Highly efficient lentiviral vector-mediated transduction of nondividing, fully reimplantable primary hepatocytes.
Gene therapy is an attractive approach for the treatment of liver disease. We demonstrate that a so-called third-generation human immunodeficiency virus (HIV)-derived vector system can govern the efficient delivery, integration, and stable expression of a transgene into primary human hepatocytes in the complete absence of cell division. We also show that rodent hepatocytes exhibit a significant degree of resistance to HIV vector-mediated transduction, a phenotype that is particularly pronounced in murine hepatocytes and that results from a block in the immediate-early phase of infection. We finally describe a methodology, that allows very high rates of transduction through minimal in vitro manipulation, in which hepatocytes are kept in suspension and reimplanted within a few hours of harvest with a fully preserved engraftment potential. These results have immediate implications for the treatment of liver diseases by the transplantation of genetically modified hepatocytes, an approach that could be applied to a number of hereditary and acquired hepatic disorders.
Nguyen
Tuan Huy
TH
Departments of Genetics and Microbiology, Faculty of Medicine, University of Geneva, Switzerland.
Oberholzer
José
J
Birraux
Jacques
J
Majno
Pietro
P
Morel
Philippe
P
Trono
Didier
D
D000818:Animals; D002455:Cell Division; D002460:Cell Line; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics; D006367:HeLa Cells; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D066298:In Vitro Techniques; D016086:Lentivirus / Q000235:genetics*; D008107:Liver Diseases / Q000628:therapy; D008297:Male; D051379:Mice; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic