system on motoneuron dysfunction and loss in ALS pathogenesis, we propose to examine Nrg1/ErbB4 signaling and its therapeutic targeting in a faithful mutant mouse model of ALS, harboring a point mutation in the Sod1 gene, which leads to motoneuron degeneration and a phenotype mimicking the usual clinic of sporadic and familial ALS. We have first characterized the expression of Nrg1 isoforms and ErbB receptors in spinal cord and skeletal muscles of SOD1G93A mice and ALS patients. We found a significant reduction of ErbB4 levels in muscles of both ALS mice and human patients. In the spinal cord of the ALS mice, Nrg1 type I levels increased along the progression of the disease, whereas Nrg1 type III levels were reduced; we have also observed progressively reduced levels of ErbB3 and ErbB4 receptors in the spinal cord of ALS mice with age. Previous studies have shown that processing of Nrg1 and its receptor ErbB4 by γ - or β-secretases and presenilin (PS)/β-secretase regulates Nrg1/ErbB4/3 signaling. Our results indicate a PS/ β-secretasedependent processing of Nrg1 and ErbB4 in the cortex of adult mice and an impairment of Nrg1 and ErbB4 processing in PS cdko mice during neurodegeneration. In addition, we have characterized a soluble ectodomain fragment of ErbB4 (NTF) in human CSF. Binding of Nrg1 to ErbB4 leads to its secretase-cleavage and release of the kda ErB4 ectodomain fragment. Light and electron microscopy evidenced clear signs of cell damage in the somas of motoneurons of SOD1G93A mice, including overall reduction in cell volume and reduction of the number and length of active synaptic contacts. The influence of Nrg1 overexpression on these abnormalities will be analyzed. We have assayed the overexpression of Nrg1-I in the skeletal muscle of SOD1G93A mice by means of an AAV1 vector, and demonstrated that Nrg1-I promoted collateral reinnervation of denervated endplates, opening a new window for developing novel ALS therapies for functional recovery rather than preservation. Consequently, a more efficient approach is being pursued by intravenous administration of an AAV8 coding for Nrg1 type I under the regulation of the human desmin promoter, specific for muscle cells that will allow us to target a large number of muscles. Altogether, the results obtained demonstrate that Nrg1 is involved in the two main processes of ALS, motoneuron degeneration and loss of neuromuscular contacts. The gene therapy assay increasing the expression of Nrg1 in muscles of the ASL mice showed a striking improvement of the innervation of injected muscles CIBERNED Annual Report / 231

232 Project PI2015/02: Validation of new therapeutic targets and biomarkers in Parkinson s disease Principal Investigator Labandeira García, José Luis Cuadrado Pastor, Antonio Lanciego Pérez, José Luis Kulisevsky Bojarski, Jaime Institution CIBERNED, University of Santiago de Compostela CIBERNED, Autonomous University of Madrid CIBERNED, Center of Applied Medical Research, Univ. Navarra, Pamplona CIBERNED, Santa Creu i Sant Pau Hospital, Barcelona The development of new therapeutical strategies against neurodegenerative diseases such as Parkinson s disease (PD) is a major objective in biomedical research. New strategies going beyond current symptomatic treatments are necessary. Unfortunately, all neuroprotective strategies have failed in clinical trials. The lack of therapies against cognitive impairtment in PD is particularly problematic. The present project addresses these problems through the following objectives: 1) 2) 3) 4) A new animal model of PD induced by alfa-synuclein introduced with viral vectors will be developed and optimized. We will address two new therapeutical targets related to major mechanism of the redox metabolism and neuroinflammation: inhibition of the proinflammatory axis angiotensin/nadphoxidase/rho/kinase, and activation of the transcription factor Nfr2, a master regulator of the anti-oxidant defense system. Alterations observed in rodent and primate preclinical models will be correlated with observations in postmortem brain tissue from PD patients. Advanced Glycation End products (AGEs) and other potential biomarkers markers will be determined in samples from plasma of PD patiens and controls to find possible biomarkers of cognitive impairtment in PD. The results on these biomarkers will be correlated with those observed in primate models of PD. Finally, the effects of the suggested therapies on the above mentioned biomarkers will be tested in rodent and primate animal models of PD. Project PI2015/03: Differential metabolic profiles in Parkinson s disease Principal Investigator Fuentes Rodríguez, José Manuel Pérez Castillo, Ana María Pérez Tur, Jordi Institution CIBERNED, University of Extremadura, Cáceres CIBERNED, Institute of Biomedical Research CSIC-UAM, Madrid CIBERNED, Institute of Biomedicine of Valencia, CSIC Intermediate metabolism and cell signaling pathways have been considered and studied to date as ndependent entities. But in reality catabolic and anabolic pathways are which provide cell energy and precursors required for cell survival. On the other hand subcellular organelles with important metabolic functions, such as mitochondria play a key role in the origin of a large number of diseases including Parkinson s disease. Therefore the knowledge of metabolic variations to integrate and understand the mechanisms leading to neurodegeneration is essential. The determination of differential metabolic profiles in models of Parkinson s disease may allow the development of a diagnostic pattern, risk and prognosis of it CIBERNED Annual Report / 232

233 Objectives: Objective 1: Identification of transcriptome and metabolomics differential profiles in: 1.1 Cell models overexpressing pathogenic G2019S and R1441G pathogenic forms of LRRK In murine models overexpressing G2019S and R1441G pathogenic forms of Lrrk In human fibroblasts from healthy individuals, idiopathic Parkinson patients and asymptomatic carriers and patients with G2019S and R1441G pathogenic forms of LRRK In blood plasma from the same donors described in 1.3. Objective 2: Identification of transcriptome profiles and differential models described in 1.1 metabolomics., 1.2. and 1.3. after exposure to the parkinsonian neurotoxin MPP + (cellular models) or MPTP (murine models). Objective 3: Validation of transcriptomic and metabolomic results by determining the variation of the altered proteins involved in metabolic processes (molecular identification of drug targets). Objective 4: Establishment of cellular and animal models of Parkinson s disease on which test the effect of modulators / metabolic adapters as potential neuroprotective agents for Parkinson s disease. This project is therefore the first combined transcriptomic and metabolomics study aimed at identifying specific genetic biomarkers and neurotoxic models and with a simultaneous study in cellular systems, animals and samples from patients with idiopathic Parkinson, patients Parkinson pathogenic mutations for LRRK2 (G2019S and R1441G) and healthy carriers of the same mutation. It is a continuation of work previously done by our research group in the characterization of the deregulation of autophagy in these same cell models from Parkinson s patients, as a metabolic stress can lead to increased basal levels of autophagy, we hope to observe important differences especially in relation to redox and energy production systems. In addition, during the second half of 2015 a new call for projects (2015-II ) was launched, which resulted in the award of two additional projects in early 2016: 2015-II call Code PI2015-2/02 PI2015-2/06 Title Pathological potential of astrocytes: a new perspective on Alzheimer s disease Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration Código Coordinator Comella Carnicé, Joan Xavier Muñoz Cánoves, Pura This 2015-II call for projects it is endowed with annually for two years, oadding up to overall for the ten CIBERNED research groups that participate in the projects. A summary of the objectives set for the proposals approved in this fourth call, initiated in early 2016 is presented below: 2016 CIBERNED Annual Report / 233

234 Project PI2015-2/02 Pathological potential of astrocytes: a new perspective on Alzheimer s disease Principal Investigator Comella Carnicé, Joan Xavier Vitorica Ferrández, Francisco Javier Gutiérrez Pérez, Antonia Vicario Abejón, Carlos Moratalla Villalba, Rosario Institution CIBERNED, Vall d Hebron University Hospital, Barcelona CIBERNED, University of Seville CIBERNED, University of Malaga CIBERNED, Cajal Institute CSIC, Madrid CIBERNED, Cajal Institute CSIC, Madrid The present project, which can be considered as an extension of the past collaborative project in which we characterized the emergent roles of the interaction neuron/glia, aims at characterizing astrocyte function in Alzheimer s disease (AD). Although astrocytes have well characterized roles related with trophic support, homeostasis, elimination of toxic molecules, production of cytokines, vascularization, extracellular matrix reshaping and control of synaptic activity, it is unknown their specific role in the development of AD. It has been characterized that astrocytes activate and accumulate around β-amyloid plaques, but it is unknown the detailed changes in their genetic expression patterns, or how their secreted factors contribute to the development of the disease. In this project we propose to study the intrinsic characteristics of astrocytes from AD patients, that is, which are their molecular differences in comparison with astrocytes from healthy individuals; in addition, we want to study how these activated astrocytes from patients could be affecting or modulating the activity of other cell types in the brain, particularly neurons. To achieve this goal, we will use different cell and animal models we have been generating in our laboratories in the past collaborative project, particularly ips cells derived from patients fibroblasts differentiated into astrocytes (or even neurons), which will harbor the genetic footprint of the donors, and also different transgenic models, such as the transgenic overexpressing FAIM-L in neurons, and the classical models of AD with which we can cross our model (APP/PS1, 3xTg, etc). In order to characterize the astrocyte activation per se (objective 1), we will use the astrocytes obtained from patients ips, which we developed in the past collaborative project. We will pay special attention in the ApoE genotype of astrocytes, and will characterize their expression patterns through arrays or RNA-seq after β-amyloid or tau activation. On the other hand, we will try to determine the cause of the astrocyte hyperactivation and its pathologic consequences on other cells (objective 2), by using different animal models where the glial response is altered (diminished or impaired), in order to mimic what happens in patients. Finally, an essential component of neurodegenerative diseases is neuronal loss. In the former collaborative project we demonstrated a correlation between the levels of a neuron-specific death receptor antagonist, FAIM-L, and the degree of the disease. Thus, FAIM-L appears as a likely therapeutic target to stop or delay neurodegeneration. However, it is not known if astrocyte activation can modulate FAIM-L levels, and thus if this activation could play a role in the loss of protection of FAIM-L against neuronal death CIBERNED Annual Report / 234

235 Project PI2015-2/06: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration Principal Investigator Muñoz Cánoves, Pura Fariñas Gómez, Isabel Lucas Lozano, José Javier Fernández Chacón, Rafael Iglesias Vacas, Teresa Institution CIBERNED, Pompeu Fabra University, ICREA, Barcelona CIBERNED, University of Valencia Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid CIBERNED, University of Seville CIBERNED, Institute of Biomedical Research CSIC-UAM, Madrid The functional decline of the nervous and the musculoskeletal systems is a major factor determining the quality of life in elderly populations. Nowadays, it is well established that stem cells in specific neural and muscular niches, active throughout life, are essential for the functional maintenance of the brain and skeletal muscles. Stem cell properties, though, are sensitive to aging and to pathogenic factors such as molecular insults underlying neurodegeneration. Nevertheless, the molecular mechanisms that progressively hinder stem cells regenerative potential are poorly understood. Our consortium will focus on the analysis of key pathogenic processes such as: (1) the agingdependent inhibition of autophagy in stem cells mediated by target of rapamycin (mtor) signaling pathway, (2) the premature exhaustion of stem cell pools due to cell cycle and cell division mode dysregulation and (3) stem cell dysfunctions due to alterations in alternative splicing such as those occurring in neurodegenerative diseases associated to polyglutamine repeats. The coordinator of the consortium Dr. Muñoz-Cánoves has recently described key molecular pathways underlying the functional decline and senescence of satellite cells that impair muscle rejuvenation (Sousa-Victor, 2014). Interestingly, a thorough unbiased transcriptomic analysis included in that study has identified the mtor signaling pathway as a potential culprit in stem cell aging and, remarkably, several other molecular targets that are major subjects of study in the laboratories of the other consortium s members. Some of these targets include: (a) Cysteine String Protein-alpha (CSP-alpha), a synaptic co-chaperone that prevents presynaptic degeneration (García-Junco et. al, 2010) that, unexpectedly, maintains the postnatal quiescence of neural stem cells through inhibition of mtor (unpublished work from Dr. Fernández-Chacón s group); (b) Kinase D interacting substrate (Kidins220), a major effector of multiple receptor signalling pathways regulating cellular responses in nervous, vascular and skeletal muscle systems (Iglesias, 2000; Neubrand, 2012) that is present in neural stem cells (unpublished data from collaboration between Dr. Iglesias and Dr. Fariñas groups), supporting a potential role of Kidins220 in regulating NSC function; (c) Rbfox1, a neuron and muscle specific splicing factor previously linked to muscle dysfunction in expanded CUG-caused myotonic dystrophy DM1, that is downregulated in Huntington s disease brain thus leading to a pathogenic alteration of CPEB4 splicing (unpublished work from Dr. Lucas group). Our consortium brings together groups with ample expertise in the biology of stem cells and neural development, the role of satellite cells in muscle fiber maintenance and the molecular basis of neurodegenerative diseases and synaptic dysfunction. Our laboratories will synergistically interact to maximize multidisciplinary methodological resources and expertise which include a highly specialized knowledge in the cell biology of neural stem and satellite cells, yet-unpublished biological resources, i.e. mouse models, and first-hand understanding of specific signaling molecules and pathways. We expect that our project will throw light on key mechanisms involved in the functional maintenance of adult stem cells, an essential point for the potential regeneration of muscle fibers and specific neuronal types CIBERNED Annual Report / 235

