Aii Symposium: microrna in inflammatory diseases
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1 PROGRAM AND ABSTRACTS Aii Symposium: microrna in inflammatory diseases Wednesday, April 9, 2014 at Center for Molecular Medicine (CMM) L8: 00 lecture hall, Karolinska University Hospital Solna
2 APRIL , 9:00-16:45, AT CMM L8:00 LECTURE HALL, KAROLINSKA UNIVERSITY HOSPITAL SOLNA HOSTS: Ning Xu Landén, Andor Pivarcsi and Enikö Sonkoly, Unit of Dermatology and Venereology, Department of Medicine, 8:30-9:00 Registration 9:00-9:20 MicroRNAs In Inflammatory Diseases Andor Pivarcsi, Department of Medicine, 9:20-10:00 MicroRNAs As Regulators Of Skin Inflammation Ning Xu Landén, Department of Medicine, 10:00-10:40 Therapeutic And Biomarker Potential Of MiRNAs In Cardiovascular Disease Lars Maegdefessel, Cardiovascular Genetics and Genomics Group, Department of Medicine, 10:40-11:00 Coffee Break 11:00-11:40 MicroRNAs: Pathogenic Players And Biomarkers In Multiple Sclerosis Maja Jagodic, Department of Clinical Neuroscience, 11:40-12:20 Tools For mirna & LncRNA Research Peter Mouritzen, VP of R&D, Exiqon A/S, Copenhagen, Denmark 12:20-13:20 Lunch 13:20-14:00 Extracellular MicroRNAs Karin Ekström, Department of Biomaterials, Sahlgrenska Academy at University of Gothenburg; BIOMATCELL VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden 14:00-14:40 MiR-146a Alleviates Chronic Skin Inflammation In Atopic Dermatitis Through Suppression Of Innate Immune Responses In Keratinocytes Ana Rebane, Swiss Institute of Allergy and Asthma Research, University of Zürich, Switzerland; Department of Biomedicine and Translational Medicine, University of Tartu, Estonia 14:40-15:00 Coffee Break 15:00-15:40 Regulation Of MiRNA Pathway During Virus-Infections Tanel Punga, IMBIM, Uppsala University, Sweden 15:40-16:45 Development Of MicroRNAs Therapeutics Is Coming Of Age Sakari Kauppinen, Center for Non-coding RNA Research, Department of Haematology, Aalborg University Hospital and Department of Clinical Medicine, Aalborg University, Copenhagen, Denmark 1
3 MicroRNAs in inflammatory diseases Andor Pivarcsi, Department of Medicine, MicroRNAs (mirnas) are short, non-protein coding regulatory RNAs which regulate gene expression at the posttranscriptional level. Exceeding all expectations, more than 2000 mirna genes have been found only in human in the past decade highlighting the importance of RNA-mediated regulatory networks. MiRNAs have been found to play important roles in development, defining and maintaining tissue- and cell-identify, regulation of signal transduction pathways as well as buffering gene expression and orchestrating the activity of gene networks. The importance of the mirna system has been demonstrated in animal models, in which interference with mirna-activity could lead to diseases, such as developmental abnormalities, cancer or autoimmunity. In the past decade, our knowledge about their involvement in infections and inflammatory diseases, such as viral infections, psoriasis, atopic dermatitis, rheumatoid arthritis and asthma, has increased tremendously. MiRNAs regulate the development of immune system, fine-tune the activity of immune cells, and regulate inflammatory gene networks. The discovery of this novel way of gene regulation has greatly contributed to the understanding of human diseases and may lead to the development of novel diagnostic tools and therapies, some of which are already in human trials. MicroRNAs as regulators of skin inflammation Ning Xu Landén, Department of Medicine, Psoriasis is the most prevalent immune-mediated skin disease in adults and there is no cure for it to date. It shares many pathogenetic similarities with other immune-mediated diseases such as Crohn's disease, rheumatoid arthritis and multiple sclerosis. Thus, it may be used as a model disease to understand disease mechanisms and test new concepts in chronic inflammatory diseases. We previously identified a distinct mirna expression profile in psoriasis skin compared with healthy skin. Our current study aims at understanding putative role(s) of these psoriasis related mirnas in skin, using materials from a large biobank of psoriasis patients in Stockholm. We have shown that in psoriasis skin mir-125b and mir-203 regulate keratinocyte proliferation and differentiation; mir-21 suppresses T cells apoptosis; mir-31 and mir-146a affect the dynamic interaction between keratinocytes and immune cells. In this seminar, we will discuss how mirna research advances our knowledge about psoriasis pathogenesis and the potential to use circulating mirna as biomarker to predict treatment response. Therapeutic and biomarker potential of mirnas in cardiovascular disease Lars Maegdefessel, MD, PhD, Cardiovascular Genetics and Genomics Group at CMM, Department of Medicine, 2
