Lentiviral Vectors for Sustained Transgene Expression in Human Bone Marrow–Derived Stromal Cells

Abstract

Bone marrow-derived mesenchymal stromal cells (MSCs) have attracted attention as potential platforms for the systemic delivery of therapeutic proteins in vivo following gene transfer using oncogenic retroviruses. However, the major limitations of this strategy include low levels of gene transfer and a general lack of long-term transgene expression. We have investigated the expression of several transgenes in MSCs following HIV-1 lentiviral vector-mediated gene transfer. Vectors containing a variety of strong promoters driving enhanced green fluorescence protein (EGFP) and coral (Discosoma sp.)-derived red fluorescent protein (DsRed) reporter genes pseudotyped with the vesicular stomatitis virus-G (VSV-G) glycoprotein were able to transduce cultured MSCs with high efficiency. Transduction efficiencies and transgene expression levels in MSCs were found to be higher with lentiviral vectors than with a vector based on the murine stem cell virus pseudotyped with VSV-G. Transgene expression was maintained in culture for at least 5 months. HIV-1-based lentiviral vectors were able to transduce clonogenic mesenchymal progenitor cells, which were capable of maintaining transgene expression by their MSC progeny, over several cell divisions and during differentiation into adipocytes, indicating that terminal adipocyte cell differentiation was unaffected by lentivirus-mediated reporter gene transfer. Collectively these results suggest that lentivirus-mediated gene transfer strategies provide an efficient tool for ex vivo modification of MSCs that does not interfere with differentiation.

High-Efficiency Transduction of Fibroblasts and Mesenchymal Stem Cells by Tyrosine-Mutant AAV2 Vectors for Their Potential Use in Cellular Therapy

“…Thus, the use of viral vectors for MSC gene transfer has gained prominence, with retroviral and lentiviral vectors being the most rigorously pursued (Chuah et al, 2000;Zhang et al, 2002;Shakhbazau et al, 2008;Barzilay et al, 2009;Uchibori et al, 2009). Second, mouse fibroblasts have been used to develop induced pluripotent stem (iPS) cells by Yamanaka and colleagues (Takahashi and Yamanaka, 2006).…”

Adeno-associated viral gene transfer of transforming growth factor-β1 to human mesenchymal stem cells improves cartilage repair

“…21 Efficient and sustained gene delivery to mesenchymal stem cells with adenovirus, lentivirus and retrovirus have been described. 25 Although viral vectors are effective agents for gene transfer, there are significant safety concerns regarding the use of adenoviruses and retroviruses for clinical gene therapy. Adenovirus is known to induce immunological reactions that can be severe, and even fatal.…”

HIV-1-based defective lentiviral vectors efficiently transduce human monocytes-derived macrophages and suppress replication of wild-type HIV-1

“…Among the cell types which are susceptible to HIV-1-based vectors are hematopoietic stem cells [1][2][3][4][5][6][7] and monocytes/macrophages [8][9][10][11][12][13] which represent important targets for human gene therapy [14][15][16].…”