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Dystrophin myonuclear domain restoration governs treatment efficacy in dystrophic muscle

Authors :
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
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).
Source :
Proceedings of the National Academy of Sciences of the United States of America, Proceedings of the National Academy of Sciences of the United States of America, 2023, 120 (2), ⟨10.1073/pnas.2206324120⟩
Publication Year :
2023
Publisher :
Proceedings of the National Academy of Sciences, 2023.

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.

Details

ISSN :
10916490 and 00278424
Volume :
120
Database :
OpenAIRE
Journal :
Proceedings of the National Academy of Sciences
Accession number :
edsair.doi.dedup.....6109efc7d1cf1b4e7be201f55655c9ba