title: Efficient and Targeted Transduction of Nonhuman Primate Liver With Systemically Delivered Optimized AAV3B Vectors.
abstract: Recombinant adeno-associated virus serotype 3B (rAAV3B) can transduce cultured human liver cancer cells and primary human hepatocytes efficiently. Serine (S)- and threonine (T)-directed capsid modifications further augment its transduction efficiency. Systemically delivered capsid-optimized rAAV3B vectors can specifically target cancer cells in a human liver cancer xenograft model, suggesting their potential use for human liver-directed gene therapy. Here, we compared transduction efficiencies of AAV3B and AAV8 vectors in cultured primary human hepatocytes and cancer cells as well as in human and mouse hepatocytes in a human liver xenograft NSG-PiZ mouse model. We also examined the safety and transduction efficacy of wild-type (WT) and capsid-optimized rAAV3B in the livers of nonhuman primates (NHPs). Intravenously delivered S663V+T492V (ST)-modified self-complementary (sc) AAV3B-EGFP vectors led to liver-targeted robust enhanced green fluorescence protein (EGFP) expression in NHPs without apparent hepatotoxicity. Intravenous injections of both WT and ST-modified rAAV3B.ST-rhCG vectors also generated stable super-physiological levels of rhesus chorionic gonadotropin (rhCG) in NHPs. The vector genome predominantly targeted the liver. Clinical chemistry and histopathology examinations showed no apparent vector-related toxicity. Our studies should be important and informative for clinical development of optimized AAV3B vectors for human liver-directed gene therapy.
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Microbiology and Physiology Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Shaoyong
authors.initials: S
authors.lastname: Li
authors.affiliation: Powell Gene Therapy Center, University of Florida, Gainesville, Florida, USA.|Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida, Gainesville, Florida, USA.
authors.forename: Chen
authors.initials: C
authors.lastname: Ling
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Viral Vector Core, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Zhong
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Microbiology and Physiology Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Mengxin
authors.initials: M
authors.lastname: Li
authors.affiliation: Viral Vector Core, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Su
authors.affiliation: Viral Vector Core, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Ran
authors.initials: R
authors.lastname: He
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Qiushi
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Dale L
authors.initials: DL
authors.lastname: Greiner
authors.affiliation: The Jackson Laboratory Bar Harbor, Bar Harbor, Maine, USA.
authors.forename: Leonard D
authors.initials: LD
authors.lastname: Shultz
authors.affiliation: Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Brehm
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Terence R
authors.initials: TR
authors.lastname: Flotte
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Mueller
authors.affiliation: Powell Gene Therapy Center, University of Florida, Gainesville, Florida, USA.|Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida, Gainesville, Florida, USA.
authors.forename: Arun
authors.initials: A
authors.lastname: Srivastava
authors.affiliation: Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Microbiology and Physiology Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Viral Vector Core, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Guangping
authors.initials: G
authors.lastname: Gao
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000166:cytology; D008113:Liver Neoplasms / Q000235:genetics* / Q000628:therapy; D008253:Macaca mulatta; D051379:Mice; D009368:Neoplasm Transplantation; D014161:Transduction, Genetic
chemical_list: D036022:Capsid Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins