| Project/Area Number |
06559020
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | Okazaki National Research Institutes |
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Principal Investigator |
IKENAKA Kazuhiro Okazaki National Research Institutes, Natl.Inst.For Physiol.Sci., Professor, 生理学研究所, 教授 (00144527)
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| Co-Investigator(Kenkyū-buntansha) |
YOSIMATSU Tadanori Wakunaga Pharmaceutical Co, Ltd.Researcher, バイオ研究所, 副主任
MIYOSHI Kenichi Wakunaga Pharmaceutical Co, Ltd.Director-General, バイオ研究所, 所長
SHIMIZU Keigi Osaka univ.Med.Sch., Associate Professor, 医学部, 助教授 (50162699)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 1995: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1994: ¥3,300,000 (Direct Cost: ¥3,300,000)
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| Keywords | Brain glioma / Gene therapy / HTK gene / Viral particle gene / Liposome / Virus-producer cell / リボソーム法 / グリオーマ / 組み換えレトロウイルス / ヘルペス単純ウィルスチミジンキナーゼ / 腫瘍抑制効果 / by stamder効果 / リポソーム |
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| Research Abstract |
Gene therapy using HTK gene transduction to tumor cells combined with GCV treatment has been applied to human in USA.However, even with a method by direct injection of virus-producer cells, it is thought to be difficult to cure brain glioma completely because of the low titer of retroviral vector, immunological responces for injected cells and highly migratory nature of glioma cells. Therefore we planned to develop a new method in which virus-producer cells are produced from glioma cells by the liposome mediated tranfection with plasmid DNA containing virus particle genes and plasmid DNA containing recombinant retroviral sequence. First, we constructed a vector containing HTK gene deleted with polyA signal. Higher titer of recombinant retrovirus were produced with this vector. In gene therapy using HTK gene, not only HTK transduced tumor cells but untransduced surrounding cells are killed with GCV treatment. This is called bystander effect. We examined efficiency of this effect in our vector system and RSV-M glioma. HTK transduction only into 25% of total glioma cells enabled complete growth suppression of the glioma. For the construction of plasmid containing viral particle gene, gag and pol gene from Psi-2 cells and env gene from PA317 cells were inserted separately to the expression vectors. Recombinant viral particles were not produced by the transduction of these vectors. Now, we are examining the reason for this problem. We also examined the efficient transduction method into murine glioma using liposome.
Efficient transduction was not observed by our experiments using 3 kinds of liposome transfection reagent. Some of them caused toxicity to normal brain tissue.
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