236 Finally, during the second half of 2016, the seventh call for projects was launched. This call was resolved at the end of 2016, resulting in five new funded projects call Code PI2016/01 PI2016/02 PI2016/04 PI2016/05 PI2016/06 Title Alterations of gluco-lipid metabolism and development of Alzheimer s dementia Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches The ALS CIBERNED Challenge: Accelerating New Drug Discovery Dream inhibitors and Alzheimer s Disease Identification of pathophysiological pathways and candidate biomarkers in the prediagnostic phase of Parkinson s disease Código Coordinator Torres Alemán, Ignacio Del Río Fernández, José Antonio López de Munain Arregui, Adolfo Naranjo Orovio, José Ramón Vila Bover, Miquel This 2016 call for projects it is endowed with 508,000 annually for two years, of 1,016,000 overall, for the nineteen CIBERNED research groups that participate in them. A summary of the objectives set for the projects awarded in the seventh call, initiated at the end of 2016 is presented below. Project PI2016/01: Alterations of gluco-lipid metabolism and development of Alzheimer s dementia Principal Investigator Torres Alemán, Ignacio Wandosell Jurado, Francisco Camins Espuny, Antonio Carro Díaz, Eva Cantero Lorente, José Luis Institution CIBERNED, Cajal Institute CSIC, Madrid CIBERNED, Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid CIBERNED, University of Barcelona CIBERNED, Doce de Octubre University Hospital, Madrid CIBERNED, Pablo de Olavide University, Sevilla In this project we propose a parallel study at both clinical and experimental levels. The idea is to transfer what is observed in humans to animal models with the idea of deepening our knowledge of the potential pathogenic mechanisms, more specifically those referring to imbalances in glyco-lipid metabolism. On the one hand, we propose an analysis of the lipopolysaccharide profile in a cohort of people that includes the whole spectrum of the natural history of dementia, from normal aging until Alzheimer s disease (AD) - going through the phases of subjective cognitive decline (SCD) and mild cognitive impairment (MCI). We will also carry out structural (volumetry and cortical thickness) as well as functional (glucose PET) brain imaging studies, with the aim of obtaining potential imaging biomarkers to which we can relate to the peripheral metabolism and could define each AD stages. The main peripheral biomarkers identified in this first group will be validated in a cohort composed of more than 2,000 individuals and we will try to transfer to the different animal models proposed in the present study CIBERNED Annual Report / 236

237 In animal models of AD, a battery of genetic and biochemical tools will be used to explore a possible metabolic basis of AD. More specifically, we want to examine how the metabolic imbalances associated with either an unbalanced diet (high in fat) or alterations in cerebral glucose metabolism (typical of AD), affect the development of AD. We will use the APP/ PS1 familial AD mouse model as the basic experimental paradigm in which we will eliminate: 1) the JNK1 kinase globally (previous studies indicate its relevance in insulin signaling), or 2) insulin or IGF-I receptors in astrocytes (which lead to profound changes in the cerebral glucose metabolism). In order to model the impact of an unbalanced diet in disease onset and progression, we will combine the cerebral alterations present in the APP/PS1 mouse model with the metabolic changes induced by exposure to a high fat diet. Using this general approach, we will examine the role of JNK1 (involved in the phosphorylation of insulin receptor substrates), the influence of the ovarian hormone estradiol (because of the higher incidence of AD in women), and the role of amyloidosis -for the latter study we will use the EMD mouse model (lacking megalin in cerebral endothelium) that develops EA symptoms in the absence of amyloidosis. We will measure the different parameters associated with AD (amyloidosis, tauopathy, cerebral hypoperfusion) and the metabolic syndrome (dyslipidemia, insulin resistance). In a fourth experimental group, determining the same parameters, we will analyze the influence of changes in cerebral glucose metabolism on the development of AD (mutant mice lacking IR or IGF-IR in astrocytes). In order to explore potential pathogenic mechanisms resulting from these central and peripheral changes, macroautophagy mechanisms related to lipid alterations and in situations of insulin resistance (generated by a fat-rich diet or lack of insulin receptors in astrocytes) will be analyzed in vitro and in vivo, aimed at determining the role of this process in the development of proteostasis dysfunction associated with AD. Project PI2016/02: Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches Principal Investigator Del Río Fernández, José Antonio Ávila de Grado, Jesús Canela Campos, Enric Isidre Institution CIBERNED, Catalonian Institute de Bioengineering, Barcelona CIBERNED, Center for Molecular Biology Severo Ochoa CSIC-UAM, Madrid CIBERNED, University of Barcelona Numerous aged-related neurodegenerative diseases (ND) are characterized by the slow but inexorable advance of neuronal dysfunctions in brain patients that progress though anatomically linked regions. This evolution of the disease shared with the progressive accumulation of abnormally folded proteins in affected neurons that stem for gradual neuronal dysfunction, which involves synaptic transmission abnormalities, axonal damage and cell death. However, in some cases the observed spread though brain parenchyma failed to follow the classical progression illustrated in seminar studies, e.g., in Alzheimer s (AD) or Parkinson s (PD) diseases. For example, some elderly individuals without diagnostics showed Lewy bodies in the olfactory bulb6, amygdala or cortex which should seem to violate the theory of progression and perhaps fit better with a multicentric disease process. In addition, anatomically connected regions of the brain (e.g., entorhinohippocampal formation) showed different p-tau burden in AD and other tauphaties. In conclusion, these data suggested both an area-specific and a differential neuronal vulnerability during disease progression. Factor/s modulating this differential neuronal vulnerability are unknown. ND and lab-on-chip devices. Our knowledge of many ND relies on the possibility of analysing neural samples only at symptomatic stages of the disease. Indeed, changes during asymptomatic stages of these diseases cannot be fully analysed, and, more relevantly, a prodromal study during 2016 CIBERNED Annual Report / 237

238 illness evolution cannot be developed properly. To overcome this, various experimental models are used to study brain syndromes. They range from whole animal models to dissociated cell culture systems. Very recently, advances in the field of cell culture, stem cell biology and lab on a chip technologies have further advanced the prospect of in vitro system to model some neurodegenerative diseases. Among emerging in vitro technologies using organoids or induced pluripotent stem cells in 3D cultures; microfluidic devices (MFD) are a powerful technique to monitor neurodegeneration and its associated processed. Notably, microfluidic devices allowing the physical separation of neuronal soma from axons, were used to study neuronal cell behavior and physiology, and recently protein spreading (α-synuclein; β-amyloid16 or Tau ). The goal of the present project is to recreate in vitro using MFDs normal and diseased neuronal networks of selected neural pathways susceptible to suffer neurodegeneration by using mice disease models or differentiated human pluripotent stem cells. We would like to focus on α-synuclein and Tau conformers. In particular, MFDs in our project will allows us for i) the recreation of specific neuronal networks of neuroanatomical relevance in neurodegenerative diseases in vitro; ii) to investigate factors mediating the propagation of the neurodegenerative process in models of ND and iii) to monitor early/late physiological changes mediated by amyloid proteins. Due that α-synuclein and Tau behave as prion-like elements, we will study in our culture platforms whether crucial neural functions (e.g., neuronal networks dynamics) or the onset and progression of neuronal dysfunctions appeared during the spreading of deleterious signals triggered by these prion-like elements in order to understand their propagation and neuron specific effects in vivo in parallel experiments. Project PI2016/04: The ALS CIBERNED Challenge: Accelerating New Drug Discovery Principal Investigator López de Munain Arregui, Adolfo Ferrer Abizanda, Isidro Fernández Ruiz, Javier Soriano García, Eduardo Institution CIBERNED, Biodonostia Research Institute, San Sebastian CIBERNED, Bellvitge Institute of Biomedical Research, Barcelona CIBERNED, Complutense University of Madrid CIBERNED, University of Barcelona Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive weakness and atrophy of skeletal muscles until paralysis and death. Multi-factorial and multisystemic neurotoxic mechanisms are the cause of a common pathological entity: motor neuron (MN) degeneration in central nervous system (CNS). The disease may spread to frontal cortex and other regions leading to frontotemporal lobar degeneration (FTLD) and dementia. The enormous heterogeneity in the patterns of clinical symptoms, anatomical onset, disease progression and survival among patients suggests that pathogenic pathways for MN degeneration are variable at topographical and pathophysiological levels. Moreover, involvement of the frontotemporal cortex in ALS and the motor system in FTLD-TDP-43 links ALS and FTLD-TDP-43 within the spectrum of TDP-43 proteinopathies. This complex scenario for ALS would explain the unfavorable outcomes collected in clinical trials. Future research must consider: 1) 2) 3) Identification of clinico-pathological markers in brain, CSF and blood specific of ALS subtypes, which will allow the design of individual intervention treatments. Identification of new altered molecular pathways in different CNS regions to unveil common and specific profiles of regional vulnerability. Identification of converging pathways of MN and frontal cortex degeneration, which will allow the generation of neuroprotective strategies to halt MN degeneration CIBERNED Annual Report / 238

239 The present collaborative project integrates 4 CIBERNED clinical/basic research groups (Adolfo López-de-Munain, Isidro Ferrer, Eduardo Soriano and Javier Fernández-Ruiz), with the purpose to address these 3 hot points in ALS/FTLD-TDP43 complex. Collaborative efforts will be extended to other CIBERNED groups. The first part of the project will create a unique Spanish ALS cohort by unifying several Spanish individual registries with standardization of clinical data and methods of biological sample collection. Large ALS cohort provides necessary potency to achieve the following milestones: 1) Identification of novel clinical and molecular signatures categorizing different ALS subtypes. 2) Identification of new altered molecular pathways in the motor system and frontal cortex in post-mortem brains of ALS (in comparison with FTLD-TDP-43). 3) 4) Discovery of novel biomarkers in brain, CSF, blood and striate muscle for diagnosis and prognosis. Drawing a pathophysiological map of the disease to detect potential targets for treatment. These objectives will be accomplished by carrying out combined omics and computational integrative analyses in samples from alive ALS patients and post-mortem tissues. The second part of the project is aimed at exploring neuroprotective targets following the hypothesis of a crosstalk between TDP-43 proteinopathy and mitochondrial defects as a common pathogenic pathway to MN degeneration (and probably frontal/temporal cortex degeneration. Three main points will be analyzed: 1) 2) 3) Efficacy of patented compounds as novel inhibitors of RyR Ca2+ channel as a way to protect neuromuscular system from cytosolic Ca2+ overload induced by pathological TDP-43-mediated mitochondria defects. Study of pharmacological targets within the endocannabinoid system against TDP-43-mediated mitochondria defects. The role of Eutherian Armcx proteins on mitochondrial dynamics in relation to TDP-43 pathology as a promising target for MN neuroprotection. Both parts of the project are interconnected and will be carried out by the collaborative work of the four group members of the proposal. Project PI2016/05: DREAM inhibitors and Alzheimer s disease Principal Investigator Naranjo Orovio, José Ramón De Felipe Oroquieta, Javier Ceña Callejo, Valentín Institution CIBERNED, National Center of Biotechnology, CSIC, Madrid CIBERNED, Cajal Institute CSIC; Technical University, Madrid CIBERNED, University of Castilla La Mancha, Albacete Deregulated protein and Ca2+ homeostasis underlie synaptic dysfunction and neuronal death in neurodegenerative diseases including Huntington s (HD) and Alzheimer s disease (AD). Early symptoms of synaptic dysfunction in AD are characterized by a reduced response to external stimuli and a progressive loss of post-synaptic dendritic spines in excitatory synapses, which correlates strongly with AD symptoms and memory loss. Previous to spine loss, atrophy of the spine apparatus, a synaptopodin-positive ER-associated membranous structure, is particularly relevant since the spine apparatus has an essential role in regulating the fate of calcium entering the spine upon synaptic activation and could be considered the structural base for synaptic plasticity CIBERNED Annual Report / 239

