Team 3, headed by Capucine Trollet and Vincent Mouly, is working on the molecular and cellular actors involved in human muscle regeneration, in muscle ageing and in muscular dystrophies including oculopharyngeal muscular dystrophy (OPMD) and Duchenne muscular dystrophy (DMD).

More precisely we are working on RNA metabolism, muscle regeneration, muscle stem cells, and fibrosis, with the final aim of developing innovative therapeutic approaches.

We have a strong expertise on human cellular models (this includes the immortalization facility that we have initiated) – and xenotransplantation (this includes several immunodeficient mouse models and grafting procedures).

Themes currently developed in parallel and in synergy

1. Dissection of the molecular mechanisms relevant to OPMD and muscle ageing
2. Dissection of cellular communication during muscle ageing, fibrosis and regeneration
3. Development of therapeutic approaches.

Team members

Vincent Mouly (DR2 CNRS, co-team leader)
Capucine Trollet (CR1 INSERM, co-team leader)
Mona Bensalah (Doc)
Anne Bigot (CR AIM)
Gillian S Butler-Browne (DR1 INSERM, Emeritus)
Jamila Dhiab (IE)
Justine Flaire (IE)
Yann Frey (M2)
Teresa Gidaro (PH)
Jean Lacau St Guily (PUPH)
Kamel Mamchaoui (IR AIM)
Alix Marhic (M2)
Laura Muraine (IE)
Elisa Negroni (CR AIM)
Jessica Ohana (apprentice)
Sophie Perie (PUPH)
Fanny Roth (Doc)

Contacts

vincent.mouly@upmc.fr
capucine.trollet@upmc.fr

Platform for the immortalisation of human cells and collaboration

Platform for the immortalisation of human cells

Our team has demonstrated some important differences between man and mouse, in the regulation of the proliferation of myogenic precursors. We have chosen to work on human precursors, either in vitro or in vivo, in xenograft models that we have developed in successive European networks. Therefore, in collaboration with W. Wright, we have developed an approach for the immortalisation of human muscle precursors that allows them to retain their ability to differentiate: this has enabled the generation of immortal cell lines derived from patient cells.

This activity, developed with the full support of the AFM as well as the Dutch Duchenne Parent Project NL (DPP-NL) and the Jain Foundation, has resulted in the creation of a platform for the immortalisation of human cells.

This platform has generated over 85 different cell lines, from controls or pathological subjects. Over 20 different pathologies have been immortalised, some with several different mutations.

These cell lines are shared in collaboration with many international teams, either to study physiological or pathophysiological mechanisms, or to test innovative therapeutic approaches. Some of these cell lines have been shared with the pharmaceutical industry.

Figure legend: Examples of immortalised human myoblast cell lines in differentiation, including a DMD cell line suitable for exon skipping (DMD del 48-50)

France-Brazil International Associated Laboratory

Another specific aspect of our team is the development of international collaborations. In addition to numerous collaborations established over many years, we have set up an International Associated Laboratory between INSERM and Sorbonne University for France and FIOCRUZ and the Federal University of Rio de Janeiro for Brazil.

The IAL aims to synergise the expertise in muscle and neuromuscular diseases of French teams and the expertise in immunology and inflammation of the Brazilian teams.

The IAL has already been renewed once, has resulted in 14 publications and a common patent, as well as the exchange of three long-term post-docs, 5 jointly supervised PhDs and many short term exchanges, including visiting professorships in both countries. A common training component (masters and / or doctoral program) is being developed and will involve innovative Biotherapies in the muscle environment.

Figure legend: Human myoblasts in the damaged muscle of an immunodeficient mouse, identified by species-specific antibodies

Key publications

• Malerba A*, Klein P*, Bachtarzi H, Jarmin SA, Cordova G, Ferry A, Strings V, Espinoza MP, Mamchaoui K, Blumen SC, St Guily JL, Mouly V, Graham M, Butler-Browne G, Suhy DA, Trollet C**, Dickson G**. PABPN1 gene therapy for oculopharyngeal muscular dystrophy. Nat Commun. 2017 Mar 31;8:14848.
• Richard P*, Trollet C*, Stojkovic T, de Becdelievre A, Perie S, Pouget J, Eymard B; Neurologists of French Neuromuscular Reference Centers CORNEMUS and FILNEMUS. Correlation between PABPN1 genotype and disease severity in oculopharyngeal muscular dystrophy. Neurology. 2016 88(4):359-365.
• Klein P, Oloko M, Roth F, Montel V, Malerba A, Jarmin S, Gidaro T, Popplewell L, Perie S, Lacau St Guily J, de la Grange P, Antoniou MN, Dickson G, Butler-Browne G, Bastide B, Mouly V, Trollet C. Nuclear poly(A)-binding protein aggregates misplace a pre-mRNA outside of SC35 speckle causing its abnormal splicing. Nucleic Acids Res. 2016 Dec 15;44(22):10929-10945.
• Bigot A, Duddy WJ, Ouandaogo ZG, Negroni E, Mariot V, Ghimbovschi S, Harmon B, Wielgosik A, Loiseau C, Devaney J, Dumonceaux J, Butler-Browne G, Mouly V, Duguez S. Age-Associated Methylation Suppresses SPRY1, Leading to a Failure of Re-quiescence and Loss of the Reserve Stem Cell Pool in Elderly Muscle. Cell Rep. 2015 Nov 10;13(6):1172-1182. doi: 10.1016/j.celrep.2015.09.067.
• Chartier A, Klein P, Pierson S, Barbezier N, Gidaro T, Casas F, Carberry S, Dowling P, Maynadier L, Bellec M, Oloko M, Jardel C, Moritz B, Dickson G, Mouly V, Ohlendieck K, Butler-Browne G, Trollet C, Simonelig M. Mitochondrial dysfunction reveals the role of mRNA poly(a) tail regulation in oculopharyngeal muscular dystrophy pathogenesis. PLoS Genet. 2015 Mar 27;11(3):e1005092.
• Périé S, Trollet C, Mouly V, Vanneaux V, Mamchaoui K, Bouazza B, Pierre Marolleau J, Laforêt P, Chapon F, Eymard B, Butler-Browne G, Larghero J, St Guily JL. Autologous myoblast transplantation for oculopharyngeal muscular dystrophy: a Phase I/IIa clinical study. Mol Ther. 2014 Jan;22(1):219-25.
• Vallese D, Negroni E, Duguez S, Ferry A, Trollet C, Aamiri A, Vosshenirch CA, Füchtbauer EM, Di Santo JP, Vitiello L, Butler-Browne G, Mouly V. The Rag2-Il2rb-Dmd- mouse: a novel dystrophic and immunodeficient model to assess innovating therapeutic strategies for muscular dystrophies. Mol Ther. 2013.