Sarcomeric signallig by giant muscle proteins controlling muscle growth and turnover

Lead Research Organisation: King's College London
Department Name: Randall Div of Cell and Molecular Biophy

Abstract

We investigate new mechanisms that control the growth of muscles in response to workload. This will help to understand how muscle loss occurs in patients on intensive care units or with certain genetic muscle diseases.

Voluntary movement of our body, and the pumping functions of the heart require the actions of striated muscles, so called because of their extremely regular striped pattern when viewed in a microscope. These stripes are repeating patterns of molecular machines, called sarcomeres. The sarcomere is organized by the giant protein titin, the largest protein of the human body.

Muscle responds rapidly to changes in use, with disuse leading to muscle loss (called atrophy) and exercise leading to muscle growth (called hypertrophy). These events need to be constantly balanced, and require input from sensors for workload.

We study the role of a protein kinase domain in titin, and the proteins interacting with, in muscle growth and atrophy. We found that the titin kinase can respond to mechanical forces, suggesting it plays a role in the responses of muscle to load. We are studying the mechanism of this mechanosignalling and its disruption in muscle diseases using protein studies and animal models.

Technical Summary

The giant protein titin is crucially required for assembling the contractile structure of the sarcomeres of striated muscle. It combines mechanical, architectural and signalling functions. We propose to use an integrated approach to study the role of the M-band portion of titin, its protein kinase domain and the proteins interacting with it in signalling and maintenance functions during muscle growth and atrophy. Mutations in titin are also associated with cardiac and skeletal myopathies, and we will study titin signalling in these hereditary diseases, as well as in acquired forms of myopathies like acute quadriplegic myopathy.

Publications

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Candasamy AJ (2014) Phosphoregulation of the titin-cap protein telethonin in cardiac myocytes. in The Journal of biological chemistry

 
Guideline Title 219th ENMC International Workshop Titinopathies International database of titin mutations and phenotypes, Heemskerk, The Netherlands, 29 April-1 May 2016
Description Contribution to diagnostic guidelines for titinopathies
Geographic Reach Multiple continents/international 
Policy Influence Type Citation in clinical guidelines
Impact Contributions led to establishment of evidence-based criteria how to define pathogenic TTN mutations, which parameters should be included in a titinopathy database, and which clinical parameters should be preferentially monitored in affected patients for best health outcomes and improved diagnosis.
 
Description Scientific advisor, Association Fran?aise contre les myoathies
Geographic Reach Europe 
Policy Influence Type Participation in advisory committee
Impact The committee advises the Association Fran?aise contre les myoathies for the funding of project grants and fellowships in the area of basic muscle research and myopathy research
 
Description Wellcome Trust Physiological Sciences Funding Committee
Geographic Reach Multiple continents/international 
Policy Influence Type Participation in advisory committee
 
Description citation in clinical reviews in the neurological area
Geographic Reach Multiple continents/international 
Policy Influence Type Citation in clinical reviews
Impact improved genetic diagnostics of early-onset myopathies
 
Description BHF Professorship
Amount £1,511,810 (GBP)
Funding ID CH/08/001 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2008 
End 09/2013
 
Description BHF project grant 2012-2015
Amount £228,302 (GBP)
Funding ID PG/11/127/29322 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2012 
End 10/2015
 
Description Leducq Transatlantic Network
Amount £547,867 (GBP)
Funding ID 11 CVD 04 
Organisation Transatlantic Networks of Excellence in Cardiovascular Research Program 
Sector Academic/University
Country France
Start 05/2011 
End 06/2016
 
Description MRC programme grant 2012-2017
Amount £2,101,238 (GBP)
Funding ID MR/J010456/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 05/2012 
End 04/2017
 
Description Wellcome Trust Collaborative Award in Sciences
Amount £1,164,059 (GBP)
Funding ID 201543/Z/16/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2016 
End 09/2020
 
Description Wellcome Trust project grant
Amount £360,909 (GBP)
Funding ID 093253/Z/10/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2011 
End 12/2013
 
Description intermediate training network
Amount £197,860 (GBP)
Organisation Marie Sklodowska-Curie Actions 
Sector Academic/University
Country Global
Start 11/2009 
End 12/2013
 
Title reagents and diagnostic criteria for Human Myopathy with Early Respiratory Failure 
Description antibodies against Nbr1, MURF2 for immunohistochemistry and Western blotting 
Type Of Material Antibody 
Year Produced 2006 
Provided To Others? Yes  
Impact Research Material used for differential diagnosis for Human Myopathy with Early Respiratory Failure 
 
Title reagents and diagnostic criteria for Salih Myopathy and LGMD2J 
Description antibodies against titin and obscurin, obscurin-like 1 used in immunohistochemistry and Western blotting of patient material 
Type Of Material Antibody 
Year Produced 2008 
Provided To Others? Yes  
Impact Research Material used for differential diagnosis for Human Myopathy with Early Respiratory Failure, Salih Myopathy, tibial muscular dystrophy (TMD) and LGMD2J 
 
