3rd EURASNET WP12 IFM Meeting on Alternative and Aberrant Splicing in Neuromuscular and Neurodegenerative Processes

26-27 March, 2009

ICGEB Trieste, Italy

Meeting Report


EURASNET Workshop on Alternative Splicing

The workshop was organised by Prof. Francisco E. Baralle, Eurasnet WP 12 coordinator with the following aims:

· Get together molecular biologist, neurobiologists and clinical neurologists to discuss the impact of RNA processing derangements in neuromuscular and neurodegenerative diseases

· Assess the future strategies for a target design therapy of these pathologies;

· Highlight the need for increased interdisciplinary research in this area;

· Obtain an up-to-date overview of current research in Europe of hnRNPs such as TDP 43 and FUS that have been shown to be involved in pathologies such as ALS and FTLD and are now been described in other neurodegenerative diseases.

The workshop consisted of a series of seminars from EURASNET members and invited speakers. The Workshop was organized as a two-day meeting in order to provide several communal coffee and buffet breaks that allowed the participants to discuss and comment on the attended presentations.

The seminars (Annex I) were designed to provide an introduction to alternative splicing in a variety of neuromuscular and neurodegenerative diseases, both from a clinical to a basic science research point of view.

Clinical presentations. Prof. John Van Swieten and Prof. Christoper Shaw (Erasmus Medical Centre, Rotterdam and MRC Centre for Neurodegeneration Research, London, respctively) gave an overview of the clinical genotype-phenotype correlations in Frontotemporal dementias. In particular, Prof. Chris Shaw also reported his account of his recent discovery, recently published in Science, regarding the involvement of a new hnRNP protein (FUS/TLS) in familial ALS cases. Following these presentations, Maria Grazia Spillantini, from the Univ. of Cambridge, extended these overviews from the FTLD/ALS pathologies to Tau-mediated neurodegenerative diseases, a topic that was also discussed later on by Jean Marc Gallo from the MRC Centre for Neurodegeneration Research, London) with particular emphasis on the analysis of tau mutants. Finally, Manuela Neumann from the Institute of Neuropathology, Zurich, Switzerland gave a lecture on the clinical genotype-phenotype correlations of FTLD with particular attention to her 2006 discovery of TDP-43 involvement in these diseases. Her talk was complemented by Stuart-Pickering Brown (University of Manchester, UK) who addressed the question of additional still unidentified markers underlying the occurrence of FTD diseases in the general population.

Basic science presentations. The predominantly clinical talks shortly summarized in the previous section were complemented by a series of lectures more focused on the potential biomolecular mechanisms of disease. In addition, several lectures also focused on the most recent attempts to actually target these diseases with a series of therapeutic strategies based on correcting aberrant alternative splicing.

With regards to the involvement of TDP-43 in FTD/ALS both Prof. Baralle and Dr. Buratti (ICGEB, Trieste) presented their latest data on TDP-43 research with regards to the control of alternative splicing by TDP-43 and its potential involvement with microRNA control and neurodegeneration, respectively. Similarly, Dr. Fabian Feiguin (ICGEB Trieste) completed these lectures by ICGEB researchers describing his recently developed Drosophila animal model that lacks the homologue of TDP-43 and displays several neuromuscular junction defects. In this respect, of particular interest were also the lectures from Christian Haas and Bettina Schmidt (Ludwig Maximilians University, Munich, Germany) who reported an investigation of the TDP-43 proteolytic processing in neurodegeneration and the attempt to develop a TDP-43 based zebrafish model of the disease. Finally, these talks dealing predominantly with TDP-43 were completed the lecture of Jernej Ule (MRC Laboratory of Molecular Biology, Cambridge, UK) that reported his preliminary data on his UV-crosslinking and immunoprecipitation (CLIP) technique to identify in vivo TDP-43 RNA targets in human brains.

With regards to neuromuscular degeneration, Dr. Davide Gabellini (DIBIT-HSR, Milan, Italy) gave a lecture with regards how recent data from his lab suggest that aberrant alternative splicing could explain the peculiar involvement of specific muscle groups in Facioscapulohumeral Muscular Dystrophy (FSHD) whilst Dr. Antoine Clery (Institute of Molecular Biology and Biophysics, ETH, Zurich, Switzerland) reported his latest NMR structural studies of human tra2beta1 and SF2/ASF, two factors involved in the alternative splicing of SMN.

