title: Antisense pre-treatment increases gene therapy efficacy in dystrophic muscles.
abstract: In preclinical models for Duchenne muscular dystrophy, dystrophin restoration during adeno-associated virus (AAV)-U7-mediated exon-skipping therapy was shown to decrease drastically after six months in treated muscles. This decline in efficacy is strongly correlated with the loss of the therapeutic AAV genomes, probably due to alterations of the dystrophic myofiber membranes. To improve the membrane integrity of the dystrophic myofibers at the time of AAV-U7 injection, mdx muscles were pre-treated with a single dose of the peptide-phosphorodiamidate morpholino (PPMO) antisense oligonucleotides that induced temporary dystrophin expression at the sarcolemma. The PPMO pre-treatment allowed efficient maintenance of AAV genomes in mdx muscles and enhanced the AAV-U7 therapy effect with a ten-fold increase of the protein level after 6 months. PPMO pre-treatment was also beneficial to AAV-mediated gene therapy with transfer of micro-dystrophin cDNA into muscles. Therefore, avoiding vector genome loss after AAV injection by PPMO pre-treatment would allow efficient long-term restoration of dystrophin and the use of lower and thus safer vector doses for Duchenne patients.
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Peccate
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France.
authors.forename: Amédée
authors.initials: A
authors.lastname: Mollard
authors.affiliation: Université de Versailles St-Quentin, INSERM U1179, LIA BAHN CSM, Montigny-le-Bretonneux, France.
authors.forename: Maëva
authors.initials: M
authors.lastname: Le Hir
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Julien
authors.affiliation: Department of Physiology, Anatomy and Genetics, South Parks Road, Oxford, OX1 3QX, UK.
authors.forename: Graham
authors.initials: G
authors.lastname: McClorey
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, TW20 0EX, UK.
authors.forename: Susan
authors.initials: S
authors.lastname: Jarmin
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, TW20 0EX, UK.
authors.forename: Anita
authors.initials: A
authors.lastname: Le Heron
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, TW20 0EX, UK.
authors.forename: George
authors.initials: G
authors.lastname: Dickson
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France.
authors.forename: Sofia
authors.initials: S
authors.lastname: Benkhelifa-Ziyyat
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France.
authors.forename: France
authors.initials: F
authors.lastname: Piétri-Rouxel
authors.affiliation: Department of Physiology, Anatomy and Genetics, South Parks Road, Oxford, OX1 3QX, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Wood
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France.|NIHR Biomedical Research Centre, Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1EH, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Voit
authors.affiliation: Sorbonne Universités UPMC Univ Paris 06, Inserm, CNRS, Institut de Myologie, Centre de Recherche en Myologie (CRM), GH Pitié Salpêtrière, 105 bd de l'Hôpital, Paris 13, France stephanie.lorain@upmc.fr.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Lorain
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics; D016189:Dystrophin / Q000235:genetics*; D005091:Exons / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage; D006801:Humans; D018101:Mice, Inbred mdx; D060172:Morpholinos / Q000008:administration & dosage*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000473:pathology; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000628:therapy*; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000628:therapy*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage*; D012508:Sarcolemma / Q000187:drug effects / Q000473:pathology
chemical_list: D016189:Dystrophin; D060172:Morpholinos; D016376:Oligonucleotides, Antisense