240 Dendritic failure in AD is also largely related to the accumulation of pathogenic protein aggregates that have been related to a defective unfolded protein response (UPR) and to changes in actin polymerization as a result of increased dephosphorylation of cofilin in AD. Recently, we have shown that repaglinide, an oral-hypoglycemic agent, enhanced ATF6 processing, which improved the pro-survival function of the unfolded protein response (UPR), and reduced neuronal loss in the striatum of R6/2 mice, a mouse model of HD. The mechanism involves the disruption by repaglinide of the calcium-dependent interaction between ATF6 and the neuronal calcium sensor DREAM, which results in the nuclear accumulation of transcriptionally active ATF6. Improved UPR after chronic repaglinide administration resulted in delayed onset and slowed progression of disease symptoms including loss of motor coordination and cognitive capabilities. DREAM, also known as calsenilin or KChIP3, is a Ca2+ binding protein that regulates Ca2+ homeostasis and neuronal survival through transcriptional control of target genes and through protein-protein interactions. DREAM was originally associated with AD because of its interaction with presenilins. Preliminary results from our group show changes in the expression of DREAM in cerebral cortex and hippocampus in J20 and Tg2576 mice, two murine models of AD, as well as in frontal cortex from AD patients. Genetic manipulation of DREAM levels or chronic repaglinide treatment modified cognition in Tg2576 and J20 mice. These data support the idea that, like in HD, an early down regulation of DREAM level in neurons during the pre-symptomatic phase of the AD, is part of a neuroprotective mechanism and suggest that DREAM could be a novel and wide spectrum target for therapeutic intervention in AD. The aim of this collaborative research program is to investigate whether cognition improvement in AD mouse models after repaglinide restores the early response to neuronal activation (induction of immediate-early genes Npas4, Nr4a1 and Arc) and is related to changes in synaptopodin-positive dendritic spines, in cofilin dephosphorylation and/or in increased ATF6 processing and UPR activation in hippocampal neurons. Project PI2016/06: Identification of pathophysiological pathways and candidate biomarkers in the prediagnostic phase of Parkinson s disease Principal Investigator Vila Bover, Miquel Tolosa Sarró, Eduardo Trullas Oliva, Ramón Infante Ceberio, Jon Institution CIBERNED, Vall d Hebron University Hospital, Barcelona CIBERNED, Hospital Clinic of Barcelona CIBERNED, Institute of Biomedical Research IDIBAPS-CSIC, Barcelona CIBERNED, Marqués de Valdecilla Research Institute, Santander Mutations in the Leucine-rich-repeat-kinase-2 (LRRK2) gene are the most common cause of genetic Parkinson s disease (PD). Like idiopathic PD (ipd) the onset of LRRK2-PD is thought to occur years before onset of motor symptoms but information on prodromal LRRK2-PD is sparse. The study of non-manifesting-lrrk2-mutation-carriers (LRRK2-NMC), which are at increased risk of developing LRRK2-PD, represents a unique opportunity to investigate the early molecular pathophysiology of LRRK2-PD and to identify molecular signatures that could be candidate biomarkers associated to prodromal and/or manifest LRRK2-PD. In this proposal we will undertake four different and complementary approaches in human biological samples selected to address these fundamental questions. In a first discovery phase, we will study LRRK2-NMC with normal and abnormal DAT-SPECT imaging characteristics, LRRK2- PD, ipd and healthy controls, from whom biological samples and exhaustive clinical data has already been collected. In these five groups we will study the serum metabolomic profiles and we will characterize the epigenome of the four major cell types represented in peripheral blood, to compare to previous differences found in ipd patients and to explore whether metabolomic and epigenomic changes occur at the prodromal phase of LRRK2-PD CIBERNED Annual Report / 240

241 In addition, we will assess cell-free mitochondrial DNA (mtdna) content and expression in the same serum and blood samples, to expand previous findings in cerebrospinal fluid (CSF) and to explore the relationship between mtdna copy number and metabolomic and epigenomic profiles. We will also study mtdna copy number and release in patient-derived fibroblasts to determine whether differences in serum, blood and CSF contents between groups are linked with altered mtdna replication or release. Finally, we will use an integrative systems biology approach to identify key pathophysiological pathways and molecular signatures associated to prodromal and/or manifest LRRK2-PD. In a second validation phase, the most relevant candidate biomarkers identified in the first discovery phase will be validated in additional samples from the five groups of interest in this study. We expect that our multidisciplinary collaborative effort will allow the identification CIBERNED cooperative program has so far invested an overall budget of about , distributed among the 27 cooperative projects mentioned above. Up to 57 CIBERNED research groups have actively participated in the cooperative program, which represents 90.4 % of all groups belonging to CIBERNED during this period, along with a dozen of outside associated groups. In addition, 49 groups (77,7 %) have participated in more than one cooperative project and projects 2015-I, 2015-II and 2016 are currently still active, with the involvement of 40 research groups CIBERNED (75,4%). By analyzing previous calls, and considering that some are already closed and others still ongoing, an estimation of the budget invested can be made by individual project and participating group. Thus: Call Budget Duration in years Number of groups Number of projects Average budget for project and year Average budget for project and group I 2015-II , , , , , , , , , , , , , , CIBERNED Annual Report / 241

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243 International relations

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245 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. This global concern has led to the creation of the World Dementia Council (WDC) with the aim of collectively spur action against dementia worldwide in the areas of research, clinical care and social awareness. 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. As a result, WHO has decided to establish a global observatory on dementia to monitor the prevalence of the condition and resources to care for patients in Member States as well as to track the establishment of national plans and policies against dementia. 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 CIBERNED Annual Report / 245

246 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, four transnational projects were awarded for the period Subsequently, in late 2012 a new call under the topic Identifying protective factors and genetic and epigenetic risk in neurodegenerative diseases was launched, resulting in five approved projects for the period The projects involving CIBERNED research groups that have been active during 2016 are described be 2016 CIBERNED Annual Report / 246

247 COURAGE-PD: Comprehensive unbiased risk factor assessment for genetics and environment in Parkinson s disease Overall budget Starting date Duration Coordinator CIBERNED participation 4,5M January, years Thomas Gasser, University of Tuebingen, Germany Eduardo Tolosa group Project partners: Thomas Gasser & Rejko Krüger University of Tuebingen, 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, Instituto de Medicina Molecular, Portugal Eduardo Tolosa, Universitat de Barcelona, Spain Rudi Balling, University of Luxembourg, Luxembur Peter Heutink, VU University Medical Centre, Amsterdam, Holland In 2016, COURAGE-PD has mainly be working on the following tasks of the work programme: Work package 1: Identification of genetic risk variants: Task 1: Exome sequencing. Task 2: Resequencing of GWAS loci. T ask 3: Bioinformatic analysis. Work package 2: Genetic susceptibility, protective factors, and gene-gene and gene-environment interactions in PD. Task 1: COURAGE-GEOPD chip development and analysis. Task 4: Improving causal inference for environmental factors in PD: MR, GEI, GGI analyses. Work package 3: Task 1 and Task 2: Functional validation of novel genes for monogenic PD and functional characterization of newly identified genetic risk or protective factors. Task 3: Modeling gene-environment interactions (GEI) in vitro. Task 4: Transcriptional analyses of mda neurons expressing novel genetic causes for PD. More specifically, the Tolosa group (CIBERNED) has been directly involved in the following activities: 2016 CIBERNED Annual Report / 247

248 First, all DNA samples from several selected Spanish families with several PD affected members have been already acquired and preliminary quality control on DNA has been performed. The workflow of the next-generation-sequencing project has been arranged, and genome sequencing has been performed in 2015, including other samples from different European countries. In addition, we provided around 500 samples (PD patients and controls) for the WP2, in order to be analysed with the COURAGE-GEOPD chip, which will cover all variants defined in the screening approach (WP1). Second, We recently published a genomewide DNA methylation and a transcriptome studies in induced pluripotent stem cell (ipsc)-derived dopaminergic neurons (DAn generated by cell reprogramming of somatic skin cells from patients with monogenic LRRK2-associated PD (L2PD) or sporadic PD (spd), and healthy subjects) (Fernández-Santiago et al 2015) by using the Infinium HumanMethylation450 BeadChip. We observed extensive DNA methylation changes in PD DAn, and of RNA expression, which were common in L2PD and spd. The PD-associated hypermethylation was prominent in gene regulatory regions such as enhancers and was related to the RNA and/or protein downregulation of a network of transcription factors. In the context of COURAGE project, we also have performed a whole genome bysulphite sequencing analysis of the DNA from these samples in order to have a complete knowledge of the epigenetic alterations in this model of PD. Actually, we are analysing this genomic data with two main objectives: a) To expand the previous published study by analysing the possible enrichment in specific chromatin states (Ernst et al., 2011)., as hypermethylation in enhancers of PD b) To search for genomic correlations between differentially methylated loci and genetic top signal loci in published genome wide association analysis, thus searching genetic risk variants that colocalize with hyper/hypo methylated regions. The latter could be used to prioritize the validation of SNP risk found in these GWAS, and in COURAGE project. Thus, the possibility is now open to model the potential environmental factors found in WP2, in ipsc)-derived dopaminergic neurons. 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 a 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. In 2012, CIBERNED and CIEN Foundation joined this Committee to participate actively in the design of the future COEN scientific strategy. Both institutions are represented by Dr. Miguel Medina, CIBERNED Deputy Scientific Director and member of the CIEN Foundation Scientific Advisory Board. During 2015 the French Agence Nationale de la Recherche (ANR) has also been acknowledged as a new COEN member CIBERNED Annual Report / 248

249 Current COEN members are: Canadian Institutes of Health Research (CIHR) 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 (MESRS), Slovakia CIBERNED-Fundación CIEN, Spain Agence Nationale de la Recherche (ANR), France 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. The first phase of the COEN initiative began at the end of 2010 and was intended to establish common resources and methodological approaches to support future studies. Some of the key issues addressed have been: the refinement and validation of cellular and animal disease models; the development of new measures to define patient subgroups for clinical trials; the identification of new biomarkers to support translational research; the development and harmonization of cognitive test batteries for diagnosis and follow-up of disease progression; and the establishment of common computer platforms to improve 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 a total amount of 5.5 million (of which Spain has provided 600,000 ) in this call to establish an innovative and progressive research program to address the major challenges in this field. The call was 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. The approved projects involving CIBERNED have been completed during 2016: 2016 CIBERNED Annual Report / 249

250 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). The NEURO-MIR Consortium aims to explore the role of micro RNAs (mirnas), molecules capable of controlling gene expression and sensitive to metabolic or environmental stress, in different neurodegenerative diseases (Alzheimer s disease -AD-, frontotemporal dementia -FTD- and amyotrophic lateral sclerosis -ALS-), which share common disease pathways as the hyperexcitable state, defects in protein quality control and axonal transport defects. Given their unique biological function and in vivo stability, mirnas show up as one of the most interesting areas for drug and biomarker development, as well as potential key contributors to the disease progression. Pilot data from partners of this Consortium have already shown neuroprotective effects of silencing a mirna, namely mir-134. In order to achieve the main aim of this project, 4 Workpackages were proposed, from which we have participated in three: Workpackage 1: Identify and interrogate the biological function of AD, FTD and ALS-associated mirnas We have generated animals for the immunoprecipitation of Ago2, a protein required to bring together the mirna and messenger RNA target, in hippocampal samples from control mice, VLW mice that overexpress the microtubule-associated protein tau with 3 mutations found in FTDP-17 (frontotemporal dementia with parkinsonism associated to chromosome 17) patients and Tau-KO mice that don t express tau (Partner 4). Subsequently, Partner 1 evaluated the mirna profile in this samples with OpenArray chips. In addition, Partner 4 generated samples from P301S mice that express tau with a mutation that give rise to a more severe phenotype. This samples will be used for validation studies. The results of this study revealed the presence of 155 mirnas associated to Ago2, an enrichment of mirnas in VLW animals and a decrease in mirnas in Tau-KO animals. Validation of results: mir-134, that controls hyperexcitability, has been validated in two AD models: mice that overexpress tau and mice that overexpress GSK-3/tau (generated by Partner 4). An enrichment of its levels has been detected. In addition, it is not detectable in Tau-KO animals. It is overexpressed in AD brain as well. mir-22, which controls inflammation, is reduced in GSK-3/tau mice an in AD brain. Workpackage 2: Deliver a systems biology platform for the modelling of mirna biological function in neurodegenerative disorders (without Partner 4 participation). Workpackage 3: Develop strategies for the delivery of antagomirs/mirnas as therapeutics. Antagomir (stable locked nucleic acid-modified oligonucleotides, capable of blocking mirnas) delivery has been optimized in animal models of neurodegenerative diseases. Workpackage 4: Test the role of mirna network changes as biomarker for disease pathogenesis and therapeutic efficacy. Plasma samples from patients presenting mild or severe Alzheimer s disease pathology (provided by Partner 4) were analyzed with OpenArray chips. Subsequently, an additional set of samples from control, dementia and severe AD individuals was provided. The results pointed out that: 2016 CIBERNED Annual Report / 250

