Development of gene therapy using a novel domestic viral vector.

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Hematopoietic stem cells (HSCs) are a promising target for gene therapy, however, the low efficiencies of gene transfer using currently available vectors face practical limitations. We have recently developed a novel and efficient gene transfer agent, namely recombinant Sendai virus (SeV). We characterized SeV-mediated gene transfer to human cord blood (CB) HSCs and primitive progenitor cells (PPC) using the jelly fish green fluorescent protein (GFP) gene. Even at a relatively low titer, SeV achieved highly efficient GFP expression in GB CD34^+ cells, as well as more immature CB progenitor cells, CD34^+AC133^+ and GD34^+CD38^-cells, without cytokines prestimulation, that was a clear contrast to the features of gene, transfer using retroviruses. SeV-mediated gene transfer was not seriously affected by the cell cycle status : GFP was expressed in significant percentage of the G_0 cells. In vitro cell differentiation studies revealed that gene transfer occurred in progenitor cells of all lineages (GM-CFU, BFU-E, Mix-CFUand Total). These findings showed that SeV could prove to be a promising vector for efficient gene transfer to CB HSCs, while preserving their ability to reconstitute the entire hematopoietic series. in vivo gene transfer experiments using SeV with temperature-sensitive mutation are now underway.