PROJECTS

Microtubule proteome and MyoNuclei Domains (MNDs) formation/maintenance during skeletal muscle development

Myonuclei actively position throughout muscular development. Growing evidences support a direct connection between regulation of nuclear positioning/shape, myonuclear domains establishment, microtubule architecture maintenance in muscle fibers and normal function of muscles. Microtubule network is completely redesigned during muscle formation. We hypothesize that differences between proteome associated with microtubules in immature and mature fibers contribute to (1) microtubule reorganization and (2) myonuclei localization. We developed a mass-spectrometry strategy to isolate and analyze those two proteomes using an original in vitro system that allow the formation of pure “mature” muscle fibers. This strategy conducts to the selection of nearly 500 candidates that we are currently investigating using a siRNA screen-approach using both immortalized muscle C2C12 cells and primary murine muscle cells.

Rescue myonuclear domains establishment in centronuclear myopathies with chemical compounds

The incorrect positioning of nuclei in the center of myofibers is a hallmark of a class of muscular diseases called centronuclear myopathies (CNM), which includes myotubular myopathy. We have developed a screening strategy that has the potential to identify new molecules that can rescue nuclear positioning defects observed in centronuclear myopathies. The screening proposed here is expected to lead to novel strategies for the treatment of centronuclear myopathies by reducing muscle weakness caused by central nuclei in diseased muscles.

Mutations identification and implication in the physiopathology of cardiomyopathies

Cardiac arrhythmias are cardiomyopathies that regroup different pathophysiological syndromes such as atrial fibrillation, tachycardia and ventricular fibrillation. Deciphering molecular mechanisms implicated in these cardiomyopathies will help the identification of therapeutics targets and the development of new treatments. We are leading a systematic approach that consists in the identification of mutations in patients with related cardiomyopathies. Using iPSCs (Induced pluripotent stem cells) and CRISPR/Cas9 technology, we are developing cardiomyocytes with identified mutations and decipher in vitro implication of associated genome alterations on the behavior of those cells.

SELECTED PUBLICATIONS

• SMAD6 overexpression leads to accelerated myogenic differentiation of LMNA mutated cells.
  Janin A, Bauer D, Ratti F, Valla C, Bertrand A, Christin E, Chopin E, Streichenberger N, Bonne G, Gahe V, Cohen T, Méjat A. Sci Rep(2018) 8(1):5618.
• Microtubule motors involved in nuclear movement during skeletal muscle differentiation.
  Gache V, Gomes ER, Cadot B Mol Biol Cell (2017) .
• Age-dependent ventricular arrhythmias risk, structural and molecular remodeling in systemic arterial hypertension.
  Scridon A, Puertas RD, Manati W, Fouilloux-Meugnier E, Loizon E, Oréa V, Chapuis B, Julien C, Barrès C, Tabib A, Chevalier P. Mech Ageing Dev (2017) .
• Atrial fibrillation is associated with hypermethylation in human left atrium, and treatment with decitabine reduces atrial tachyarrhythmias in spontaneously hypertensive rats.
  Doñate Puertas R, Meugnier E, Romestaing C, Rey C, Morel E, Lachuer J, Gadot N, Scridon A, Julien C, Tronc F, Chapuis B, Valla C, Janin A, Pirola L, Méjat A, Rome S, Chevalier P. Transl Res (2017) .
• Moving and positioning the nucleus in skeletal muscle – one step at a time
  Cadot B, Gache V, Gomes ER. Nucleus (2015) .
• Nuclear movement during myotube formation is microtubule and dynein dependent and is regulated by Cdc42, Par6 and Par3.
  Gache,V.*, Cadot, B*., Vasyutina, E., Falcone, S., Birchmeier, C., Gomes E. EMBO Reports (2012) .
• MAP and Kinesin dependent nuclear positioning is required for skeletal muscle function.
  Gache,V.*, Metzger,T.*, Xu,M., Cadot, B., Folker,E.S., Richardson, B.E., Gomes E., Baylies. M. Nature (2012) .

FUNDING

• AFM-MyoNeurALP program (2018-2022): Myonuclear domains establishment and maintenance during muscle development.
• ATIP Avenir program (2015-2020): Interplay between cytoskeleton network regulation during muscle development and muscle function.
• Centre de references maladies rares (Cardiologie, Pr. Philipe Chevalier)
• ANR: Identification of injectable and haemostatic hydrogel for guiding and reconstruction of deep wounds
• ALMS: Mutations identification and implication in the physiopathology of cardiomyopathies