251 10 mirnas are overexpressed in dementia, 17 in AD and 4 in both conditions. These mirnas include mir-139-5p and mir mirnas are downregulated in dementia, 28 in AD and 13 in both conditions. In addition, tear drops (provided by Partner 4) from AD patients (control, dementia and AD) are being analyzed with OpenArray chips. Additional results (Partner 4): Given their high expertise in the study of Huntington s disease (HD), Partner 4 has performed a pilot study in HD animal models. mir-22 levels are reduced in the striatum (main brain structure affected in this disease) of presymptomatic animals and increased in symptomatic animals. Likewise, Partner 4 has found alterations in MAP2 splicing in HD that could be a consequence of mirnas generated from intronic sequences and that could silence the expression of their host genes in an autoregulatory loop (Cabrera and Lucas, submitted). Regarding dementia animal models, Partner 4 has described how tau alteration (levels and/or 4R/3R ratio) is sufficient for the appearance of Tau Nuclear Rods (TNRs), a new histopathological hallmark previously described by the group in HD, in the mouse model of FTDP-17 called P301S (Fernández-Nogales et al, submitted). Industrial engagements and new collaborations (Partner 4) We have now started a pilot study in Alzheimer disease patients using newly developed biosensors to identify blood purines as biomarkers for different disease stages. Participants: Partner 1, Partner 4 and Sarissa ltd, Coventry, UK ( Biotalentum, Hungary: this company has generated human-induced pluripotent stem cell (hipsc) lines from healthy control individuals as well as from patients suffering from Alzheimer s disease. It is a partner in a recently submitted grant application to the MSCA-ITN-ETN European Training Networks (coordinated by RCSI). During 2015, a second call for Pathfinder projects was resolved in December. The approved project involving CIBERNED, and active during 2016, is described below: Protection of neurons in vitro and in vivo from Synuclein toxicity by molecular tweezers Project partners: Erwan Bezard (Coordinator, France), Richard Wade-Martins United Kingdom), Carlos Matute (CIBERNED). Parkinson disease (PD) is hallmarked by the deposition of aggregated α-synuclein (α-syn) species into intra-neuronal Lewy bodies (LB) and Lewy neurites (LN) inclusions while in multiple system atrophy (MSA) α-syn aggregates form oligodendroglial inclusions. The development of therapeutic strategies to block cell death in PD and MSA has been limited by a lack of understanding of the mechanisms driving neurodegeneration. However, increasing evidence of multiple pivotal roles of α-syn in the pathogenesis of PD and MSA has led to consider the therapeutic potential of several strategies aimed at reducing α-synuclein toxicity. Our objective is to build upon the unique experimental cell (Primary rodent neurons and glial cells - Human neurons derived from PD patients) and animal models of PD (SNCAOVX transgenic mouse and human PD-derived α-syn seed-injected mouse) and MSA (PLP-SYN transgenic mouse and human MSA-derived α-syn seed-injected mouse), developed by this consortium, for testing whether the prototypical molecular tweezer CLR01, shown to inhibit aggregation and toxicity of other amyloidogenic proteins, could protect neurons in vitro and in vivo from PD / MSA-associated synaptotoxicity and reduce PD and MSA s disease like α-syn pathology. The project will thus (in) validate the molecular tweezers CLR01 for PD and/or MSA indications CIBERNED Annual Report / 251

252 International Congress for Research and Innovation in Neurodegenerative Diseases (CIIIEN) During 21st to 23rd September 2016, it was held in Alicante, Spain the Fourth International Congress on Research and Innovation in Neurodegenerative Diseases (CIIIEN), promoted by the Queen Sofia Foundation in collaboration with CIEN Foundation and CIBERNED. The main objective of CIIIEN is providing a forum in which to share progress and information of interest on neurodegenerative diseases among the scientific community. CIIIEN was created in 2013 and consolidates the merger of the two major scientific conferences on neurodegenerative diseases in general and Alzheimer s disease in particular, organized in Spain: the International Symposium Advances in Alzheimer s Disease, promoted annually by the Queen Sofia Foundation and CIEN Foundation, and the 10th CIBERNED Scientific Forum, which brought together every year more than the 58 research groups constituting the CIBER in 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 fourth edition of CIIIEN was once again chaired by Her Majesty Queen Sofia and the scientific program consisted of an opening session, two plenary sessions and five scientific sessions. Among the invited speakers at the conference can be highlighted some international researchers who 2016 CIBERNED Annual Report / 252

253 are leaders in their field of research such as Bengt Winblad (Karolinska Institute, Stockholm, Sweden-), who spoke about treatments and present and future strategies for Alzheimer s disease; Anthony Schapira (University College of London, UK), which presented the latest advances in the treatment of Parkinson s, Eckhard Mandelkow (DZNE, Bonn, Germany-), who addressed the role of Tau protein in the development of Alzheimer s and other neurodegenerative diseases; or Luc Buèe (Lille University, France-) with his lecture on the experimental models of Tau protein and therapeutic strategies. Moreover, in line with the interest to boost the career of CIBERNED young researchers, the 2015 CIBERNED Young Investigator Award was awarded during the Congress to María del Mar Puigdellívol, who presented the study which has been granted such recognition. Ultimately, this event establishes in its fourth 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 between different institutions is becoming increasingly important to obtain optimum results in research. Other international activities Proyecto: Stimulating intrinsic repair for DMD (SIRD)- E-RARE We aim to assess in this period of the project the nature of the distinct immune cell infiltrates in dystrophic muscle at distinct stages of the disease, and whether Wnt7a is able to modulate them. During the first year of the project, we had treated adult DBA-mdx mice (mdx mice in a DBA background, which show anticipated and enhanced muscle fibrosis (Fukada et al, 2010; Pessina et al, 2015)), with non-carrier-free recombinant Wnt7a protein, and, unexpectedly, we found muscle damage after Wnt7a treatment. Trying to solve this problem, we have repeated these experiments with carrier-free recombinant Wnt7a, kindly provided by Dr. Rudnicki s group in Ottawa. We treated adult DBA-mdx mice with carrier free recombinant Wnt7a protein (R&D Systems, as described in von Maltzah et al., PNAS 2012) via intramuscular injection, with the final aim to analyse the effects on muscle inflammation and fibrosis, and in particular the macrophage and fibro/adipogenic progenitor populations by cell sorting, according to the protocols that were set up in the first year. After 21 days, the muscle treated with Wnt7a showed no significant difference in fiber size (Fig. 1), when compared with control animals (injected with vehicle) and no significant difference in fibrosis extension detected by Sirius Red staining (Fig. 2), both indicating no major impact of the treatment neither in the disease progression nor regenerative capacity. To see if there was an impact at the cellular level, we used our previously established FACS protocol to isolate satellite cells, fibro/adipogenic progenitors (FAPs) and different subpopulations of macrophages from Wnt7a and vehicle treated muscles and we analyzed expression of pro- and anti-inflammatory cytokines in the macrophages and pro-fibrotic markers in the satellite cells and FAPs (Fig. 3, 4 and 5). We did not find any significant differences in those markers. Following the suggestion of the Rudnicky lab, we will repeat the same treatment with carrier free recombinant Wnt7a protein form ebioscience, which appears to have a good efficiency in in enhancing muscle growth and beneficial effects in in vivo experiments. Dystrophic mice are presently being aged for this experiment. As an alternative approach we decided to overexpress Wnt7a via electroporation of a plasmid vector, using a BTX electroporating instrument available in our laboratory. We electroporated the phan-wnt7a expression plasmid (Kuroda et al., PNAS 2012) kindly provided by Dr. Rudnicki, in the tibialis anterior (TA) muscle of DBA-mdx mice of 5-8 months old which show significant levels of fibrosis at this stage. For the electroporation we used the standard conditions 2016 CIBERNED Annual Report / 253

254 working in our hands (10 pulses/ 85 volts, external electroporation with approximately 0,5 cm distance among electrodes). The electroporation efficiency was confirmed by plasmid expression, but morphological and fibrosis analyses by H&E and Sirius red respectively, showed no positive effect neither on fiber size or fibrosis reduction 21 days after overexpressing Wnt7a (Fig. 6 and 7). As before, we isolated different subpopulations of macrophages and analyzed expression of pro- and anti-inflammatory cytokines, but in agreement with the histology, we saw no differences (data not shown). Since our partner Dr. C. Trollet in the laboratory of Dr. Gill Butler-Browne in Paris, has recently subcloned the Wnt7a into another plasmid vector with higher expression levels (pgg2-mwnt7a-ha), we decided to electroporate this plasmid in Extensor digitorum longus (EDL), tibialis and gastrocnemius of mdx mice. We included EDL muscle in this experiments, as this muscle gave better results in electroporation experiments (von Maltzahn et al., PNAS 2012). We detected no expression of the plasmid in neither muscle. We sorted immune cells but did not continue the analysis since there was no Wnt7a expression (data not showed). To rule out any technical limitation of the electroporation strategy, we used both plasmids in parallel to electroporate TA and EDL, with and without hyaluronidase injections to increase the electroporation efficiency. Our results show over-expression of Wnt7a from both plasmids to different extents in the TA, but very low levels of expression in the EDL. In summary, we were unable to efficiently modulate Wnt7a expression levels in the EDL after electroporation and the overexpression in TA does not show tissue degeneration nor fibrosis deposition improvement. As another alternative approach to successfully overexpress Wnt7a, we will use the novel adenoviral Wnt7a protein (AAV-Wnt7a) produced by Dr. C. Trollet in the laboratory of Dr. Gill Butler-Browne, in Paris. Identification of Wnt7a-regulated genes in immune cells and FAPs: To decipher the gene expression programs and potential signaling pathways that might be downstream of Wnt7a in immune cells in dystrophic muscle, we aim to analyze the most affected immune cell populations in vivo. For this purpose, we have defined and optimized the FACS conditions for isolating additional immune cell types and update our protocol to isolate the remaining cell types in the muscle interstitium. From the immune cell compartment we have successfully separated T helper (Th), regulatory Tcells (Treg) and innate lymphoid cells (ILC) (Fig. 8). Limbs from 3 monthold mdx (C57BL/6) mice were isolated, sliced and digested for 2 hours in a solution of DMEM containing 0,08% collagenase IV. After consecutive filtrations on 100 um, 70um and 40 um cell strainers, red blood cells were lysed. Cells infiltrating the muscle were then blocked with an antibody blocking the CD16 and CD32 epitopes and stained with the following antibodies (Lin-: CD11b PE CD11c PE CD8 PE, CD3 PE-Cy7, TCRβ AF488, CD4 APC, CD25 APC- Cy7). Lymphocytes were FACS sorted on a BD FACS ARIA II as following: After exclusion debris and dead cells, myeloid cells, dendritic cells and cytotoxic lymphocytes were removed according to their expression of CD8, CD11c or CD11b (Lin-: CD11c-CD11b-CD8). Lymphoid cells were then divided in innate lymphoid cells (defined as CD3+TCRβ-) and classical T-cells based on their expression of TCRβ (CD3+TCRβ+). Among this last population, we isolated three kind of T-cells based on the expression of CD4 and CD25 (IL-2R): T helper (CD4+ CD25-), doublé negative T-Cells (CD4-CD25-) and regulatory T-cells (CD4+CD25+). Following the same separation approach, we have successfully separated the Mast cells and the Eosinophils from immune cell compartment in dystrophic muscle, characterizing the Mast cells as CD11b- CD45+ and ckit+ and the Eosinophils were defined as CD45+CD11b+ and SiglecF+ (Fig. 9). We have optimized as well the separation of FAPs in regenerating muscle. For this separation limbs from 3 month-old WT (C57BL/6) mice 5 days after muscle injury were isolated, sliced and digested for 2 hours in a solution of DMEM containing 0,08% collagenase IV+ dispase 0,05%. After exclusion of debris, dead cells and hematopoietic cells (CD45+), satellite cells were defined as CD45-CD31-α7+ cells, endothelial cells were defined as CD45- CD31- α7 low to high and FAPS were defined as CD45-CD31-α7- Sca1+ (Fig. 10) CIBERNED Annual Report / 254

255 These FACS conditions will be used to characterize the role of Wnt7a on immune cells and fibrosis in dystrophic muscle, once the conditions for the beneficial effects of Wnt7a are finally optimized. Alzheimer s Association The Alzheimer s Association is a non-for-profit organization that focuses on the care and support for patients with Alzheimer s disease, and also funds research through competitive calls for research projects on Alzheimer s disease. During the year 2016, CIBERNED researchers have received funding from the Alzheimer s Association through 2 research projects: A multicenter, randomized, double-blind, placebo-controlled, 4-arm, 26 week parallel-group study to evaluate the safety, tolerability and anti-inflammatory effect of three oromucosal doses of Sativex in patients with mild cognitive impairment of Alzheimer type or early Alzheimer dementia (Sat-CIEN-02): Sat-CIEN-02 is an academic clinical submitted by Dr. Isidre Ferrer to an open and competitive call of the Alzheimer s Association USA that was approved and funded to be run in Spain during the period September-2016 to October CIBERNED has taking over the management of this project as Sponsor. Sixty patients with cognitive impairment or early dementia of Alzheimer type will be recruited in this clinical trial and they will be randomly assigned to one of four treatment arms: placebo and three doses of active drug. The compound that will be assessed is Sativex, a combination of tetrahydrocannabinol and cannabidiol. The trial will be run in 9 hospitales of Madrid (5), San Sebastián (2), Barcelona y Santander. The trial was submitted on the 7th December 2016 and approved by February 2017 by the Ethic Committee and the Spanish Medicines Agency (AEMPS). Many specialists and institutions will collaborate to carry out this trial; their contracts, organization and logistics are already running and CIBERNED launch a new line of clinical research. The primary end-point of the trial is to prove the safety and tolerability of the cannabinoids in these patients; in addition some hints about their potential therapeutic effect are expected and will be useful for the design of future efficacy studies. The selected doses in accordance with previous experimental animal studies are low and without psychoactive effects. The indication of these drugs in the Alzheimer s disease is based on their modulatory action on the synaptic activity and their potent anti-inflammatory and neuroprotector effect. GSK-3B-and adult neurogenesis. Therapeutic potential for Alzheimer disease (2015-NIRG ): The brains of AD patients show dysregulated GSK-3β activity. Using a murine model, we have demonstrated that GSK-3β overexpression (OE) produce direct effects on hippocampal neurons (morphological and functional alterations similar to those observed in AD patients) and a cascade of indirect events linked to brain inflammation (Llorens-Martín et al, Molecular Psychiatry, 2013). The main objective of this project is to evaluate the contribution of GSK-3β, in absence of brain inflammation, to the AD-like cellular and behavioral alterations, and to test potential therapeutics. We will combine the use of a novel transgenic mouse with different viral vectors expressing a Doxycycline-inducible transactivator element. The innovative strategy developed in order to test the individual contribution of GSK-3β to AD-like behavioral and cellular alterations represents a highly relevant in vivo model in which to study of the involvement of this kinase in AD progression in the absence of brain inflammation. Non-pharmacological strategies, such as environmental enrichment (EE) produce beneficial effects in AD models, and we will examine whether GSK-3β OE limits EE effectiveness CIBERNED Annual Report / 255

