| Project/Area Number |
16K15215
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
General medical chemistry
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
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| Research Collaborator |
Kawamoto Hiroshi
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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 |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | がん免疫 / がん抗原 / Ｔ細胞受容体（TCR） / iPS細胞 / 再生医療 / ゲノム編集 / CRISPR/Cas9 / カセット交換法（RMCE） / Cre/loxP / がん免疫細胞療法 / T-iPS細胞 / TCR-iPS細胞 / Ｔ細胞受容体（TCR）遺伝子 / カセット交換法 / キラーT細胞 |
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| Outline of Final Research Achievements |
In this study, we aimed to efficiently regenerate highly active T cells from iPS cells by knocking in tumor antigen-specific T cell receptor (TCR) genes into the endogenous TCRβ locus. First, we utilized Jurkat cells as a model system and knocked in a drug resistance gene cassette to the endogenous TCRβ locus by genome editing and then successfully exchanged it with the TCR gene by the cassette exchange method using the Cre/loxP system. When we performed the same experiment in iPS cells, we had some problems with cassette exchange, but we achieved reproducible exchange by improvement of the promoter. We are currently attempting to regenerate T cells from iPS cells in which the TCR gene was successfully knocked-in by cassette exchange.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では、がん抗原特異的なTCR遺伝子をiPS細胞の内在性TCRβ遺伝子座へノックインすることに成功した。このiPS細胞から活性の高いT細胞を再生することができれば、カセット交換法により未知変異抗原に反応するTCR遺伝子をノックインしたiPS細胞を次々に作製することも可能となり、細胞製剤として早期の治療応用も期待できる。またレンチウイルスを用いた遺伝子挿入によるがん化のリスクも回避できることから安全性においても有意義である。
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