In parallel, considerable attention has also been paid to potential RNA splicing based therapeutic approaches to neuromuscular diseases. In particular, Dr. Matthew J.A. Wood (University of Oxford, UK) described a splice correction therapy for Duchenne Muscular Dystrophy based on the use of antisense oligos to cause exon skipping in patient's muscle tissue to recover dystrophen protein functionality and Prof. Daniel Schümperli (Institute for Cell Biology, Bern, Switzerland) described the current prospects for a U7 snRNA-based gene therapy in the cure for Spinal Muscular Atrophy.

Reaction to the Workshop

All participants (Annex II) have expressed considerable interest and excitement in the areas covered by the Meeting and all agree that alternative splicing as a potential cause of neurodegeneration is a very appealing future research area. Contacts are currently taking place between many participants to work together on particular alternatively spliced genes and events.

Conclusions

The IFM Meeting fulfilled all of its objectives in raising awareness of the importance of alternative and aberrant splicing occurrence in neurodegenerative diseases. The mix of presentations by basic molecular biologists and clinicians was extremely valuable to identify potential areas of common interest and to identify the most promising future lines of basic research and therapy.


ANNEX I

PROGRAMME OF THE IFM

26 March 2009

“The Clinical Settings in Neuromuscular and Neurodegenerative Processes”

Chairperson: Dr Diana Baralle (Wessex Clinical Genetics Service, Princess Anne Hospital, Southampton, UK)

15:00 Registration

15:30 Prof. Christopher E. Shaw (MRC Centre for Neurodegeneration Research, London, UK)

“Aberrant RNA processing in motor neuron disorders”

16:00 Dr. John C. van Swieten (Erasmus Medical Center, Rotterdam, The Netherlands)

"Genotype-phenotype correlations in hereditary Frontotemporal Dementias"

16:30 Dr. Matthew J.A. Wood (University of Oxford, UK)

“Splice correction therapy for Duchenne Muscular Dystrophy”

17:00 Prof. Maria G. Spillantini (University of Cambridge, UK)

“Mechanisms of tau dysfunction in neurodegenerative diseases”

17:30 Break

18:00 Prof. Daniel Schümperli (Institute for Cell Biology, Bern, Switzerland)

“Prospects for a U7 snRNA-based gene therapy for Spinal Muscular Atrophy”

18:30 Prof. Vincenzo Silani (University of Milan, Medical School, IRCCS Istituto Auxologico Italiano, Milan, Italy)

“New approach to the patient with neurologic disease: locate the gene(s)”

“The Molecular Biology of Neuromuscular and Neurodegenerative Processes” part I

Chairperson: Prof. Francisco Baralle (ICGEB, Trieste, Italy)

19:00 Dr. Jean M. Gallo (MRC Centre for Neurodegeneration Research, London, UK)

“Tau alternative splicing in neurodegeneration: mechanisms and therapeutic strategies”

19:30 Dr. Manuela Neumann (Institute of Neuropathology, Zurich, Switzerland)

“Molecular Neuropathology of TDP-43-proteinopathies”

20:00 Get together buffet

21:00 Bus to hotel


27 March 2009

“The Molecular Biology of Neuromuscular and Neurodegenerative Processes” part II

Chairperson: Dr Fabian Feiguin (ICGEB, Trieste, Italy)

8:00 Bus to ICGEB

8:45 Prof. Stuart Pickering-Brown (University of Manchester, UK)

"Molecular mechanisms underlying Frontotemporal Lobar degeneration"

9:15 Prof. Christian Haass (Ludwig Maximilians University, Munich, Germany)

“Proteolytic processing in neurodegeneration”

9:45 Dr. Antoine Clery (Institute of Molecular Biology and Biophysics, ETH, Zurich, Switzerland)

"NMR structural study of human tra2beta1 and SF2/ASF, two proteins involved in the alternative splicing of

SMN"

10:00 Prof. Francisco Baralle (ICGEB, Trieste, Italy)

“Dissection of TDP43 functional interactions”

10:30 Dr. Jernej Ule (MRC Laboratory of Molecular Biology, Cambridge, UK)

“UV-crosslinking and immunoprecipitation (CLIP) identifies TDP-43 RNA targets in human brain”

11:00 Break

11:30 Dr. Emanuele Buratti (ICGEB, Trieste, Italy)

“Is the interaction between the microprocessor complex and TDP-43 functional?”