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257 Scientific productivity and other activities

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259 Analysis of CIBERNED scientific productivity in 2016 To complete this section we provide a table of bibliometric indicators for 2015 and A quick analysis shows a strengthening of the improved scientific production in 2016 (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 (number of publications overall and within the first decile or quartiles 1 and 2; percentage of publications led by CIBERNED groups within quartiles 1 and 2; number of publications in high impact journals; or number of national and international collaborations led by CIBERNED groups). Table 1. Control panel. It includes publications indexed in PubMed and Scopus Indicator Change(%) (A) Total number of publications1 541,00 568,00 4,99 (B) Number of publications in quartiles (Q) 1 y 2 457,00 467,00 2,19 (C) ) Number of publications in Q1 y Q2 as percentage of total (A) 84,47 82,22-2,67 (E) Number of publications in 1st decile 165,00 174,00 5,45 (F) Number of publications (1st decile) as percentage of total (A) 30,50 30,63 0,44 (G) Number of publications in Q1 (includes 1st decile) 361,00 347,00-3,88 (H) Number of publications in Q1 as percentage of total (A) 66,73 61,09-8,45 (I) Number of publications in Q1 as percentage of total Q1 and Q2 (B) 78,99 74,30-5,94 (J) Number of publications in Q2 96,00 120,00 25,00 (K) Number of publications in Q2 as percentage of total (A) 17,74 21,13 19,06 (L) Number of publications (Q1+Q2) led by CIBERNED groups (1st author or corresponding author) 246,00 265,00 7,72 (M) Number of publications in Q1 and Q2 led by CIBERNED groups as percentage of total (B) 53,83 56,75 5,42 (N) Number of publications (Q1 and Q2) not led by CIBERNED groups (1st author or corresponding author) 211,00 202,00-4,27 (O) Number of publications in Q1 and Q2 not led by CIBERNED groups as percentage of total (B) 46,17 43,25-6,32 (P) Number of publications in Q1 and Q2 in which CIBERNED appears in the affiliation (as percentage of total (B)) 82,49 79,23-3,96 (Q) Number of publications in Q1 and Q2 led by CIBERNED groups in which CIBERNED appears in the affiliation (as percentage of total (L)) 89,43 85,28-4,64 (R) Number of publications in Q1 and Q2 not led by CIBERNED groups in which CIBERNED appears in the affiliation (as percentage of total (N)) 74,41 71,29-4,19 (S) Number of publications in Q1 and Q2 with international groups 208,00 255,00 22,60 (T) Number of publications in Q1 and Q2 with international groups (S) as percentage of total number of publications (B) 45,51 54,60 19,97 1 Number of publications indexed in ISI (Web of Knowledge) CIBERNED Annual Report / 259

260 Indicator Change(%) (U) Number of publications in Q1 and Q2 with international groups, and led by CIBERNED groups 77,00 111,00 44,16 (V) Number of publications in Q1 and Q2 with international groups, and led by CIBERNED groups as percentage of all international publications (S) 37,02 43,5 3 17,59 (W) Number of publications in Q1 and Q2 with other CIBERS 24,00 11,00-54,17 (X) Percentage of publications in Q1 and Q2 with other CIBERS led by CIBERNED groups 41,67 9,09-78,18 (Y) Number Percentage of published in Q1 y Q2 0,00 0,00 - (Z) Percentage of Percentage of led by CIBERNED groups 50,00 40,00-20,00 (A2) Addition of impact factors of journals (Q1 y Q2) in which articles have been published 2797, ,90-5,76 (B2) Average of impact factors of journals (Q1 y Q2) in which articles have been published 5,63 5,11-9,27 (C2) Number of publications in journals with impact factor >15 34,00 36,00 5,88 Number of publications in Q1 and Q2 with 2 CIBERNED groups 40,00 62,00 55,00 Number of publications in Q1 and Q2 with 3 CIBERNED groups 16,00 11,00-31,25 Number of publications in Q1 and Q2 with 4 CIBERNED groups 5,00 1,00-80,00 Number of publications in Q1 and Q2 with 5 CIBERNED groups 3,00 0,00 - Number of publications in Q1 and Q2 with 6 CIBERNED groups 1,00 0,00 - Number of publications in Q1 and Q2 with 7 CIBERNED groups 0,00 0,00 - Number of publications in Q1 and Q2 with 8 CIBERNED groups 1,00 0,00 - Cooperative projects Program 1 70,00 60,00-14,29 Cooperative projects Program 2 156,00 144,00-7,69 Projects with other CIBERs/RETICs/CIEN network 34,00 47,00 38,24 Basic Research-type projects 75,00 70,00-6,67 Translational research-type projects 152,00 134,00-11,84 Projects with other national research groups 99,00 108,00 9,09 Projects with international research groups 103,00 47,00-54,37 Projects with industry 39,00 68,00 74, CIBERNED Annual Report / 260

261 The following are a few figures that summarize some of the main bibliometric indicators in relation to the scientific production of CIBERNED, including the total production (indexed and not indexed in PubMed and Scopus), during the year 2016 and its evolution since 2011: Figure 1. Percentage of indexed CIBERNED 2016 publications distributed by quartiles 2,7% 7,6% 10,1% 29,6% D1 Q1 20,3% Q2 Q3 Q4 59,3% No IF D1: publications in the 1st decile. QI: publications in the 1st quartile. Q2: publications in the 2nd quartile. Q3: publications in the 3rd quartile. Q4: publications in the last quartile. Sin FI: publications for which no impact factor is available CIBERNED Annual Report / 261

262 Figure 2. CIBERNED production by quartiles/1st decile Q1+Q2: sum of publications in 1st and 2nd quartiles. D1: publications in the 1st decile D1 Q1+Q2 Figure 3. Percentage of publications of excellence (1st decile) CIBERNED % 30% 25% 29% 29% 31% 30% 30% 20% 23% 15% 10% 5% 0% CIBERNED Annual Report / 262

263 Figure 4. Number of CIBERNED publications within 1st and 2nd quartiles and 1st decile D1 Q1 Q2 Figure 5. Evolution of the impact factor from 2011 to 2016 IF: impact factor , ,33 5,365 5, , ,722 5,44 5, , ,551 5, , Mean IF CIBERNED Annual Report / 263

264 Figure 6. CIBERNED publications in high impact journals IF > 30 IF IF CIBERNED Annual Report / 264

265 Figure 7. Scientific journals with 5 or more CIBERNED publications in Movement Disorders Journal of Alzheimer's Disease Molecular Neurobiology BMC Neurology Brain Pathology Scien c Reports Autophagy Neurobiology of Aging Journal of the Neurological Sciences Fron ers in Neuroscience PLoS ONE Journal of Neurology Neurology Cerebral Cortex GLIA Neurobiology of Disease Neurology Neuropathology and applied neurobiology Neuroscience European Journal of Neurology Fron ers in Aging Neuroscience Journal of Neurology, Neurosurgery and Psychiatry Medicine Methods in Molecular Biology Oncotarget Revista de Neurologia Biochimica et Biophysica Acta - Molecular Basis of Disease Brain Structure and Func on Fron ers in Molecular Neuroscience Fron ers in Neuroanatomy Journal of Neurochemistry Acta Neuropathologica Clinical Neuropharmacology European Journal of Medicinal Chemistry Journal of Neuroin amma on Molecular psychiatry Neuropharmacology Parkinsonism and Related Disorders Schizophrenia Research CIBERNED Annual Report / 265

266 Figure 8. Most frequent SUBJECT categories (WoS-JCR) in CIBERNED 2016 publications Neurosciences Clinical Neurology Biochemistry Molecular Biology Cell Biology Pharmacology Pharmacy Mul disciplinary Sciences Psychiatry Medicine Research Experimental Pathology Geriatrics Gerontology Oncology Medicine General Internal Biotechnology Applied Microbiology Cell Tissue Engineering Chemistry Medicinal Endocrinology Metabolism Gene cs Heredity Cross-network programs Neurological tissue banks Neurological Tissue Bank and Biological Samples of the IDIBELL Institute of Neuropathology, Bellvitge Hospital (Head: Dr. Isidro Ferrer). CIEN Foundation Tissue Bank for Neurological Research (BT-CIEIN) (Head: Dr. Alberto Rábano). Neurological Tissue Bank (BTN) at the University of Barcelona (Head: Dr. Eduardo Tolosa). Other platforms DNA Analysis Service (Head: Dr. Jordi Pérez Tur). Electron Microscopy Service (Head: Dr. José Manuel García Verdugo). Neuroimaging Service (Head: Dr. Jose Luis Cantero Llorente). Dementia Genetics Spanish Consortium (DEGESCO) (Coordinator: Dr. Jordi Clarimón) CIBERNED Annual Report / 266

267 Events and other activities CIBERNED has organized or participated in a number of events during the year, among which are particularly noteworthy the IV International Conference on Research and Innovation in Neurodegenerative Diseases (CIIIEN) held together with the X Annual CINBERNED Scientific Forum (held in Alicante on September 21st-23rd) and the Meeting with the Associations of patients with Alzheimer s disease in the Region of Valencia (held on September 20th in Alicante within the framework of CIIIEN), as well as other scientific events, outreach and international cooperation activities described below. International Conference on Research and Innovation in Neurodegenerative Disease (CIIIEN) In 2016, the city of Alicante was selected to hold for the tenth time the annual CIBERNED Scientific Forum, essential for the proper functioning of the Center, which allows CIBERNED s main researchers and collaborators to meet every year to discuss their research findings, present new data, and disseminate the most relevant advances in neurodegenerative disease research over the past year. Like previous years, coinciding with World Alzheimer s Day that is celebrated every year on September 21, the Third International Congress for Research and Innovation in Neurodegenerative Diseases (CIIIEN) was held in Alicante between the 21stand 23rd of September This Conference joint together the X CIBERNED Scientific Forum and the 12th annual International Symposium of the CIEN Foundation in collaboration with the Queen Sofia Foundation. A more detailed description of this event is available at the International Relations section of this report. Meeting with patient of Alzheimer s disease associations in the region of Valencia Within the framework of the IV International Congress of Research and Innovation of Neurodegenerative Diseases held in Alicante, there was also a gathering with associations of patients with Alzheimer s disease in the Region of Valencia. The event took place on September 20th during the afternoon at the Museum of Contemporary Art of Alicante-MACA and was promoted by the Queen Sofia Foundation, CIBERNED, the CIEN Foundation and the Valencian Federation of Associations People with Alzheimer (FEVAFA, for its acronym in Spanish). This meeting brought together some experts and researchers, such as Dr. Javier Sáez Valero, Principal Investigator of CIBERNED at the Alicante Neuroscience Institute - Miguel Hernández University; Dr. Jordi Alom, Head of Neurology at the Elche Hospital and specialist in Alzheimer s; the FEVAFA President, Mr. Emili Marmaneu; Dr. Esther López, AFA-Alicante Psychologist-Gerontologist; the AFA- Elche neuropsychologist Ms. Carmen Alonso; and Ms. M. Isabel González Ingelmo, director of the CRE-Alzheimer in Salamanca (IMSERSO). Through a debate within the radio show Cuéntame on the topic Talk about your research in social media and meetings with the Neurodegenerative diseases Patients Family Associations, an appropriate space for reflection and discussion was generated, which promoted the dissemination of results of recent psychosocial research, as well as a deeper knowledge of the ethical and economic aspects related to psychosocial care and research in the context of dementia. The event ended with the exciting performance of the AFA- Valencia Choir Voices of Memory. IV International Symposium on advances in Alzheimer s disease social health research The IMSERSO s State Reference Center (CRE, for its acronym in Spanish) for People with Alzheimer s Disease and other Dementias Care hosted on Thursday June 2nd the IV International Symposium Advances in Social-Health Research in Alzheimer s Disease. Organized by the CRE-Alzheimer in collaboration with the Queen Sofia Foundation and the University of Salamanca General 2016 CIBERNED Annual Report / 267

