Efficient lentiviral transduction of liver requires cell cycling in vivo

Abstract

Human-immunodeficiency-virus (HIV)-based lentiviral vectors are a promising tool for in vivo gene therapy. Unlike Moloney-murine-leukaemia-based retroviruses (MLV), lentiviruses are believed to stably transduce quiescent (non-cycling) cells in various organs. No previous studies, however, have directly established the cell-cycle status of any transduced cell type at the time of vector administration in vivo. In vitro studies using wild-type HIV or HIV-based vectors have shown that, in some cases, cell-cycle activation is required for infection, even though cellular mitosis is not an absolute requirement for integration. Even if the block in reverse transcription is overcome in quiescent T cells, productive infection by HIV cannot be rescued in the absence of cell-cycle activation. The potential use of these vectors for gene therapy prompted our study, which establishes a cell-cycle requirement for efficient transduction of hepatocytes in vivo.

A lentiviral vector expressing a fusogenic glycoprotein for cancer gene therapy

“…8 Lentiviral vectors, in contrast, are capable of infecting both dividing and nondividing cells, [10][11][12][13] although a recent report has suggested a cell cycle requirement for transduction of hepatocytes in vivo. 31 We confirmed that the Lenti-GALV, but not the lower titre C-type, stocks could infect both dividing and nondividing human tumour cells in vitro. This means that, although alternative methods of targeting of these vectors must be found to ensure accuracy of gene delivery/expression, 9 both the real titre (s.f.u./ml), and the effective titre in terms of the the potential pool of target tumour cells which they can productively infect, is likely to be higher for the Lenti-GALV vector than for its C-type counterpart.…”

Jaagsiekte sheep retrovirus pseudotyped lentiviral vector-mediated gene transfer to fetal ovine lung

“…Fetal tracheal occlusion rapidly increases lung epithelial proliferation, 47 while increased cell cycling has been shown to enhance lentiviral transduction in vivo. 48 Clinically, fetal tracheal occlusion following lung vector delivery may be feasible using a reversible balloon tracheal occlusion strategy like that used to treat human fetuses with severe congenital diaphragmatic hernia. 49 Development of a lung-targeting viral vector (that is JSRV-LV) was a third strategy we employed to improve transduction efficiency in fetal sheep.…”

Long-term transgene expression by administration of a lentivirus-based vector to the fetal circulation of immuno-competent mice

“…Previously, an HIV-1-derived vector at higher infectious vector titres (2 Â 10 8 TU) showed lowlevel transduction (2.2%) of the liver of C57BL/6-SCID mice. 36 These levels were increased to 60% of hepatocytes by partial hepatectomy and a similar vector dose resulted in 10% hepatocyte expression of GFP in immunocompetent C57/Bl6 mice after portal vein infusion. 21 Vector dose is an important consideration for in utero gene delivery since potential hepatotoxicity could compromise fetal survival.…”