title: Sustained high-level expression of human factor IX (hFIX) after liver-targeted delivery of recombinant adeno-associated virus encoding the hFIX gene in rhesus macaques.
abstract: The feasibility, safety, and efficacy of liver-directed gene transfer was evaluated in 5 male macaques (aged 2.5 to 6.5 years) by using a recombinant adeno-associated viral (rAAV) vector (rAAV-2 CAGG-hFIX) that had previously mediated persistent therapeutic expression of human factor IX (hFIX; 6%-10% of physiologic levels) in murine models. A dose of 4 x 10(12) vector genomes (vgs)/kg of body weight was administered through the hepatic artery or portal vein. Persistence of the rAAV vgs as circular monomers and dimers and high-molecular-weight concatamers was documented in liver tissue by Southern blot analysis for periods of up to 1 year. Vector particles were present in plasma, urine, or saliva for several days after infusion (as shown by polymerase chain reaction analysis), and the vgs were detected in spleen tissue at low copy numbers. An enzyme-linked immunosorption assay capable of detecting between 1% and 25% of normal levels of hFIX in rhesus plasma was developed by using hyperimmune serum from a rhesus monkey that had received an adenoviral vector encoding hFIX. Two macaques having 3 and 40 rAAV genome equivalents/cell, respectively, in liver tissue had 4% and 8% of normal physiologic plasma levels of hFIX, respectively. A level of hFIX that was 3% of normal levels was transiently detected in one other macaque, which had a genome copy number of 25 before abrogation by a neutralizing antibody (inhibitor) to hFIX. This nonhuman-primate model will be useful in further evaluation and development of rAAV vectors for gene therapy of hemophilia B.
authors.affiliation: Department of Haematology, University College London, United Kingdom.
authors.forename: Amit C
authors.initials: AC
authors.lastname: Nathwani
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Davidoff
authors.forename: Hideki
authors.initials: H
authors.lastname: Hanawa
authors.forename: Yunyu
authors.initials: Y
authors.lastname: Hu
authors.forename: Fredric A
authors.initials: FA
authors.lastname: Hoffer
authors.forename: Alexander
authors.initials: A
authors.lastname: Nikanorov
authors.forename: Clive
authors.initials: C
authors.lastname: Slaughter
authors.forename: Catherine Y C
authors.initials: CY
authors.lastname: Ng
authors.forename: Junfang
authors.initials: J
authors.lastname: Zhou
authors.forename: Jay N
authors.initials: JN
authors.lastname: Lozier
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Mandrell
authors.forename: Elio F
authors.initials: EF
authors.lastname: Vanin
authors.forename: Arthur W
authors.initials: AW
authors.lastname: Nienhuis
mesh_terms: D000818:Animals; D000229:Dependovirus; D004797:Enzyme-Linked Immunosorbent Assay; D005164:Factor IX / Q000096:biosynthesis / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008253:Macaca mulatta; D008297:Male; D011995:Recombination, Genetic; D014161:Transduction, Genetic
chemical_list: D005164:Factor IX