title: An adeno-associated virus vector efficiently and specifically transduces mouse skeletal muscle.
abstract: Expression of a therapeutic gene in the skeletal muscle is a practical strategy to compensate a patients' insufficient circulating factor. Its clinical application requires a muscle-targeting vector capable of inducing a continuous high-level transgene expression. We modified an adeno-associated virus serotype 2 (AAV2) vector expressing luciferase from the mouse muscle creatine kinase gene promoter-enhancer (Ckm). First, AAVS1 insulator was inserted into the vector genome for transcriptional enhancement. This increased transduction of mouse quadriceps muscle by 11-fold at 4 weeks after intramuscular injection. Second, two capsid modifications were combined (21F capsid): incorporation of a segment of AAV1 capsid to produce a hybrid capsid and substitution of a tyrosine with a phenylalanine. Use of 21F capsid increased muscle transduction further by 18-fold, resulting in 200-fold higher efficacy than that of the unmodified vector. Compared with a vector having human elongation factor 1α promoter which showed similar efficacy in the muscle, this vector having Ckm transduced non-muscle organs less efficiently after intravenous administration. The AAV2 vector composed of the modified genome and capsid provides a backbone to develop a clinical vector expressing a therapeutic gene in the muscle.
authors.affiliation: Pathogen Genomics Center, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Murakami
authors.forename: Takamasa
authors.initials: T
authors.lastname: Takeuchi
authors.forename: Mayuyo
authors.initials: M
authors.lastname: Mori-Uchino
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Mori
authors.forename: Takuma
authors.initials: T
authors.lastname: Fujii
authors.forename: Daisuke
authors.initials: D
authors.lastname: Aoki
authors.forename: Keiichi
authors.initials: K
authors.lastname: Nakagawa
authors.forename: Tadahito
authors.initials: T
authors.lastname: Kanda
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D052283:Creatine Kinase, MM Form / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D015967:Gene Expression Regulation, Viral; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000166:cytology* / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D036022:Capsid Proteins; D052283:Creatine Kinase, MM Form