Efficient gene transduction to cultured hepatocytes by HIV-1 derived lentiviral vector

Abstract

HEPATOCYTE transplantation can be considered a viable and effective replacement therapy for enzymedeficient liver diseases as well as a supportive therapy for many forms of liver failure. However, genetic modifications of hepatocytes in an ex vivo manner may further advance the field of hepatocyte transplantation. A proofof-concept study by Grossman et al demonstrated the clinical feasibility of transplanting ex vivo modified hepatocytes to cure inherited liver disease. Moreover, the allogeneic grafts expressed “protective genes” that could lead to immune tolerance or specific unresponsiveness in the host. One factor critical for the success of ex vivo gene therapy is the development of a method that allows for the longterm, therapeutic expression of the transgene of interest. Among several currently available viral vector systems for use in gene transfer, retroviral vectors based on murine Moloney leukemia virus (MoLV) have been used considerably due to their ability to integrate into the host genome. This would potentially allow for the long-term transgene expression following retroviral integration. However, one major drawback of MoLV is its inability to efficiently transduce genes into quiescent cells, such as primary hepatocytes in culture, due to its need for nuclear membrane dissolution. Unlike MoLV, recently developed lentiviral vectors (LV) derived from HIV-1 have an ability to infect and transduce dividing as well as nondividing cells because of their active transport of preintegration complex through the nuclear pore. We have previously reported that HIV-1 derived LV-mediated gene transduction in the liver in vivo was substantially higher than MoLV vectors. In the present study, we examined the efficiency of LV-mediated transduction in cultured primary mouse hepatocytes.