| Project/Area Number |
16K10752
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Neurosurgery
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
難波 宏樹 浜松医科大学, 医学部, 教授 (60198405)
山崎 友裕 浜松医科大学, 医学部附属病院, 助教 (40781050)
小泉 慎一郎 浜松医科大学, 医学部附属病院, 助教 (10456577)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 自殺遺伝子幹細胞療法 / HSVｔｋ/GCV ｓｙｓｔｅｍ / ＰｉｇｇｙＢａｃ system / non-viral vector / HSVtk / エピソーマルベクター / HSVtk/GCV system / 脳腫瘍学 / 自殺遺伝子療法 |
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| Outline of Final Research Achievements |
We tried to transfect the episomal vector with HSVtk gene to stem cells, aiming at the development of stem cell therapy for glioblastoma by non-viral gene transfer method. However, that did not go well because the size of the episomal vector was too large, leading to the low transfer efficiency. So we have devised a non-viral method using another system in which the transgenes are integrated into the host gene. The immortalizing gene is co-transfected in order to achieve immortalization of HSVtk-expressiong stem cells. The mesenchymal stem cells as the vehicle are provided by the company that performs cell preparation based on GMP standards. We have already confirmed the sufficient migratory capacity and cell proliferation ability of the stem cells. We have also prepared the plasmids of several patterns with the combination of the HSVtk, hygromycin, GFP and immotalizing gene so as not to be overlarge size. We are under plasmid transfection by electroporation and lipofection methods.
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| Academic Significance and Societal Importance of the Research Achievements |
ウイルスベクターを用いた遺伝子導入細胞は製剤化を考慮した場合、臨床使用が可能な安全で純度の高い製剤を作成可能な施設は非常に限定されている。そこで非ウイルス的HSVtk遺伝子導入幹細胞株の樹立を目指し本研究を着想した。エピソーマルベクターは自己増殖可能なプラスミドベクターであり、本プラスミドを用いた新規治療法の実現が期待されたがベクターサイズの壁に阻まれ断念した。しかし、引き続き考案したpiggyBacシステムは同様に非ウイルス的遺伝子導入法であるだけでなく、宿主遺伝子に安定的に目的遺伝子が挿入される手法であり、本法は有効性が高く、安全で高品質な細胞製剤の低コスト新規作成法となる。
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