title: Autologous transplantation of muscle precursor cells modified with a lentivirus for muscular dystrophy: human cells and primate models.
abstract: Duchenne muscular dystrophy (DMD) is characterized by the absence of dystrophin. We tested the ability of lentiviral vectors to deliver a transgene into myogenic cells before their transplantation. Enhanced green fluorescent protein (eGFP) transgene was efficiently transferred into cells and eGFP-positive fibers were generated following transplantation. An eGFP-micro-dystrophin transgene under the control of a cytomegalovirus promoter was then transferred with the same viral vector but caused some toxicity to the mono-nucleated cells. We then used instead a muscle creatine kinase promoter. Dystrophin expression was observed in the muscle fibers after the transplantation of such genetically modified cells into mdx and severe combined immunodeficient mice. Micro-dystrophin expression was also observed in monkey muscles a month after allogenic or autologous transplantation of genetically modified myoblasts. Therapeutic exon skipping was induced by infecting myoblasts of a DMD patient, deleted for dystrophin exons 49 and 50, with a lentivirus expressing a U7 small nuclear RNA containing antisense sequences against exon 51. The modification led to correct exon skipping and to the expression of a quasi-dystrophin in vitro and in vivo. These results demonstrate the feasibility of lentiviral-based ex vivo gene therapy for DMD.
authors.affiliation: Unité de Recherche en Génétique Humaine, Centre de recherche du CHUL, CHUQ, Faculté de Médecine, Université Laval, Sainte-Foy, Québec, Canada.
authors.forename: Simon P
authors.initials: SP
authors.lastname: Quenneville
authors.forename: Pierre
authors.initials: P
authors.lastname: Chapdelaine
authors.forename: Daniel
authors.initials: D
authors.lastname: Skuk
authors.forename: Matin
authors.initials: M
authors.lastname: Paradis
authors.forename: Marlyne
authors.initials: M
authors.lastname: Goulet
authors.forename: Joël
authors.initials: J
authors.lastname: Rousseau
authors.forename: Xiao
authors.initials: X
authors.lastname: Xiao
authors.forename: Luis
authors.initials: L
authors.lastname: Garcia
authors.forename: Jacques P
authors.initials: JP
authors.lastname: Tremblay
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004195:Disease Models, Animal; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D000882:Haplorhini; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D016513:Mice, SCID; D032342:Muscle Cells / Q000166:cytology / Q000637:transplantation*; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation / Q000379:methods*
chemical_list: D016189:Dystrophin; D049452:Green Fluorescent Proteins