268 Foundation, the Symposium brought together national and international experts with the aim of generating a space for scientific exchange and dissemination of results in social health research on dementias. The Symposium was attended by Ms. María Ángeles Pérez Muñoz, Managing Director of CIBERNED, who moderated the table Perspectives on Alzheimer s disease and social health research. Excellence in neurodegeneration seminar series During 2016 CIBERNED has continued to carry out the series of lectures Excellence in Neurodegeneration Seminar Series, offering presentations by leading international experts in their areas of research on all aspects of frontier research in neurodegenerative diseases (molecular, cellular, genetic, cognitive, clinical, animal models, biomarkers, imaging, etc.) and related areas. Below there is a list of the seminars held during 2016 within this series: Speaker: Bruce Miller (UCSF), University of California San Francisco (UCSF). Title: Tau protein, a crosstalk in neurodegeneration. Speaker: Bengt Winblad, Karolinska Institute, Stockholm. Title: Alzheimer s Disease - Present and future Treatment Strategies. Speaker: Eckhard Mandelkow, DZNE, Bonn-Germany. Title: Role of Tau protein in Alzheimer Disease and neurodegenerative tauopathies: Structure and cellular functions. Speaker: Luc Buée, University of Lille. Title: Tau experimental models and therapeutic strategies. Speaker: David Pardoe, MRC Technologies Dementia Consortium. Title: Collaborative Drug Discovery Opportunities for Neurodegenerative Diseases - The Dementia Consortium. Speaker: Juan Carlos López, Head of Academic Relations and Collaborations, Roche, New York. Title: Challenges and opportunities in transfer of knowledge from academy to industry. Speaker: Anthony Schapira, UCL, London. Title: The Glucocerebrosidase pathway: a paradigm for pathogenesis and treatment in Parkinson disease. CIBERNED Social Forum On the occasion of World Parkinson s Day CIBERNED, the CIEN Foundation and the Parkinson Extremadura Regional Association organized at the Association s headquarters in Mérida a day dedicated to patients, families, doctors, associations, researchers and all those interested in understanding, studying and research in Parkinson s disease. The event was attended by Dr. José Manuel Fuentes Rodríguez, Principal Investigator in CIBERNED and the Managing Director Ms. María Ángeles Pérez Muñoz. Dementia genetics spanish consortium (DEGESCO) DEGESCO (Spanish Dementia Genetics Consortium) is a nation-wide consortium of scientific and technical nature jointly promoted by eleven founding research centers under the institutional umbrella of CIBERNED. DEGESCO was born in 2013 with the general aim of promoting and strengthen the development of genetic studies in order to understand the genetic architecture of neurodegenerative dementias in the Spanish population through the implementation of 2016 CIBERNED Annual Report / 268

269 collaborative projects and actions among its members. DEGESCO philosophy is inclusive, that is, it is open to the incorporation of new research groups, whether public or private as long as they can prove their capacity and interest to conduct research in genetics of dementia. The stable and long-term scientific relationship between the parties has been established in a Memorandum of Understanding signed by all partners during DEGESCO consists of a structure for coordinating sample repositories that share information and processing protocols. Both its scientific, technical quality and ethical compliance with the Biomedical Research Act (Law 14/2007) and Royal Decree on Biobanks (RD 1716/2011) must characterize samples selected by DEGESCO participants. The transfer of samples between consortium members will be in accordance with the requirements of Royal Decree on Biobanks. During 2015, CIBERNED has continued to participate very actively in the activities of DEGESCO to continue a multicenter study on Alzheimer s disease and related dementias in Spain, as well as the Spanish representation in major international consortia. At the end of the year, a scientific symposium was organized to gather the best national experts on genetics of neurodegenerative diseases to address a key area of research by thoroughly reviewing the genetic aspects of the different types of dementia and their implications for the Spanish population. Since its inception in 2013, the consortium has published the following scientific articles: Dols-Icardo O, Iborra O, Valdivia J, Pastor P, Ruiz A, de Munain AL, et al. Assessing the role of TUBA4A gene in frontotemporal degeneration. Neurobiology of aging Feb; 38: 215 e13-4. PubMed PMID: Epub 2015/12/18. Dols-Icardo O, Nebot I, Gorostidi A, Ortega-Cubero S, Hernandez I, Rojas-Garcia R, et al. Analysis of the CHCHD10 gene in patients with frontotemporal dementia and amyotrophic lateral sclerosis from Spain. Brain: a journal of neurology Dec; 138(Pt 12):e400. PubMed PMID: Epub 2015/07/15. Pastor P, Moreno F, Clarimon J, Ruiz A, Combarros O, Calero M, et al. MAPT H1 Haplotype is Associated with Late-Onset Alzheimer s Disease Risk in APOEvarepsilon4 Noncarriers: Results from the Dementia Genetics Spanish Consortium. Journal of Alzheimer s disease: JAD. 2015; 49(2): PubMed PMID: Epub 2015/10/08. 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 Feb; 35(2):444 e1-4. PubMed PMID: Epub 2013/09/18. 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 Nov; 35(11):2657 e13-9. PubMed PMID: Epub 2014/07/22. Young Investigator Award To promote research excellence, CIBERNED awards a prize annually to an emerging scientific involved in clinical or basic research. In the case of the Young Investigator Award of the Year, candidates must be under 35 years of age, members of a CIBERNED research group and lead author of an article published in a indexed scientific journal during The call remit was announced by to all CIBERNED Principal Investigators and through the Center s website. Once the deadline for applications has passed, seven candidates fulfilling the specifications of both calls in a timely manner were received. For the selection of both Awards, the jury consisted of members of CIBERNED Steering Committee and PIs outside the Committee, 2016 CIBERNED Annual Report / 269

270 which evaluated the scientific quality and impact of the publication as well as the candidate s personal contribution. Upon completion of the evaluation process of the applications received, the CIBERNED Scientific Steering agreed to award the 2015 Young Investigator Award to María del Mar Puigdellívol for her work A role for Kalirin-7 in corticostriatal synaptic dysfunction in Huntington s disease, published in Human Molecular Genetics. Other events University of Extremadura (UMEX) summer course Between 18th and 20th July, 2016 the course Advances in neurobiology and neurodegenerative diseases was held in Cáceres, as part of the XVII edition of the International Summer Courses of the University of Extremadura, led by Dr. José Manuel Fuentes Rodríguez, Principal Investigator of CIBERNED and member of the Department of Biochemistry and Molecular Biology and Genetics of the Faculty of Nursing and Occupational Therapy of the University of Extremadura; and by Dr. Miguel Medina, Deputy Scientific Director of CIBERNED. The course was aimed at students and Health Sciences professionals, with the following goals: 1) To learn about the most current advances at the molecular and cellular level of the Nervous System, with special emphasis on what refers to neurodegenerative diseases, given its increasing importance at the social, health and scientific level. 2) To integrate such knowledge to address issues such as understanding, prevention or treatment. 3) To allow an improvement in the acquisition of skills for future professional development, especially if it has to do with scientific research or clinical practice in relation to these or other Nervous System diseases. 4) To obtain the scientific training in the biomedical field that bridges basic and clinical research. 5) To acquire cross curricular knowledge to ensure a multidisciplinary view of the tasks developed by neuroscientists CIBERNED Annual Report / 270

271 3RD Symposium on Biomedical Research IIBM On April 22, 2016, the Symposium entitled Advances and Perspectives in Neuroscience was held at the Campus of the Faculty of Medicine of the Autonomous University of Madrid. Dr. Antonio Cuadrado and Dr. Teresa Iglesias, both Principal Investigators at CIBERNED were part of the organizing committee. The event emphasized the emergence of new findings that contribute to the understanding of the etiology and pathophysiology of diseases of the nervous system, as well as the development and improvement of therapeutic strategies, and the participation of both basic and clinical researchers to promote translation of new scientific findings into clinical practice. The meeting brought together researchers, postgraduate students and sponsors of the biomedical industry and consisted of guest lectures and chaired poster sessions, including time for discussion both inside and outside sessions. XXXV UPV Summer courses - Rare diseases: a global challenge for the Health System On 23rd and 24th June, the course Rare diseases: a global challenge for the health system was held as part of the XXXV Summer Courses of the University of the Basque Country (UPV, for its acronym in Spanish). This course was headed by Dr. Adolfo López de Munaín, principal investigator of CIBERNED. Its aim was to address the problem of rare diseases (RD) globally by giving voice to the protagonists of the subject, which are diseases and their clinical complexities, patients and their associations, the doctors who care for them and the systems organized around healthcare and research on these diseases. The overall goals of the course were: Define the main healthcare problems that affect RDs from the point of view of the professionals. Define the problem of quantification and clinical and epidemiological monitoring of RRDD: registries and information systems. Analyze the main guidelines that should be included in a comprehensive plan on RDs that integrates the care, research, schooling, working, and socio-economic aspects. Collect opinions on the issue from the point of view of those affected. Analyze and compare the results of the strategies in other countries. Check the status of biomedical research on RDs. Collect the opinion and needs of pharmaceutical companies on the development of drugs to treat these conditions. Address the issue of sustainability of the system with the emergence of new treatments for these diseases CIBERNED Annual Report / 271

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273 Financial report

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275 The Network Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED) is a research organization with its own legal personality, according to Article 6.5 of Law 30/1992 of November 26th, on the Legal Regime of Public Administrations and the Common Administrative Procedure, whose mission is the broadly defined monographic research on neurodegenerative diseases. It is composed of research groups, without physical contiguity, belonging to different Administrations, Institutions and Regions, from the public and private sector with research lines and goals focused on the specific common area of neurodegenerative diseases and coordinating for the achievement of objectives which could hardly be envisaged in a more restricted context of implementation. CIBERNED became part of the State Public Administrative Sector on May 14, Since January 1, 2011, it has been subject to the Public Accounting General Plan, approved by Order EHA/1037/2010, as an accounting standard, taking into account the Adaptation of the PGCP for Public Entities whose expenditure budget are estimated. Its public procurement system is governed by Royal Legislative Decree 3/2011, of November 14, which approves the revised text of the Public Sector Contracts Act. CIBERNED is a virtual Research Center consisting of research groups belonging to different Administrations and Institutions in partnership: the member Principal Investigators work in the institutions to which they belong by participating actively and simultaneously in CIBERNED s own cooperative research agenda. It is therefore the result of the collaboration and association of entities as well as the sum of research groups. In 2016 it consisted of 53 groups, an average of 132 hired staff which up to a total of 137 by December 31, The Scientific Head Office is located at the Center for Molecular Biology Severo Ochoa in Madrid. The headquarters and the Manager Office, are located at Valderrebollo 5 in Madrid within the premises of the Alzheimer Center of the Queen Sofia Foundation. Funding For the execution of the activities and general objectives mentioned above the consortium counts on mainly with funding by the Carlos III Institute of Health as organism promoter. On February 9, 2016, a resolution of the Carlos III Institute of Health was published which grants assistance to CIBER in Neurodegenerative Diseases. Said resolution establishes in its Annex that the Carlos III Institute of Health will transfer the nominative subsidy therein, for a total amount of three million, nine hundred and twenty-three thousand two hundred and forty euros (3,923, ) for current expenditure, and one hundred and sixty three thousand three hundred and eighty euros (163, ) for capital expenditure. To this amount of 4,086,620.00, it is necessary to add the co-financing provided by the Associated Institutions, which in 2016 amounts to 2,606, CIBERNED Annual Report / 275

276 Basis of presentation of the accounts The financial statements have been prepared from the accounting records of the Institution, having applied the existing legal provisions on accounting matters, and especially, to present fairly the assets, financial situation and results of the Institution. Accounts are displayed in the standard model and following the accounting principles suggested in the General Public Accounting Plan. More specifically, the Adjustment Plan Public Accounting for government agencies whose spending budget is an estimate is applied, as approved by the 28 July, 2011 Order of the General Comptroller of the State Administration. These financial statements have been prepared by the Managing Director of the Institution and will be submitted to approval by the Governing Council. The accounting principles and criteria applied for the preparation of these financial statements are summarized in Note 4 of this report. All mandatory accounting principles that have a significant impact on these accounts have been applied. No non-binding accounting principles have been applied CIBERNED Annual Report / 276

