1. Dystrophin myonuclear domain restoration governs treatment efficacy in dystrophic muscle
- Author
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Adrien Morin, Amalia Stantzou, Olga N. Petrova, John Hildyard, Thomas Tensorer, Meriem Matouk, Mina V. Petkova, Isabelle Richard, Tudor Manoliu, Aurélie Goyenvalle, Sestina Falcone, Markus Schuelke, Corinne Laplace-Builhé, Richard J. Piercy, Luis Garcia, Helge Amthor, Handicap neuromusculaire : Physiopathologie, Biothérapie et Pharmacologies appliquées (END-ICAP), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Institut National de la Santé et de la Recherche Médicale (INSERM), Royal Veterinary College [London], University of London [London], Université de Versailles Saint-Quentin-en-Yvelines - UFR Sciences de la santé Simone Veil (UVSQ Santé), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Berlin Institute of Health (BIH), Généthon, Immunologie moléculaire et biothérapies innovantes (IMBI), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon, Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Centre Scientifique de Monaco (CSM), Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Institut de Myologie, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Hôpital Raymond Poincaré [Garches], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris-Saclay, Association Française contre les Myopathies, AFM, Agence Nationale de la Recherche, ANR, Deutsch-Französische Hochschule, DFH: CDFA-06-11, We thank Thomas Bestetti, Chloé Dambrune, Karima Relizani, and Cécile Gastaldi for technical assistance. We thank Dr. Feng Zhang for providing the plasmid pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA, and U6::BsaIsgRNA. We also thank Frederic De Leeuw for assistance with multiphoton imaging and Matthieu Dos Santos for preliminary RNA scope experiments. This work was supported by the Association Monegasque contre les Myopathies (AMM, Monaco), the Université Franco-Allemande (CDFA-06-11, UFA), the Association Française contre les Myopathies (AFM, France), and the Agence National de la Recherche (ANR, France).
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Gene Editing ,Multidisciplinary ,[SDV]Life Sciences [q-bio] ,Muscles ,dystrophin-EGFP ,nuclear domain ,Dystrophin ,Muscular Dystrophy, Duchenne ,Mice ,Disease Models, Animal ,Treatment Outcome ,Mice, Inbred mdx ,Animals ,Female ,mdx mouse ,myotendinous junction ,Muscle, Skeletal - Abstract
Dystrophin is essential for muscle health: its sarcolemmal absence causes the fatal, X-linked condition, Duchenne muscular dystrophy (DMD). However, its normal, spatial organization remains poorly understood, which hinders the interpretation of efficacy of its therapeutic restoration. Using female reporter mice heterozygous for fluorescently tagged dystrophin ( Dmd EGFP ), we here reveal that dystrophin distribution is unexpectedly compartmentalized, being restricted to myonuclear-defined sarcolemmal territories extending ~80 µm, which we called “basal sarcolemmal dystrophin units (BSDUs).” These territories were further specialized at myotendinous junctions, where both Dmd transcripts and dystrophin protein were enriched. Genome-level correction in X-linked muscular dystrophy mice via CRISPR/Cas9 gene editing restored a mosaic of separated dystrophin domains, whereas transcript-level Dmd correction, following treatment with tricyclo-DNA antisense oligonucleotides, restored dystrophin initially at junctions before extending along the entire fiber—with levels ~2% sufficient to moderate the dystrophic process. We conclude that widespread restoration of fiber dystrophin is likely critical for therapeutic success in DMD, perhaps most importantly, at muscle–tendon junctions.
- Published
- 2023
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