title: Persistent expression of factor VIII in vivo following nonprimate lentiviral gene transfer.
abstract: Hemophilia A is a clinically important coagulation disorder caused by the lack or abnormality of plasma coagulation factor VIII (FVIII). Gene transfer of the FVIII cDNA to hepatocytes using lentiviral vectors is a potential therapeutic approach. We investigated the efficacy of feline immunodeficiency virus (FIV)-based vectors in targeting hepatocytes and correcting FVIII deficiency in a hemophilia A mouse model. Several viral envelope glycoproteins were screened for efficient FIV vector pseudotyping and hepatocyte transduction. The GP64 glycoprotein from baculovirus Autographa californica multinuclear polyhedrosis virus pseudo-typed FIV efficiently and showed excellent hepatocyte tropism. The GP64-pseudotyped vector was stable in the presence of human or mouse complement. Inclusion of a hybrid liver-specific promoter (murine albumin enhancer/human alpha1-antitrypsin promoter) further enhanced transgene expression in hepatocytes. We generated a GP64-pseudotyped FIV vector encoding the B domain-deleted human FVIII coding region driven by the liver-specific promoter, with 2 beneficial point mutations in the A1 domain. Intravenous vector administration conferred sustained FVIII expression in hemophilia A mice for several months without the generation of anti-human FVIII antibodies and resulted in partial phenotypic correction. These findings demonstrate the utility of GP64-pseudotyped FIV lentiviral vectors for targeting hepatocytes to correct disorders associated with deficiencies of secreted proteins.
authors.affiliation: 240G EMRB, Department of Pediatrics, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Yubin
authors.initials: Y
authors.lastname: Kang
authors.forename: Litao
authors.initials: L
authors.lastname: Xie
authors.forename: Diane Thi
authors.initials: DT
authors.lastname: Tran
authors.forename: Colleen S
authors.initials: CS
authors.lastname: Stein
authors.forename: Melissa
authors.initials: M
authors.lastname: Hickey
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
authors.forename: Paul B
authors.initials: PB
authors.lastname: McCray
mesh_terms: D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D004195:Disease Models, Animal; D005169:Factor VIII / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000097:blood / Q000235:genetics / Q000627:therapeutic use*; D006467:Hemophilia A / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D051379:Mice; D008810:Mice, Inbred C57BL; D014018:Tissue Distribution
chemical_list: D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D005169:Factor VIII