277 Balance sheet year 2016 Assets Equity and liabilities A) Non-current assets ,87 A) Equity ,53 I. Intangible assets ,67 I. Contributed equity 0 II. Material assetsl ,03 II. Generated equity ,53 III. Real state investemnts 0 1. Results from previous years ,45 IV. Long-term financial investments in group companies, joint ventures and associates 0 2. Results in the year ,92 V.Long-term financial investments ,17 3. Reserves 0 VI. Debtors and other long-term accounts receivables III. Valuation adjustments IV. Other assets increases pending allocation to results 0 0 B) Current assets ,27 B) Non-current liabilities ,20 I. Assets assigned for sale 0 I. Long-term provisions ,01 II. Inventory 0 II. Long-term liabilities ,19 III. Debtors and other accounts receivable ,83 III. Debts with group companies, joint ventures and associates 0 IV. Short-term financial investments in group companies, joint ventures and associates 0 C) Current liabilities ,41 V. Short-term financial investments 0 I. Short-term provisions ,59 VI. Accrual 0 II. Short-term debts ,68 VII. Cash and cash equivalents ,44 III. Short-term debts with group companies, joint ventures and associates 0 IV. Creditors and other accounts payable ,14 V. Accrual 0 TOTAL ASSETS (A+B) ,14 TOTAL EQUITY AND LIABILITIES (A+B+C) , CIBERNED Annual Report / 277

278 2016 economic assets income statement 1. Income tax and social contributions 2. Transfers and subsidies received 3. Net sales and services 4. Changes in inventory of finished and in progress of manufacturing goods and impairment loss 5. Work by the company for its fixed assets 6. Other income from ordinary management 7. Excesses of provisions A) TOTAL REVENUES FROM ORDINARY MANAGEMENT ( ) 8. Staff costs 9. Transfers and grants awarded 10. Procurements 11. Other ordinary management expenses 12. Depreciation of fixed assets B) TOTAL COSTS OF ORDINARY MANAGEMENT ( ) I Results (surplus or deficit) of ordinary management (A+B) 13. Impairment loss and results on disposals of non-finacial assets and selling state assets 14. Other non-ordinary items II results of non-dinancial operations (I ) 15. Financial income 16. Financial expenses 17. Financial expenses charged to assets 18. Change of fair value of financial assets and liabilities 19. Exchange differences 20. Impairment loss, retirements and disposals of financial assets and liabilities 21. Grants to fund financial operations III Result of financial operations ( ) IV Net surplus (or deficit) in the year (II + III) , , , , , , , , , , ,35-114,88-185, , , , , ,92 (+-) Adjustments to the account of the outcome of the prior year Adjusted profit for the prior year 2016 CIBERNED Annual Report / 278

279 Identification of programs Programs and other activities have the following breakdown of financial resources for their implementation, the description of each, as well as data and information is detailed in the scientific section of this report: Personnel Implementation costs Total Administration, management and coordination , , ,40 Program 1 Alzheimer s disease and other degenerative dementias , , ,11 Program 2 Parkinson s disease and other degenerative motor disorders , , ,77 Deputy Scientific Director / Cross-network plataforms , , ,45 Training / Young Investigator Award / Mobility , ,00 Media service, internationalization, and transfer of knowledge , ,35 COOPERATIVE RESEARCH Cooperative projects, 3rd call , ,90 Cooperative projects, 4th call , , ,11 Cooperative projects, 5th call , , ,60 Cooperative projects, 6th call , , ,76 Cooperative projects, 7th call 62,39 0,00 62,39 COMPETITIVE PROJECTS PI11/ , ,47 COEN Projects , , ,68 COEN Projects ,00 0,00 0,00 BBVA Research Project , , ,31 RTC , , ,39 FJCI , , ,63 JPND-COURAGE-PD 6.255, , ,24 PIE14/ , , ,28 ALZ.ASSOC/Mª LLORENS , , ,70 ALZ.ASSOC/I. FERRER 0, , ,13 E-Rare/ SIRD , , , CIBERNED Annual Report / 279

280 Personnel Implementation costs Total OTHER PROJECTS Biogen Evaluating the potential for Nrf2-4839, ,56 GW PHARMA , , ,82 SYLENTIS-CIBERNED , ,16 AMO PHARMA 126,90 0,00 126,90 Associated institutions agreements ,75 Annual forum ,49 Contributions associated institutions , , , ,25 The data in the table above show that CIBERNED has allocated 2,759, for implementing research programs, which represents 59.80% of the ordinary operating expenses (excluding taxes and without taking into account the contribution of Associated Institutions). Considering the inkind contribution made by the Associated Institutions comprising CIBERNED, that percentage goes up to 38.21%. The amount allocated in 2016 to cover the costs of the internal calls for cooperative projects adds to 731,903.76, representing 10.14%-15.86% of the overall budget (depending on whether or not the contribution from the Associated Institutions is considered). The expenditure executed in the 7th call is minimal since it starts its execution period in December2016. The percentage of expenditure bound for the execution of research programs and calls cooperative projects during 2016, excluding the contribution from the Associated Institutions, accounted for 76.67% of the expenditure for the year (48.35% taking into account the contribution from the Associated Institutions). The expenditure on personnel devoted to stable research structures (groups and collaborative projects) represents 60.81% of total expenditure (not taking into account the assessing of the contribution from the Associated Institutions). Resources allocated to the Administration Department remain one more year similar to previous years ratios, representing 7% of total ordinary expenses, excluding the value of the contribution from the Associated Institutions. Research programs Cooperative projects Personnel Administration 2016 CIBERNED Annual Report / 280

281 Human resources The table below presents the distribution of human resources, grouped into the categories of own and associated staff: Group Principal investigator Institution Associated Staff 101 Cuadrado Pastor, Antonio Universidad Autónoma de Madrid Fariñas Gómez, Isabel Universidad de Valencia Fuentes Rodríguez, José Universidad de Extremadura 1 2 Manuel 104 González Castaño, José Universidad Autónoma de Madrid López Barneo, José Fundación Pública Andaluza para la 14 3 Gestión de la Investigación en Salud en Sevilla 106 Ceña Callejo, Valentín Universidad de Castilla-La Mancha Vicario Abejón, Carlos CSIC Vila Bover, Miquel Fundació Instituto de Recerca del Hospital 10 2 Universitari Vall d Hebrón 110 Pérez Castillo, Ana María CSIC Iglesias Vacas, Teresa CSIC García Verdugo, José Manuel Universidad de Valencia del Río Fernández, José Instituto de Bioingeniería de Cataluña 1 1 Antonio 201 Canela Campos,Enric Isidre Universidad de Barcelona Kulisevsky Bojarski, Jaime Fundació Institut de Recerca del Hospital 9 2 de la Sant Creu i Sant Pau 203 Lanciego Pérez, José Luis Centro de Investigación Médica Aplicada 4 2 (CIME) 204 Moratalla Villalba, Rosario CSIC Obeso Inchausti, José Ángel Hospital de Madrid - HM Rodríguez Díaz, Manuel Universidad de La Laguna Tolosa Sarró, Eduardo Hospital Clínico y Provincial de Barcelona Labandeira García, José Luis Universidad de Santiago de Compostela Pérez Tur, Jordi CSIC Alberch Vié, Jordi Universidad de Barcelona Fernández Ruiz, Javier Universidad Complutense de Madrid Guzmán Pastor, Manuel Universidad Complutense de Madrid Lucas Lozano, José Javier CSIC Naranjo Orovio, José Ramón CSIC Ávila de Grado, Jesús CSIC Camins Espuny, Antonio Universidad de Barcelona CIBERNED Annual Report / 281

282 Group Principal investigator Institution Associated Staff 403 de Felipe Oroquieta, Javier CSIC Matute Almau, Carlos Universidad del País Vasco Rodríguez Álvarez, José Universidad Autónoma de Barcelona Sáez Valero, Javier Universidad Miguel Hernández de Elche Soriano García, Eduardo Universidad de Barcelona Torres Alemán, Ignacio CSIC Trullás Oliva, Ramón CSIC Vitorica Ferrández, Universidad de Sevilla 4 1 Francisco Javier 412 Wandosell Jurado, Francisco CSIC Comella Carnicé, Joan Xavier Fundació Institut de Recerca del Hospital 4 3 Universitari Vall d Hebrón 415 Gutíerrez Pérez, Antonia Universidad de Málaga Carro Díaz, Eva María Fundación para la Investigación Biomédica 10 2 del Hospital Universitario 12 de Octubre 503 Ferrer Abizanda, Isidro Instituto de Investigación Biomédica de 10 2 Bellvitge (IDIBELL) 504 Lleó Bisa, Alberto Fundació Institut de Recerca del Hospital 11 3 de la Sant Creu i Sant Pau 506 López de Ceballos Lafarga, CSIC 0 1 María 507 Pastor Muñoz, María Fundación para la Investigación Médica 9 0 Asunción Aplicada (FIMA) 508 Mengod Los Arcos, CSIC 2 2 Guadalupe 509 de Pedro Cuesta, Jesús Instituto de Salud Carlos III Bullido Gómez-Heras, María Universidad Autónoma de Madrid 2 2 Jesús 511 Cantero Lorente, José Luis Universidad Pablo de Olavide Infante Ceberio, Jon Fundación Marqués de Valdecilla Muñoz Cánoves, Pura Universidad Pompeu Fabra Fernández Chacón, Rafael Universidad de Sevilla Navarro Acebes, Xavier Universidad Autónoma de Barcelona López de Munain Arregui, Instituto de Investigación Sanitaria 30 5 Adolfo Biodonostia Oficina técnica 0 10 TOTAL CIBERNED Annual Report / 282

283 CIBERNED s Human Resources department promotes the existence of a solid critical mass of researchers with the dynamics of providing highly qualified professionals. This allows us to lead a basic, clinical and translational scientific research, being a reference of trust for society, for the scientific community and for public and private institutions at national and international level. Contributing in a significant way to the generation of new knowledge, its application to the practice of care and the business environment is the objective pursued by CIBERNED researchers. The acknowledgments and awards received by CIBERNED investigators constitute a motivating factor to persist in our willingness to promote R & D & I and to translate it into the implementation of cutting-edge projects in translational research, knowledge enhancement, research culture promotion and fostering the transfer of results to the healthcare and production sectors. This effort to advance and consolidate the research culture aims to support and further improve scientific and technological knowledge, teaching and training, encouraging the development of scientific projects. CIBERNED strengthens its collaboration with other public research institutions through framework and cooperation agreements applying in all its actions a philosophy of quality and continuous improvement. In its quest for excellence it demands: Leadership Innovation and focus towards the transfer of results Leaning towards the health problems and interests of the population Basic, clinical and multidisciplinary collaboration Technical training Effective management Integral quality Professional recognition Dissemination of research activities Calls for personnel CIBERNED calls for personnel are carried out following criteria of equality, merit, capacity and publicity. The systematic procedure to cover job vacancies has been approved by the Steering Committee and the Governing Council. This procedure is in agreement with section 6.2 of ISO 9001:2008. All job announcements are posted on the website of the consortium Institutions and in compliance with the obligations acquired with the various centers, members of CIBERNED Governing Council and managers of the Institutions are informed of the call contents. An integrated management system which provides Principal Investigators with a quick and easy way of managing calls for personnel has been implemented since This system through CIBERNED s intranet has allowed streamlining the selection process and resolution of positions available, with a significant reduction of the time between the announcement and resolution of new hirings. The application of this system facilitates rapid coverage of job positions needs, foreseen or unforeseen, as well as improves and streamlines the organizational running by introducing efficiency and quality criteria in the internal Human Resources management according to CIBERNED Quality Plan guidelines established under ISO 9001:2008, thus ensuring to the Principal Investigator the worker incorporation on the desired date. Human Resources policy at CIBERNED directs its selective processes towards the recruitment and retention of talent, allowing the incorporation of highly qualified personnel whose levels of technical and behavioral skills are suited to the job profiles sought. During the year 2016 a total of eight calls for personnel have been published, with 37 job positions posted, with a total of 819 job applications received CIBERNED Annual Report / 283

284 Call year Number of calls Available positions Number of applicants Applicants per position ,32 22,14 All jobs posted are defined with a specific profile, required qualifications, job requirements and roles to be developed space as well as the working hours and location of the workplace. Out of the 37 positions filled-in during 2016, ten people had a PhD degree, eighteen were graduates, and nine were advanced technicians. Category Doctor (PhD) Graduate (BSc, MSc) Advanced technician CIBERNED welcomes its new workers After the selection process carried out by CIBERNED, it is important that the newly hired professional identifies himself/herself with his/her work, with the institution and adapts to it as quickly as possible. With the aim of promoting a quick and optimum integration of workers, all those who start their employment relationship are provided with the Welcome Handbook. This guide contains all information that will be useful in providing an overview of our organization as well as basic guidelines to ease their working relationship. Its content includes a brief presentation of CIBERNED, its origin and objectives, job classification, organization of work, leaves and vacations, a summary of Occupational Risk Prevention (ORP) and templates to handle vacations and ORP. They are also provided with a Safety and Health Handbook, which include information regarding their job position. In order to ensure all employees health and welfare in all activities undertaken at the workplace, CIBERNED provides a favorable work environment to health in accordance with the highest standards of reliability, hygiene and safety. The Human Resources department ensures the safety of each worker and compliance with the Health and Safety at the Workplace protocols, seeing as an opportunity to detect apportunities for improvement in those areas most closely linked to work procedures, professional development, competences, training and communication with researchers CIBERNED Annual Report / 284

285 Jobs posted PhD Graduate Advanced technician Distribution of research groups personnel during 2016 CIBERNED personnel by type of contract in 2016 During 2016 the distribution of personnel by type of contract was 31 hired permanently and 125 part-time Permanent 2016 CIBERNED Annual Report / 285