12:00 Dr. Isabelle Behm-Ansmant (Université Henri Poincaré, Vandoeuvre les Nancy, France)

“DM1 and DM2 and the MBNL1 splicing factor”

“Cell and Developmental Biology of Neuromuscular and Neurodegenerative Processes”

Chairperson: Dr Emanuele Buratti (ICGEB, Trieste, Italy)

12:15 Dr. Davide Gabellini (DIBIT-HSR, Milan, Italy)

“Alternative splicing could explain the peculiar involvement of specific muscle groups in Facioscapulohumeral Muscular Dystrophy (FSHD)”

12:45 Dr. Fabian Feiguin (ICGEB, Trieste, Italy)

“Genetic analyses of TDP-43 function in vivo”

13:15 Dr. Bettina Schmid (Ludwig Maximilians University, Munich, Germany)

“Analysis of TDP-43 in Zebrafish”

13:45 Lunch

15:00 End of the meeting and bus to hotel


ANNEX II - Participants

Prof. Frédéric Allain

Institute of Molecular Biology and Biophysics

Zürich

Switzerland

Dr Karen Anthony

MRC Centre for Neurodegenerative Research

King's College London, Institute of Psychiatry

Dept. of Clinical Neuroscience

London

UK

Ms Chiara Appocher

Neurobiology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Selma Eréndira

Avendaño Vázquez Neurobiology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Yuna Ayala

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr Diana Baralle

Wessex Clinical Genetics Service

Princess Anne Hospital

Southampton

UK

Prof. Francisco E. Baralle

Director-General and Head, Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Marco Baralle

Biotechnology Developmnet Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr Isabelle Behm-Ansmant

UMR 7567

Faculté des Sciences et Technique

Université Henri Poincaré

Vandoeuvre-les-Nancy

France

Dr Amit Berson

The Hebrew University of Jerusalem

Dept. of Biological Chemistry

Jerusalem

Israel

Dr Ron Bochner

Tel-Aviv University

Sackler Faculty of Medicine

Dept. of Molecular Genetics and Biochemistry

Tel-Aviv

Israel

Dr Dario Bossi Studio Medico

Trieste

Italy

Dr Christiane Branlant

Laboratoire AREMS, UMR 7567, CNRS-UHP

Faculté des Sciences et Technique

Université Henri Poincaré

Vandoeuvre-les-Nancy

France

Dr. Mauricio Budini

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Emanuele Buratti

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Yari Carlomagno

Human Genetics Laboratory

Department of Medical Sciences

University of Eastern Piedmont "A. Avogadro"

Novara

Italy

Dr. Antoine Clery

Institute of Molecular Biology and Biophysics

Zürich

Switzerland

Dr Claudia Colombrita

Dept. of Neuroscience

IRCCS Istituto Auxologicao Italiano

Università degli Studi di Milano

Cusano Milanino (MI)

Italy

Dr. Lucia Corrado

Human Genetics Laboratory

Department of Medical Sciences

University of Eastern Piedmont "A. Avogadro"

Novara

Italy

Dr. Luisa Costessi

Mouse Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Andrea D'Ambrogio

Proteomics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Laura De Conti

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr Dorothee Dormann

Ludwig Maximilians University

Adolf-Butenandt-Institute

Department of Biochemistry

Munich

Germany

Ms Alessia Falanga

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Fabian Feiguin

Neurobiology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Mr Eugenio F. Fernandez

