title: Combination Antisense Treatment for Destructive Exon Skipping of Myostatin and Open Reading Frame Rescue of Dystrophin in Neonatal mdx Mice.
abstract: The fatal X-linked Duchenne muscular dystrophy (DMD), characterized by progressive muscle wasting and muscle weakness, is caused by mutations within the DMD gene. The use of antisense oligonucleotides (AOs) modulating pre-mRNA splicing to restore the disrupted dystrophin reading frame, subsequently generating a shortened but functional protein has emerged as a potential strategy in DMD treatment. AO therapy has recently been applied to induce out-of-frame exon skipping of myostatin pre-mRNA, knocking-down expression of myostatin protein, and such an approach is suggested to enhance muscle hypertrophy/hyperplasia and to reduce muscle necrosis. Within this study, we investigated dual exon skipping of dystrophin and myostatin pre-mRNAs using phosphorodiamidate morpholino oligomers conjugated with an arginine-rich peptide (B-PMOs). Intraperitoneal administration of B-PMOs was performed in neonatal mdx males on the day of birth, and at weeks 3 and 6. At week 9, we observed in treated mice (as compared to age-matched, saline-injected controls) normalization of muscle mass, a recovery in dystrophin expression, and a decrease in muscle necrosis, particularly in the diaphragm. Our data provide a proof of concept for antisense therapy combining dystrophin restoration and myostatin inhibition for the treatment of DMD.
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Surrey, UK.
authors.forename: Ngoc B
authors.initials: NB
authors.lastname: Lu-Nguyen
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Surrey, UK.
authors.forename: Susan A
authors.initials: SA
authors.lastname: Jarmin
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge, UK; Current address: AstraZeneca R&D, Discovery Safety, Drug Safety and Metabolism, Alderley Park, Macclesfield, UK.
authors.forename: Amer F
authors.initials: AF
authors.lastname: Saleh
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Surrey, UK.
authors.forename: Linda
authors.initials: L
authors.lastname: Popplewell
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge, UK.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gait
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Surrey, UK. Electronic address: G.Dickson@rhul.ac.uk.
authors.forename: George
authors.initials: G
authors.lastname: Dickson
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D000831:Animals, Newborn; D001120:Arginine / Q000737:chemistry; D003964:Diaphragm / Q000378:metabolism; D004195:Disease Models, Animal; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D015316:Genetic Therapy; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D060172:Morpholinos / Q000235:genetics; D018482:Muscle, Skeletal / Q000378:metabolism; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D009336:Necrosis; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D016366:Open Reading Frames; D010455:Peptides / Q000737:chemistry; D016364:Reading Frames
chemical_list: D016189:Dystrophin; D060172:Morpholinos; D055435:Myostatin; D016376:Oligonucleotides, Antisense; D010455:Peptides; D001120:Arginine