4 Despite tremendous efforts in the traditional approach to reduce mortality of cardiovascular diseases, the identification of the underlying causes, as well as sufficient steering and intervention, remains a great challenge in basic research principles of vascular biology and everyday clinical practice. The traditional method of drug and biomarker design, involving enzymes, cell surface receptors, and other proteins, might not impact the recognition and treatment of cardiovascular diseases to a greater extent, due to the high sensitive nature of the targeted system. In this dismaying scenario, the discovery of an entirely new method of regulation by mirnas, as well as other non-coding RNAs (e.g., long non-coding RNAs and natural antisense transcripts), and their validation as markers and modulators of vascular functionality during pathological conditions, provides new hope for innovative therapy and disease discovery. Research in recent years has assigned mirnas a crucial modulatory role in various physiological as well as pathological processes, including cardiovascular diseases, such as myocardial infarction, stroke, and aortic aneurysm. Our research group is currently focused on exploring the therapeutic and biomarker potential of mirnas, enabling us to combat the growing health burden of different cardiovascular diseases. MicroRNAs: pathogenic players and biomarkers in Multiple Sclerosis Maja Jagodic, Department of Clinical Neuroscience, Center for Molecular Medicine,, Stockholm, Sweden Multiple Sclerosis (MS) is chronic inflammatory demyelinating neurodegenerative disease of the central nervous system (CNS) and the leading cause of non-traumatic neurological disability in young adults. Hundreds of RNAs (mirnas) were found to be dysregulated in various cell types and CNS lesions in MS patients. We characterized mirnas that associate with susceptibility to develop experimental autoimmune encephalomyelitis (EAE), a well-known animal model of MS. Using Illumina sequencing, we identified 43 mirnas that differ between EAE susceptible and resistant strains. Two thirds of the differentially expressed mirnas has previously been associated with MS and other autoimmune diseases, e.g. mir-146a, mir-21, mir-181 and mir-223. One of the most abundant mirna in lymphoid tissue after EAE induction was mir- 21, a well-known oncomir, which has recently been implicated in a number of inflammatory conditions. Interestingly, we found mir-21 to be epigenetically regulated in T cells of MS patients. Our preliminary studies using deficient mice demonstrated dramatic effect of mir-21 on T helper cell differentiation and development of EAE. We also detected mir-21 in cerebrospinal fluid from MS patients together with 60 other mirnas that are currently evaluated as potential biomarkers. Together, these studies add to the growing body of evidence suggesting important role of mirnas as treatment targets and biomarkers in chronic neuroinflammatory and neurodegenerative conditions. Tools for mirna & lncrna research Dr Peter Mouritzen, VP of R&D, Exiqon A/S, Copenhagen, Denmark 3
5 Exiqon has developed a genome-wide LNA -based microrna qpcr platform with unparalleled sensitivity and robustness even in biofluids where microrna levels are extremely low. The platform uses a single RT reaction to conduct full mirnome profiling and allows high-throughput profiling of micrornas without the need for pre-amplification. The qpcr platform is applied in biomarker discovery programs in Exiqon Diagnostics where thousands of biofluid samples have been profiled including serum/plasma and urine. This has allowed determining normal reference ranges for circulating micrornas as well as to identify biomarkers of disease and toxicology. Data will be presented from our development of microrna as biomarkers for early detection of colorectal cancer. In addition to microrna, considerable numbers of non-coding RNA species are being discovered these years and it is becoming clear that these non-coding RNAs play an important role in normal cellular processes and disease. However, the function remains unknown for the majority of these new RNA species. Exiqon has developed highly potent and specific antisense oligonucleotides (ASO) based on LNA for functional analysis of long non-coding RNA, mrna and microrna. In contrast to sirna, the LNA ASOs are able to reach the nucleus where long non-coding RNAs are mainly located. Results will be shown demonstrating efficient knock-down of nuclear located MALAT1 in vivo. About Exiqon Exiqon combines leading-edge scientific expertise in gene expression with our proprietary LNA TM technology. Our products, services and scientific staff enable life science researchers to make groundbreaking discoveries. Moreover, we are addressing the unmet need for a new approach to the diagnosis of cancer. Exiqon markets its products directly on and provides customized advice on analysis of mirnas through Technical Sales Specialists. Exiqon is located in the Medicon Valley area of Copenhagen, Denmark and in Boston, USA. Extracellular microrna Karin Ekström, Department of Biomaterials, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden; BIOMATCELL VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden Exosomes have recently received extensive interest due to their ability of transferring RNA (including mirna) between cells in addition to their potential use in therapy and diagnostics. Exosomes are small packages (~100nm), consisting of a lipid bilayer with surface receptors and proteins and RNA inside. Exosomes have the capacity to deliver messages between neighbor cells or more distant cells via transport in the circulation. Exosomes are released by a variety of cells and can also be found in human body fluids including blood plasma, breast milk, tumor tissues, cerebrospinal fluid and urine. During the last years, we have studied exosomes with the emphasis on their role as messengers in cell-cell communication. Of particular interest is the exosomal communications between cells involved in early inflammation and tissue regeneration. We have examined their specific content and function in the communication between different cells, including stem cells and immune cells. In this presentation, I will give an overview about extracellular mirna, with focus on exosomes and transport of mirna between cells, the function of 4
