title: Development and application of a minimal-adenoviral vector system for gene therapy of hemophilia A.
abstract: To achieve efficient delivery and sustained expression of the human factor VIII cDNA in vivo, a minimal-adenoviral (mini-Ad) vector system was developed. The system is composed of a mini-Ad vector with essential cis-elements (less than 1 kb) of the viral genome, an E1-deleted ancillary Ad with packaging attenuation, and an E1-complementing production cell line. Based on this system, MiniAdFVIII was generated to deliver a 27 kb expression cassette consisting of a full-length human factor VIII cDNA flanked by human albumin promoter and genomic sequences. The MiniAdFVIII vector mediated expression of functional human factor VIII in HepG2 and 293 cells. A single-dose intravenous injection of 10(11) viral particles in hemophilic mice of MiniAdFVIII produced a sustained high-level expression of human factor VIII (at 100-800 ng/ml up to 369 days) which corrected the FVIII-deficient phenotype. Safety studies of MiniAdFVIII showed that there were no significant toxic effects in mice and dogs after single intravessel doses of up to 3 x 10(11) and 6 x 10(12) viral particles, respectively. Studies for developing the MiniAdFVIII vector with a site-specific integration mechanism and progress in the development of a human factor VIII-tolerized mouse model for pre-clinical studies of MiniAdFVIII are reported. Further pre-clinical studies and product development of MiniAdFVIII for clinical trials are also discussed.
authors.affiliation: GenStar Therapeutics Corporation, San Diego, CA 92121, USA.
authors.forename: W W
authors.initials: WW
authors.lastname: Zhang
authors.forename: S F
authors.initials: SF
authors.lastname: Josephs
authors.forename: J
authors.initials: J
authors.lastname: Zhou
authors.forename: X
authors.initials: X
authors.lastname: Fang
authors.forename: R
authors.initials: R
authors.lastname: Alemany
authors.forename: C
authors.initials: C
authors.lastname: Balagué
authors.forename: Y
authors.initials: Y
authors.lastname: Dai
authors.forename: D
authors.initials: D
authors.lastname: Ayares
authors.forename: E
authors.initials: E
authors.lastname: Prokopenko
authors.forename: Y C
authors.initials: YC
authors.lastname: Lou
authors.forename: E
authors.initials: E
authors.lastname: Sethi
authors.forename: D
authors.initials: D
authors.lastname: Hubert-Leslie
authors.forename: M
authors.initials: M
authors.lastname: Kennedy
authors.forename: L
authors.initials: L
authors.lastname: Ruiz
authors.forename: S
authors.initials: S
authors.lastname: Rockow-Magnone
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D004195:Disease Models, Animal; D004285:Dogs; D005169:Factor VIII / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006467:Hemophilia A / Q000628:therapy*; D006801:Humans; D051379:Mice
chemical_list: D005169:Factor VIII