title: Preclinical in vivo evaluation of pseudotyped adeno-associated virus vectors for liver gene therapy.
abstract: We report the generation and use of pseudotyped adeno-associated viral (AAV) vectors for the liver-specific expression of human blood coagulation factor IX (hFIX). Therefore, an AAV-2 genome encoding the hfIX gene was cross-packaged into capsids of AAV types 1 to 6 using efficient, large-scale technology for particle production and purification. In immunocompetent mice, the resultant vector particles expressed high hFIX levels ranging from 36% (AAV-4) to more than 2000% of normal (AAV-1, -2, and -6), which would exceed curative levels in patients with hemophilia. Expression was dose- and time-dependent, with AAV-6 directing the fastest and strongest onset of hFIX expression at all doses. Interestingly, systemic administration of 2 x 1012 vector particles of AAV-1, -4, or -6 resulted in hFIX levels similar to those achieved by portal vein delivery. For all other serotypes and particle doses, hepatic vector administration yielded up to 84-fold more hFIX protein than tail vein delivery, corroborated by similarly increased vector DNA copy numbers in the liver, and elicited a reduced immune response against the viral capsids. Finally, neutralization assays showed variable immunologic cross-reactions between most of the AAV serotypes. Our technology and findings should facilitate the development of AAV pseudotype-based gene therapies for hemophilia B and other liver-related diseases.
authors.affiliation: Department of Pediatrics, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Dirk
authors.initials: D
authors.lastname: Grimm
authors.forename: Shangzhen
authors.initials: S
authors.lastname: Zhou
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakai
authors.forename: Clare E
authors.initials: CE
authors.lastname: Thomas
authors.forename: Theresa A
authors.initials: TA
authors.lastname: Storm
authors.forename: Sally
authors.initials: S
authors.lastname: Fuess
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsushita
authors.forename: James
authors.initials: J
authors.lastname: Allen
authors.forename: Richard
authors.initials: R
authors.lastname: Surosky
authors.forename: Michael
authors.initials: M
authors.lastname: Lochrie
authors.forename: Leonard
authors.initials: L
authors.lastname: Meuse
authors.forename: Alan
authors.initials: A
authors.lastname: McClelland
authors.forename: Peter
authors.initials: P
authors.lastname: Colosi
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kay
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000906:Antibodies / Q000097:blood; D002213:Capsid; D006528:Carcinoma, Hepatocellular; D004279:DNA, Viral / Q000493:pharmacokinetics; D005164:Factor IX / Q000235:genetics*; D018628:Gene Dosage; D015967:Gene Expression Regulation, Viral; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000276:immunology / Q000493:pharmacokinetics; D002836:Hemophilia B / Q000628:therapy*; D006801:Humans; D007668:Kidney / Q000166:cytology; D008107:Liver Diseases / Q000628:therapy*; D008113:Liver Neoplasms; D051379:Mice; D008810:Mice, Inbred C57BL; D011401:Promoter Regions, Genetic; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
chemical_list: D000906:Antibodies; D004279:DNA, Viral; D005164:Factor IX