Description Biophysics of cytoskeletal proteins 
Organisation Technical University of Munich
Department Department of Biophysics
Country Germany 
Sector Academic/University 
PI Contribution Concept of research project, generation of recombinant proteins and cell biological validation
Collaborator Contribution technical and conceptual advances on single-molecule biophysics
Impact One joint paper: Pernigo et al., Proc. Natl. Acad. Sci. USA, (2010). 107(7): p. 2908-2913. Multi-disciplinary collaboration between the physics team in Munich with expertise in optical trap measurements of protein-protein complexes, and our input in muscle protein biochemistry, analysis of sarcomeric protein interactions, and muscle cell biology.
Start Year 2008
 
Description Biophysics of mechanosignalling 
Organisation Ludwig Maximilian University of Munich (LMU Munich)
Country Germany 
Sector Academic/University 
PI Contribution designed research, provided crucial eagents, wrote paper
Collaborator Contribution developed new biophysical method for analysis of mechanoenzymatics
Impact Publications 18765796, 18550806
 
Description Congenital autophagy disorders 
Organisation Guy's and St Thomas' NHS Foundation Trust
Department Paediatric Neurology
Country United Kingdom 
Sector Hospitals 
PI Contribution Analyse the impairment of the autophagy/lysosomal pathway and muscle ubiquitin ligases in hereditary paediatric disorders with myopathy/cardiomyopathy.
Collaborator Contribution Identifies patients and candidate disease genes
Impact myotubular myopathy trust award
Start Year 2011
 
Description Critical illness myopathy 
Organisation Umea University
Department Clinical Neurophysiology Unit
Country Sweden 
Sector Academic/University 
PI Contribution We analyse muscle samples from a unique rat model for critical illness myopathy developed in Umea for biochemical and cellular changes in sarcomeric mechanosignalling complexes
Collaborator Contribution We receive samples from a unique rat model of critical illness myopathy to assess changes in mechanosignalling complexes in the pathogenesis of this condition
Impact One joint paper: Ochala et al., J. Physiol., (2011). 589(8): p. 2007-26. A multi-disciplinary collaboration that combines clinical expertise in critical illness myopathy in Umea with our expertise in muscle mechanosignalling.
Start Year 2006
 
Description Early-onset Myopathies 
Organisation Pierre and Marie Curie University - Paris 6
Department UMR 787 (Institute of Myology)
Country France 
Sector Academic/University 
PI Contribution We analyse sarcomeric protein mutations leading to early-onset (paediatric) myopathies on the biochemical, biophysical and cell-biological level.
Collaborator Contribution Has provided valuable insight into the human biology of titin kinase and M-band titin by identifying crucial regions affected in early-onset myopathies.
Impact Two joint publications: Carmignac et al., Ann. Neurol., (2007). 61(4): p. 340 - 351. Fukuzawa et al., J. Cell Sci., (2008). 121(11): p. 1841-1851. A multi-disciplinary, translational collaboration that elucidates the disease mechanisms of novel early-onset myopathies.
Start Year 2006
 
Description Mechanoenzymatics 
Organisation Ludwig Maximilian University of Munich (LMU Munich)
Country Germany 
Sector Academic/University 
PI Contribution We expanded our research on mechanoenzymatic kinase signalling in collaboration with the Gaub team at the LMU. We identified new target proteins, expressed those and designed new studies.
Collaborator Contribution publications, shared personnel
Impact Two 2008 publications (Puchner et al., Proc. Natl. Acad. Sci. USA, (2008). 105(36): p. 13385-13390. Puchner et al., Biophys. J., (2008). 95(1): p. 426-434.), and one recent paper (Stahl et al., Biophys. J., (2011). 101(8): p. 1978-86). Ongoing multi-disciplinary work with the Munich physics team, and our expertise and conceptual background in muscle biochemistry and cell biology.
Start Year 2006
 
Title MYBPC3 as diagnostic marker 
Description MYBPC3 as a marker of cardiac ischaemia 
IP Reference GB0908071.4 
Protection Patent application published
Year Protection Granted 2008
Licensed No
Impact marker still in evaluation PATENT APPLICATION TERMINATED 2010
 
Title US Patent Application "Myocardial Markers" 
Description MYBPC3 is an early marker of cardiac ischaemia and could offer more sensitive and earlier detection of heart attacks than other established markers. 
IP Reference US2012156702 
Protection Patent application published
Year Protection Granted 2012
Licensed No
Impact Ha sled to an optimisation programme for antibodies useful in clinical screening
 
Description School Visits- Dulwich College, King's College School, James Allen's Girl School 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Schools
Results and Impact Discussed research topic and biomedical implications with 6th form students

Several pupils mentioned that Cardiovascular research was now a focus of the future study interests. several came for subsequent lab work experience. The schools wish to send more pupils next year.
Year(s) Of Engagement Activity 2009
 
Description school visit- Dulwich College 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Schools
Results and Impact Discussed research topic and biomedical implications with 6th form students

several students mentioned that Cardiovascular research was now a focus of the future study interests. several came for subsequent lab work experience
Year(s) Of Engagement Activity 2009,2011,2012