| Project/Area Number |
14570752
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Kyushu University |
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Principal Investigator |
KUSUHARA Koichi Kyushu University, Graduate School of Medical Sciences, Associate Prof., 大学院・医学研究院, 助教授 (20243941)
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| Co-Investigator(Kenkyū-buntansha) |
YONEMITSU Yoshikazu Kyushu University, University Hospital, Assistant Prof., 大学病院, 講師 (40315065)
NOMURA Akihiko Kyushu University, Graduate School of Medical Sciences, Instructor, 大学院・医学研究院, 助手 (00325531)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | gene therapy / hematopoietic stem cells / gene transfer / Sendai virus / vector / cord blood |
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| Research Abstract |
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.
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