Efficient regeneration of tumor-specific cytotoxic lymphocytes using the iPSC technology and genome editing

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K15215
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry
• Research Institution: Shiga University of Medical Science
• Principal Investigator: AGATA YASUTOSHI 滋賀医科大学, 医学部, 教授 (60263141)
• Research Collaborator: Kawamoto Hiroshi
• Project Period (FY): 2016-04-01 – 2019-03-31
• 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）

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.

Free Research Field

分子生物学、免疫学

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。