6 extracellular mirna in healthy and diseased status and the potential application of extracellular mirnas as biomarkers. mir-146a alleviates chronic skin inflammation in atopic dermatitis through suppression of innate immune responses in keratinocytes Ana Rebane, 1,2 Toomas Runnel, 1,3 Alar Aab, 1,2 Julia Maslovskaja, 1,2 Beate Rückert, 1 Maya Zimmermann, 1 Mario Plaas, 4 Jaanika Kärner, 1,2 Angela Treis, 1 Maire Pihlap, 2 Nikoletta Nagy, 5 Lajos Kemeny, 5 Külli Kingo, 6 Mei Li, 7 Mark P. Boldin 8 and Cezmi A. Akdis 1 1 Swiss Institute of Allergy and Asthma Research (SIAF), University of Zürich, Davos, Switzerland 2 Department of Biomedicine and Translational Medicine, University of Tartu, Estonia 3 Institute of Molecular and Cellular Biology, University of Tartu, Estonia 4 Transgenic Technology Core Laboratory, University of Tartu, Estonia 5 Department of Dermatology and Allergology, University of Szeged, Hungary 6 Department of Dermatology and Venereology, Dermatology Clinic, University of Tartu, Estonia. 7 Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique/Institut National de la Santé et de la Recherche Médicale/Université de Strasbourg, Illkirch, France. 8 Department of Molecular and Cellular Biology, Beckman Research Institute, City of Hope, Duarte, CA, USA. Chronic skin inflammation in atopic dermatitis (AD) is associated with elevated expression of pro-inflammatory genes and activation of innate immune responses in keratinocytes. The aim of this study was to investigate the role of mirnas in skin inflammation in AD. We show that mir-146a expression is increased in keratinocytes and chronic lesional skin of AD patients. mir-146a inhibited the expression of numerous pro-inflammatory factors, including IFN-gamma-inducible and AD-associated chemokines CCL5 and CCL8 in human primary keratinocytes stimulated either with IFN-gamma, TNF-alpha or IL-1beta. In a mouse model of AD, mir-146a-deficient mice developed stronger inflammation characterized by increased accumulation of infiltrating cells in the dermis, elevated expression of IFN-γ, CCL5 and CCL8 in the skin and IFN-gamma and IL-1β in draining lymph nodes. Both tissue culture and in vivo experiments in mice demonstrated that mir-146a-mediated suppression in allergic skin inflammation partially occurs through direct targeting of upstream signal transducers of NF-kappaB, CARD10 and IRAK. In addition, human CCL5 was determined as a novel, direct target of mir-146a. Together, our data demonstrated that mir-146a is increased in keratinocytes and skin of AD patients and that this elevated expression, in turn, controls chronic inflammatory processes that are triggered by the activation of NF-kappaB in keratinocytes. Regulation of mirna pathway during virus-infections Tanel Punga, Wael Kamel, Göran Akusjärvi 5
7 Uppsala University, IMBIM, BMC Box 582, 75123, Uppsala MicroRNAs (mirnas) provide a unique level of posttranscriptional gene regulation by modulating a wide range of fundamental cellular processes. Viruses are obligate intracellular parasites, which target cellular macromolecular machineries for their own benefit. Therefore it is not surprising that viruses can modulate mirna pathway in the infected cells. Moreover, viruses especially herpesviruses, also encode for their own viral mirnas needed for optimal amplification and survival of virus in the infected cells. Thus, current evidence indicates that viruses use their encoded mirnas to manipulate both cellular and viral gene expression. Furthermore, viral infection can exert a profound impact on the cellular mirna expression profile by affecting the cellular mirna transport and stability. Taken together, various viruses have developed ways to target different steps in mirna pathway to augment their replication potential. Here we will give an overview about current knowledge of viral mirnas and how viruses modulate mirna pathway. As our laboratory has extensive experiences in human adenovirus research, we will further show how adenoviruses elegantly target cellular mirna pathway. More specifically, we will explain the origin and potential function of adenovirusencoded mirnas, known as mivarnas [1]. 1. Punga T, Kamel W, Akusjärvi G (2013) Old and new functions for the adenovirus virusassociated RNAs. Future Virology 8: Development of microrna therapeutics is coming of age Sakari Kauppinen, Center for Non-coding RNA Research, Department of Haematology, Aalborg University Hospital and Department of Clinical Medicine, Aalborg University, A.C. Meyers Vænge 15, DK-2450, Cph SV, Denmark MicroRNAs (mirnas) function as important post-transcriptional regulators of gene expression in nearly every biological process in the cell, including development, differentiation, cell proliferation and metabolism. Furthermore, emerging evidence implies that mirna dysregulation is prevalent and associated with the pathogenesis and progression of a wide variety of human diseases, including cancer, CNS disorders, viral infections, inflammation, cardiovascular diseases and metabolic disorders. Thus, mirnas have emerged as a new class of promising targets for therapeutic intervention. I will provide an overview of the current strategies for inhibition of mirnas and mirna families using chemically modified antimir oligonucleotides and describe recent advances in the discovery and development of mirna-based therapeutics for the treatment of cardiovascular disease and hepatitis C virus (HCV) infection. 6
8 Stockholm Telefon ki.se
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