286 CIBERNED personnel by category in 2016 The distribution of personnel according to their degree was 46 Doctors (PhD), 62 graduates, 4 undergraduates, 38 advanced technicians, and 6 administrative assistants PhD Graduate Undergraduate Advanced technicians 62 CIBERNED personnel by gender in 2016 CIBERNED workforce during 2016 consisted of 113 women and 43 men. 43 Female Male 113 CIBERNED personnel by region in 2016 The geographic distribution of personnel of the research teams by regions was as follows: Basque Country 9 Canary Islands Galicia Extremadura Valencia Navarra Madrid 64 Catalonia 47 Cas le-la Mancha 5 Cantabria 3 Andalusia CIBERNED Annual Report / 286

287 Training program With the aim of promoting and developing programs for the scientific training of researchers, CIBERNED has promoted or participated during 2016 in a wide variety of training activities including lectures, presentations, courses and workshops. On April 7, 2016 CIBERNED participated, in collaboration with the CIEN Foundation, the ISCIII European Projects Office and the Technical University of Madrid, in the course of the Course UPM- Expert-Day ISCIII. On the occasion of the World Parkinson s Day on April 11 CIBERNED, the CIEN Foundation and the Regional Association Parkinson Extremadura organized a conference for patients and families, doctors, associations, researchers and all those interested in the understanding, study and research on Parkinson s disease. Between 18th and 20th July 2016, within the International Summer Courses of the University of Extremadura, CIBERNED collaborated and organized the teaching of a Course on Aging and Neurodegeneration led by Dr. José Manuel Fuentes. In addition, CIBERNED participated in a Symposium titled Bench to Bed Research on the Transcription Factor NRF2 held on October 19th at the Faculty of Medicine of the Autonomous University of Madrid. The Symposium was attended by Drs. Miguel Medina, Alberto Rábano, Antonio Cuadrado and Masayuki Yamamoto, among others. The 5th Iberian Congress on Prions, sponsored by CIBERNED, was held in Barcelona from the 28th to the 29th of November, where lectures and poster communications were presented on the latest advances in the study of diseases caused by prions and related neurodegenerative disorders in humans and animals. On the other hand, a number of programs on continuous training for professional and personal development of the employees have been promoted during Among the courses or training activities carried out, the following can be highlighted: Course on Employment skills Five-month stay at the Karolinska Institute in Stockholm Intra-CIBER secondments Radiation Facility Supervisor Course Course on European Patent Law Altmetrics course: Alternative metrics for scientific evaluation Course on Quality Systems Management Prevention of occupational risks The preventive activities carried out during 2016 have contributed to the proper implementation and application of the Plan for Prevention of Occupational Risks established in CIBERNED. During the period of evaluation, various preventive actions have been developed, as well as training and dissemination activities. The Health Surveillance program, through properly validated procedures, aims tat systematically and regularly detecting the symptoms and early signs of job-related injuries, detecting risk situations, and proposing the necessary preventive measures. The medical examinations performed during 2016 were 91, subject to specific protocols depending on the risks to which the employees are exposed. The protocols applied are: 2016 CIBERNED Annual Report / 287

288 Protocol Biological agents-data Display Screen-Akward Postures-Chemical reagents Data Display Screen Biological agents-screens-akward postures-ionizing radiations Chemicals Data Display Screen-Manual handling of loads Number The study sample consists mainly of women who represent 71.43%. The most frequent age range is years, which is 43.96%. For risk assessment reports, the location of the groups that have been revised and updated is specified below: Province Madrid Santa Cruz de Tenerife Barcelona Workplace of groups assessed Cajal Institute -CSIC Faculty of Medicine, University of La Laguna Institute of Bioingeneering of Catalonia In relation to preventive training referred to article 19 of the Law on Prevention of Occupational Risks, on-site courses and online training of occupational hazards in research laboratory settings have been carried out. On the other hand, in order to ensure effective protection in the area of occupational safety and health, a procedure has been established to deal with situations that could constitute workplace harassment. Finally, the accident claims objectives have been met, with no accident occurring during workdays CIBERNED Annual Report / 288

289 Quality During 2016, renewal of the Certification of the Quality System Management under the UNE-EN ISO 9001:2008 standard in the Management department of CIBERNED-CIEN Foundation and the Management of the Tissue Bank has been successfully passed. The Quality policy seeks guaranteeing and optimizing the processes related to: focus on the external and internal customer, leadership, staff participation, process-based approach, continuous improvement and incorporation of the risk-based process of the new ISO 9001:2015 norm. During this year, the procedures required by the new version of the UNE standard have been developed and the working tools have been adapted, providing greater predictive capacity for the evaluation, administration, elimination and/or mitigation of risks. The following measures are noteworthy: the adoption of a process-based approach, the management of internal and external risks and the planning of quality goals. Quality goals are established annually with the aim of achieving continuous improvement in the processes and obtaining higher levels of satisfaction in the customer service, both externally and internally. The Quality System Management is carried out using the following tools: Audit reports, internal and external. Supplier Evaluation. Customer complaints, suggestions and information. Results of customer satisfaction studies. Evaluation of corrective and preventive actions. Procedures quality indicators. Quality objectives. Internal or external amendments with influence on the Quality System CIBERNED Annual Report / 289

290 Collaboration agreements In 2016 the following agreements have been kept in force by the institution: CIBERNED, CLINIC FOUNDATION FOR BIOMEDICAL RESEARCH, CLiNIC HOSPITAL OF BARCELONA, UNIVERSITY OF BARCELONA AND CSIC IDIBELL-ZECLINICS-CIBERNED CIBERNED-BIOCROSS-ISCIII CIBERNED-BIOCROSS-FCIEN CIBERNED-UNIVERSITY OF BARCELONA UNIVERSITY OF BORDEAUX-UNIVERSITY OF OXFORD- CIBERNED (Dr. Carlos Matute) CIBER-CIBERNED Partner institutions STATE ATTORNEY OFFICE MINISTRY OF JUSTICE Description Confidenciality agreement related to patent Method for the subclassification of patients suffering from Parkinson disease Research collaboration agreement on a zebra fish model of amyotrofic latera sclerosis (ALS) Through expression of mutation XXX Co-ownership agreement related to patent A fast and cost-effective method for apolipoprotein E isotyping as an alternative to APOE genotyping (D. Represa) Collaboration agreement for the implementation of public-private cooperation actions within the call RETOS COLABORACIÓN 2016 Assignment of use of research support equipment needed at the University of Barcelona COEN PROJECT. CONSORTIUM AGREEMENT. Protection of neurons in vitro and in vivo from Synuclein toxicity by molecular tweezers Specific collaboration agreement to govern the participation of CIBERNED research groups in the project Search for biomarkers for the early detection of Alzheimer s disease in the Vallecas project cohort. Project Movement Disorders Legal assistance agreement, formalized on 30/09/2016, adjusting to the novelties derived from Law 40/2015 of October 1st 2016 CIBERNED Annual Report / 290

291 Acquisitions of fixed assets The acquisition of assets made during the year are as follows: Purchase order Equipment description Date Total cost Responsible IP code Analytical balance 60g/0,1 mg internal calibration 25/01/ ,35 José M. Fuentes Accumet AB150 PH BIO KIT (Analytic Phmeter) 60000ANY-maze Video tracking Software 25/01/ /02/ , ,00 José M. Fuentes Jesús Ávila Grey wood cabinet S/P 200/90/45 4 es 25 mm r/ amba 05/02/ ,26 José M. Fuentes HP Notebook 15-ac103ns de 15,6 / Intel i5-5200u/ 8gb 10/02/ ,35 Jesús Ávila imac 21.5 /3.3 QC/16GB/1TB/KB/IEA 10/02/ ,30 Jesús Ávila HP jet-pro color laser printer M452DN+ speaker+webcam 17/02/ ,05 Jordi Pérez Equipment Black side 17 MF keyword + mouse 17/02/ ,00 Rafael Fernández LF 27mp37VQ 27 Monitor 17/02/ ,00 Rafael Fernández WINDOWS 7 PRO 64B 17/02/ ,99 Rafael Fernández Stand table 1,20 m 19/02/ ,24 José M. Fuentes Centrifuge, without rotor 02/03/ ,25 José J. Lucas Centrífuge Rotor FA /03/ ,36 José J. Lucas Samsung 850 EVO solid hard disk int 500gb serial ATA 21/03/ ,95 Jesús Ávila HP Pavillion ns intel I5 equipment 21/03/ ,83 Jesús Ávila Monitor Pavillion 22CW in-ips black 21/03/ ,08 Jesús Ávila HP ProBook 430 G3 Laptop + keyword+ mouse 23/03/ ,28 Servicios generales Thermal Shake Lite 230v (block heater with shaking) 23/03/ ,76 Rafael Fernández Monitor HP 21.5 ProDisplay Mod. P222va LED VGA 04/04/ ,58 Eduardo Tolosa CIBERNED Annual Report / 291

292 Purchase order Equipment description Date Total cost Responsible IP code ROG GL752 VW-T4069T Monitor 07/04/ ,03 José J. Lucas SYSTEM, CHEMIDOC XRS+, IMAGE LAB 13/05/ ,00 Isidro Ferrer Hard Disk with Thunderbolt and USB 3,0 G-Drive 31/05/ ,30 Jesús Ávila LCD Monitor LG 24 MP 58 VQ-P 02/06/ ,70 José J. Lucas LCD Monitor LG 24 MP 58 VQ-P 02/06/ ,70 José J. Lucas Cycloergometer corival 02/06/ ,00 José L. Cantero Software lem CORIVAL CPET 02/06/ ,00 José L. Cantero Body fat monitor Tanita SC- 240 MA 06/06/ ,95 José L. Cantero MacBook pro with 13 retina screen Intel Core i5 06/06/ ,53 Rafael Fernández Monitorized system TCA-3 (PO ) 15/06/ ,24 Antonia Gutiérrez Precision scale Pioneer 2100Gx0,01 G INCAL 17/06/ ,56 José A. Obeso PowerPac HC Power Supply, v (electrophoresis power supply) 20/06/ ,11 Rafael Fernández Anesthesia equipment mod CA-EAC20 (N.S ) + Graefe forceps curved serrated 21/06/ ,68 José A. Obeso Graphic tableta Wacom Intuos pro M 07/07/ ,00 Carlos Matute Removable closure grey/ blue cabinet 12/07/ ,49 Servicios generales Removable closure grey/ blue cabinet 12/07/ ,49 Servicios generales Removable closure grey/ blue cabinet 12/07/ ,49 Servicios generales Removable closure grey/ aluminium cabinet 12/07/ ,49 Servicios generales Tower computer 12/07/ ,01 José M. Fuentes Development of application in intranet purchasing processes 13/07/ ,50 Servicios generales Platform for microhandler of electrophysiology 28/07/ ,27 Rafael Fernández CIBERNED Annual Report / 292

293 Purchase order Equipment description Date Total cost Responsible IP code Digital Just for mouse stereotaxic instrument 04/08/ ,60 Jesús Ávila Cordless micro drill, 220 v + carbide burr drills bits 10/08/ ,98 Jesús Ávila Antivibration table 90x75 cm 18/08/ ,80 José M. Fuentes CO2 incubator shaker New BrunsWick S4li 230 v, 50 Hz 30/08/ ,00 Manuel Rodríguez Monitor LCD LG 24MP58VQ-P 07/09/ ,70 José J. Lucas SMART UPS 2200VAS LCD MODELO SMT22001 (SAI) 12/09/ ,45 Mª López de Ceballos Melamine table grey /10/ ,80 Jordi Pérez Melamine table grey /10/2016 0,00 Jordi Pérez Hard disk 17/10/ ,21 José J. Lucas Hard disk 17/10/ ,21 José J. Lucas Custom computer 17/10/ ,96 José J. Lucas Laminar flow hood Alpina K /10/ ,00 Manuel Rodríguez Monitor 23 LCD LG FULL HD 26/10/ ,56 Jesús Ávila Passive Avoidance Mouse Cage, Step Down+sofware activation code+control unit with touch screen 07/11/ ,35 Miquel Vila Mini shaker 3D 09/11/ ,95 Jesús Ávila Transilluminator 302nm 11/11/ ,60 Jordi Pérez Gel DocumentaTiOn System IMAGER2 VWR 11/11/ ,60 Jordi Pérez Magnetic shaker AGIMATIC-E-SELECTA 15/11/ ,74 Jesús Ávila Magnetic shaker AGIMATIC-E-SELECTA 15/11/ ,74 Jesús Ávila T10 BASIC ULTRA-TURRAX 16/11/ ,57 Carlos Matute WOUNDMAKER 96 TOOL 18/11/ ,70 Mª López de Ceballos Dispersing Elements 10 N - 5 G 24/11/ ,54 Carlos Matute Synology Disk Station DS416 NAS-SATA 02/12/ ,59 Ramón Trullas CIBERNED Annual Report / 293

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