Human Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Davide Gabellini

Division of Regenerative Medicine, San Raffaele Scientific Institute, DIBIT

Milan

ITALY

Dr. Jean-Marc Gallo

MRC Centre for Neurodegeneration Research

Institute of Psychiatry

Department of Clinical Neuroscience

King's College London

UK

Mr Vinay Kumar Godena

Neurobiology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Elisa Goina

Human Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Prof. Christian Haass

Ludwig Maximilians University

Adolf-Butenandt-Institute

Department of Biochemistry

Munich

GERMANY

Dr Hadas Keren

Tel-Aviv University

Sackler Faculty of Medicine

Dept. of Molecular Genetics and Biochemistry

Tel-Aviv

Israel

Ms Raffaella Klima

Neurobiology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr Malgorzata Konieczna

Adam Mickiewicz University

Dept. of Gene Expression

Institute of Molecular Biology and Biotechnology

Poznan

Poland

Dr Pietro Laneve

Institute of Molecular Biology and Pathology, CNR

La Sapienza University

Rome

Italy

Dr Diego Marega

Clinic of Urology

Cattinara Hospital

Trieste

Italy

Dr Nathalie Marmier-Gourrier

UMR 7214

Faculté des Sciences et Technique

Université Henri Poincaré

Vandoeuvre-les-Nancy

France

Dr Alexandra Martins

INSERM U614

Faculté de Médecine ed Pharmacie

Rouen

France

Dr. Andres F. Muro

Mouse Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Ms Mirjana Nedeljkovic

Mouse Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Manuela Neumann

Institute of Neuropathology

Zurich

Switzerland

Dr. Franco Pagani

Human Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Ms Monica Passoni

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr Izabela Pasternak

Adam Mickiewicz University

Dept. of Gene Expression

Institute of Molecular Biology and Biotechnology

Poznan

Poland

Mr Tibor Pastor

Human Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Prof. Stuart Pickering-Brown

Clinical Neurosciences

University of Manchester

UK

Dr Dorota K. Raczynska

Adam Mickiewicz University

Dept. of Gene Expression

Institute of Molecular Biology and Biotechnology

Poznan

Poland

Dr Michela Raponi

University of Southampton

Human Genetics

Southampton

UK

Dr Antonia Ratti

Dept. of Neuroscience

IRCCS Istituto Auxologicao Italiano

Università degli Studi di Milano

Cusano Milanino (MI)

Italy

Prof. Janice Robertson

Centre for Research in Neurodegenerative Diseeases

University of Toronto

Toronto, Ontario

Canada

Dr. Bettina Schmid

Ludwig Maximilians University

Adolf-Butenandt-Institute

Department of Biochemistry

Munich

Germany

Prof. Daniel Schümperli

Institute for Cell Biology

Bern

Switzerland

Prof. Christopher E. Shaw

MRC Centre for Neurodegeneration Research

Department of Clinical Neuroscience

Institute of Psychiatry

King's College London

UK

Prof. Maria Grazia Spillantini

Department of Clinical Neurosciences

University of Cambridge

Centre for Brain Repair

Cambridge

UK

Dr Karol Szafranski

Fritz Lipmann Institute

Institute for Age Research

Genome Analysis

Jena

Germany

Ms Claudia Tammaro

University of Southampton

Southampton General Hospital

Southampton

UK

Dr James Tollervey

MRC Laboratory of Molecular Biology

Structural Studies Division

Cambridge

UK

Mr Enrico Tongiorgi

B.R.A.I.N. Centre for Neuroscience

Dept. of Life Sciences

University of Trieste

Italy

Ms Ifeoma Ubby

Human Molecular Genetics Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Dr. Jernej Ule

MRC Laboratory of Molecular Biology

Cambridge

UK

Dr. John C. van Swieten

Department of Neurology

Erasmus Medical Centre Rotterdam-Dijkzigt

THE NETHERLANDS

Dr Zhen Wang

MRC Laboratory of Molecular Biology

Structural Studies Division

Cambridge

UK

Dr. Matthew J.A. Wood

Department of Physiology

Anatomy and Genetics

University of Oxford

UK

Dr Shangxi Xiao

Centre for Research in Neurodegenerative Diseeases

University of Toronto

Toronto, Ontario

Canada

Ms Paola Zago

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy

Ms Lorena Zubovic

Molecular Pathology Group

International Centre for Genetic Engineering and Biotechnology – ICGEB

Trieste

Italy