RNA interference as a therapeutic agent for neuromuscular disease

Lead Research Organisation: University of Oxford
Department Name: Physiology Anatomy and Genetics

Abstract

In many dominant genetic diseases the mutant gene that gives rise to the disease co-exists with a normal copy of the gene, frequently only differing from the normal copy by a single letter (nucleotide base) in its genetic code. Any method that was reliably able to discriminate the mutant copy of the gene from the normal copy (i.e. that was sensitive enough to discriminate as little as a single base difference), and destroy it, thereby leaving only normal copies of the gene present, would have great potential as a novel treatment strategy. Such a method now exists based on a powerful new technology called RNA interference (RNAi). We have studied dominant mutations in the muscle acetylcholine receptor (AChR) gene (that codes for an important neurotransmitter receptor essential for normal functioning of the nerve-muscle junction) that cause a disease called slow channel myasthenic syndrome. In this work we have demonstrated proof-of-concept for using RNAi to selectively silence expression of the mutant AChR genes. We now wish to develop further this very promising approach by; studying and optimizing the RNAi sequence and chemical structure for mutant gene silencing; testing these in muscle cell lines derived from patients with this disorder; investigating the parameters for effective and safe delivery of RNAi molecules to the nerve-muscle junction; and finally undertaking trials of this therapy in animals.

Technical Summary

RNA interference (RNAi) is a powerful new method for gene silencing. We have demonstrated proof-of-principle for allele-specific gene silencing using RNAi, which offers a novel therapeutic approach for treating dominant genetic neurological disorders. We also have encouraging preliminary data on siRNA delivery to muscle in vivo. We therefore hypothesize that selective down regulation of mutant genes associated with neuromuscular junction (NMJ) disease is very likely to lessen the symptoms as well as slow the progression of the disease. This project will systematically investigate specific silencing of pathogenic mutant acetylcholine receptor (AChR) subunits that underlie the neuromuscular disorder, slow channel congenital myasthenic syndrome (SCCMS). We will identify highly potent RNAi molecules designed specifically to down regulate expression of genes associated with SCCMS; optimize the specific silencing of mutant AChR alleles in patient muscle cell lines; investigate and optimize RNAi delivery to the NMJ in vivo (testing both local and systemic RNAi delivery with chemically modified RNAi molecules and using viral expression and delivery); and conduct preclinical studies in a relevant animal model to evaluate its potential for correcting synaptic dysfunction. The ultimate goal of the research programme is to identify effective and safe candidate molecules and delivery systems for clinical application.

Publications


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Burnet PW (2011) D-amino acid oxidase knockdown in the mouse cerebellum reduces NR2A mRNA. in Molecular and cellular neurosciences
Ivanova GD (2008) PNA-peptide conjugates as intracellular gene control agents. in Nucleic acids symposium series (2004)
Muntoni F (2011) Targeting RNA to treat neuromuscular disease. in Nature reviews. Drug discovery
 
Description Action Duchenne
Geographic Reach Europe 
Policy Influence Type Participation in advisory committee
Impact Funding for muscular dystrophy research and care of DMD patients
 
Description European training network
Geographic Reach Europe 
Policy Influence Type Influenced training of practitioners or researchers
Impact Recognition that better training of scientists and clinicians and muscle disease biology and therapy is necessary
 
Description Parkinson's UK Advisory Committee
Geographic Reach UK 
Policy Influence Type Participation in advisory committee
Impact Research funding and research policy
 
Description TREAT-NMD
Geographic Reach Europe 
Policy Influence Type Participation in advisory committee
Impact Clinical trials policy for european neuromuscular disease research
 
Description AFM PROGRAMME
Amount £3,200,000 (GBP)
Organisation French Muscular Dystrophy Association (AFM) 
Sector Charity/Non Profit
Country France, French Republic
Start 04/2011 
End 03/2015
 
Description BBSRC Project Grant
Amount £120,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 01/2007 
End 01/2009
 
Description MRC PROGRAMME
Amount £1,650,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 01/2011 
End 12/2015
 
Description WELLCOME HICF
Amount £2,500,000 (GBP)
Organisation The Wellcome Trust Ltd 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 01/2011 
End 12/2014
 
Description Gait collaboration 
Organisation Medical Research Council (MRC)
Department MRC Laboratory of Molecular Biology (LMB)
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Public 
PI Contribution In vivo work, miRNA work, oligonucleotide work
Collaborator Contribution Materials
Impact 18776238 21062902 20068555 18842625
Start Year 2007
 
Description MDEX CONSORTIUM 
Organisation University College London (UCL)
Department UCL Institute of Child Health
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Pre-clinical evaluation of oligonucleotides for muscular dystrophy therapy
Collaborator Contribution New avenues for experimental work
Impact 19713152
Start Year 2006
 
Description OXFORD ATAXIA CONSORTIUM 
Organisation University of Oxford
Department Nuffield Department of Clinical Neuroscience
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Evaluation of RNAi therapies for SCA7
Collaborator Contribution Avenues for research work
Impact 19789634
Start Year 2008
 
Description Oxford Psychiatry collaboration 
Organisation Medical Sciences Division
Department Department of Psychiatry
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Multiple 
PI Contribution Generation of RNAi silencing vectors for psychiatric research
Collaborator Contribution New research avenues
Impact 20828614
Start Year 2007
 
Description ATAXIA UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Primary Audience Participants in your research and patient groups
Results and Impact TALK TO RESEARCHERS, PATIENTS, POLICY MAKERS

EDUCATION, FURTHER DISCUSSIONS
Year(s) Of Engagement Activity 2008,2009,2010
 
Description Achtion Duchenne Charity 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Primary Audience Participants in your research and patient groups
Results and Impact Advisory activity

Key strategic decisions made
Year(s) Of Engagement Activity 2009
 
Description Action Duchenne Charity 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? Yes
Primary Audience Public/other audiences
Results and Impact Talk to general public and patients with muscle disease

Better public understanding
Year(s) Of Engagement Activity 2007,2008,2009
 
Description Action Duchenne Charity 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? Yes
Primary Audience Public/other audiences
Results and Impact Article

N/A
Year(s) Of Engagement Activity 2008,2009
 
Description Muscular Dystrophy Campaign 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Primary Audience Participants in your research and patient groups
Results and Impact Article

na
Year(s) Of Engagement Activity 2009,2010
 
Description Muscular dystrophy campaign 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Primary Audience Participants in your research and patient groups
Results and Impact Advisory activity

Key strategic decisions
Year(s) Of Engagement Activity 2007,2008,2009
 
Description PARKINSON'S UK 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact INFLUENCING RESEARCH POLICY AND FUNDING DECISIONS FOR PARKINSON'S DISEASE RESEARCH

CHANGES IN POLICY
NEW FUNDING INITIATIVES
Year(s) Of Engagement Activity 2008,2009,2010,2011,2012
 
Description Parliament lobby 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Primary Audience Policymakers/parliamentarians
Results and Impact Public meeting wiht MPs

Better understanding and awareness of muscle disease and muscle disease therapies
Year(s) Of Engagement Activity 2008,2009