title: Long-term correction of canine hemophilia B by gene transfer of blood coagulation factor IX mediated by adeno-associated viral vector.
abstract: Hemophilia B is a severe X-linked bleeding diathesis caused by the absence of functional blood coagulation factor IX, and is an excellent candidate for treatment of a genetic disease by gene therapy. Using an adeno-associated viral vector, we demonstrate sustained expression (>17 months) of factor IX in a large-animal model at levels that would have a therapeutic effect in humans (up to 70 ng/ml, adequate to achieve phenotypic correction, in an animal injected with 8.5x10(12) vector particles/kg). The five hemophilia B dogs treated showed stable, vector dose-dependent partial correction of the whole blood clotting time and, at higher doses, of the activated partial thromboplastin time. In contrast to other viral gene delivery systems, this minimally invasive procedure, consisting of a series of percutaneous intramuscular injections at a single timepoint, was not associated with local or systemic toxicity. Efficient gene transfer to muscle was shown by immunofluorescence staining and DNA analysis of biopsied tissue. Immune responses against factor IX were either absent or transient. These data provide strong support for the feasibility of the approach for therapy of human subjects.
authors.affiliation: Department of Pediatrics, University of Pennsylvania Medical Center and The Children's Hospital of Philadelphia, 19104, USA.
authors.forename: R W
authors.initials: RW
authors.lastname: Herzog
authors.forename: E Y
authors.initials: EY
authors.lastname: Yang
authors.forename: L B
authors.initials: LB
authors.lastname: Couto
authors.forename: J N
authors.initials: JN
authors.lastname: Hagstrom
authors.forename: D
authors.initials: D
authors.lastname: Elwell
authors.forename: P A
authors.initials: PA
authors.lastname: Fields
authors.forename: M
authors.initials: M
authors.lastname: Burton
authors.forename: D A
authors.initials: DA
authors.lastname: Bellinger
authors.forename: M S
authors.initials: MS
authors.lastname: Read
authors.forename: K M
authors.initials: KM
authors.lastname: Brinkhous
authors.forename: G M
authors.initials: GM
authors.lastname: Podsakoff
authors.forename: T C
authors.initials: TC
authors.lastname: Nichols
authors.forename: G J
authors.initials: GJ
authors.lastname: Kurtzman
authors.forename: K A
authors.initials: KA
authors.lastname: High
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D004285:Dogs; D005164:Factor IX / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D002836:Hemophilia B / Q000276:immunology / Q000628:therapy*; D006801:Humans; D007273:Injections, Intramuscular; D008297:Male; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: D004279:DNA, Viral; D005164:Factor IX