title: In vivo muscle gene transfer of full-length dystrophin with an adenoviral vector that lacks all viral genes.
abstract: Duchenne muscular dystrophy (DMD) is an important target for gene transfer because of the disease's high frequency and devastating course. To date, adenoviral vector-mediated gene transfer for DMD has been unavailable because (1) adenoviral vectors were unable to accommodate the full-length dystrophin cDNA (14 kb); and (2) adenoviral vectors induced inflammatory reactions in the gene transfer recipient. We addressed both problems with a novel adenoviral vector that contains no viral genes and encodes 28.2 kb of foreign DNA including both the full-length dystrophin cDNA with the muscle creatine kinase promoter for transcriptional control and a lacZ marker gene. This report presents the in vivo expression of dystrophin and beta-galactosidase from this vector in skeletal muscle of the mdx mouse, a mutant mouse that lacks dystrophin. Somatic delivery of the vector by intramuscular injection in 6-day-old mice resulted in the expression of full-length, recombinant dystrophin at the muscle membrane. Dystrophin-associated proteins were restored in muscle fibers expressing recombinant dystrophin. Mdx muscle injected with our vector showed a decrease in the proportion of fibers with nuclei located centrally; centrally placed nuclei in muscle fibers are characteristic of cycles of degeneration and regeneration suffered by dystrophin-deficient muscle tissue. These results are strong evidence that adenoviral vector-mediated full-length dystrophin delivery provides substantial somatic function.
authors.affiliation: Department of Neurology, University of Pittsburgh, PA 15213, USA.
authors.forename: P R
authors.initials: PR
authors.lastname: Clemens
authors.forename: S
authors.initials: S
authors.lastname: Kochanek
authors.forename: Y
authors.initials: Y
authors.lastname: Sunada
authors.forename: S
authors.initials: S
authors.lastname: Chan
authors.forename: H H
authors.initials: HH
authors.lastname: Chen
authors.forename: K P
authors.initials: KP
authors.lastname: Campbell
authors.forename: C T
authors.initials: CT
authors.lastname: Caskey
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016189:Dystrophin / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D005822:Genetic Vectors; D051379:Mice; D018101:Mice, Inbred mdx; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D016189:Dystrophin; D001616:beta-Galactosidase