CRISPR-Cas9-Mediated TIM3 Knockout in Human Natural Killer Cells Enhances Growth Inhibitory Effects on Human Glioma Cells.

• Material Type: 博士論文
• Title: CRISPR-Cas9-Mediated TIM3 Knockout in Human Natural Killer Cells Enhances Growth Inhibitory Effects on Human Glioma Cells.
• Author/Editor: Morimoto, Takayuki; Nakazawa, Tsutomu; Matsuda, Ryosuke; Nishimura, Fumihiko; Nakamura, Mitsutoshi; Yamada, Shuichi; Nakagawa, Ichiro; Park, Young-Soo; Tsujimura, Takahiro; Nakase, Hiroyuki
• Author Heading: Morimoto, Takayuki; Nakazawa, Tsutomu; Matsuda, Ryosuke; Nishimura, Fumihiko; Nakamura, Mitsutoshi; Yamada, Shuichi; Nakagawa, Ichiro; Park, Young-Soo; Tsujimura, Takahiro; Nakase, Hiroyuki
• Publication, Distribution, etc.: MDPI
• Publication Date: 2021-03-28
• Publication Date (W3CDTF): 2021-03-28
• Alternative Title: CRISPR-Cas9を用いてTIM3をノックアウトしたヒトNK細胞の膠芽腫細胞株に対する抗腫瘍効果の検討
• Periodical title: International journal of molecular sciences
• No. or year of volume/issue: 22 7
• Volume: 22
• Issue: 7
• Pages: Article No.3489-
• ISSN (Periodical Title): ISSN : 1422-0067
• Degree grantor/type: 奈良県立医科大学
• Date Granted: 2022-03-15
• Dissertation Number: 甲第828号
• Degree Type: 博士（医学）
• Text Language Code: eng
• Subject Heading: TIM3; CRISPR-Cas9; NK cell; glioblastoma
• Target Audience: 一般
• Note (General): type:Thesis; Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor in adults. Natural Killer (NK) cells are potent cytotoxic effector cells against tumor cells inducing GBM cells; therefore, NK cell based- immunotherapy might be a promising target in GBM. T cell immunoglobulin mucin family member 3 (TIM3), a receptor expressed on NK cells, has been suggested as a marker of dysfunctional NK cells. We established TIM3 knockout in NK cells, using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9). Electroporating of TIM3 exon 2- or exon 5-targeting guide RNA- Cas9 protein complexes (RNPs) inhibited TIM3 expression on NK cells with varying efficacy. T7 endonuclease I mutation detection assays showed that both RNPs disrupted the intended genome sites. The expression of other checkpoint receptors, i.e., programmed cell death 1 (PD1), Lymphocyte-activation gene 3 (LAG3), T cell immunoreceptor with Ig and ITIM domains (TIGIT), and TACTILE (CD96) were unchanged on the TIM3 knockout NK cells. Real time cell growth assays revealed that TIM3 knockout enhanced NK cell-mediated growth inhibition of GBM cells. These results demonstrated that TIM3 knockout enhanced human NK cell mediated cytotoxicity on GBM cells. Future, CRISPR-Cas9 mediated TIM3 knockout in NK cells may prove to be a promising immunotherapeutic alternative in patient with GBM.; 博士（医学）・甲第828号・令和4年3月15日; © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).; identifier:International journal of molecular sciences Vol.22 No.7 Article No.3489 (2021 Mar); identifier:14220067; identifier:http://ginmu.naramed-u.ac.jp/dspace/handle/10564/4011; identifier:International journal of molecular sciences, 22(7): Article No.3489
• Source: fulltext; http://ginmu.naramed-u.ac.jp/dspace/bitstream/10564/4011/1/01%e7%94%b2828%e6%9c%ac%e6%96%87%e3%81%ae%e8%a6%81%e6%97%a8.pdf; http://ginmu.naramed-u.ac.jp/dspace/bitstream/10564/4011/2/02%e7%94%b2828%e5%af%a9%e6%9f%bb%e8%a6%81%e6%97%a8.pdf; http://ginmu.naramed-u.ac.jp/dspace/bitstream/10564/4011/3/03%e7%94%b2828%e6%9c%ac%e6%96%87.pdf
• Access Restrictions: インターネット公開
• Related Material (DOI): 10.3390/ijms22073489
• Periodical Title (NCID): AA12038549; https://ci.nii.ac.jp/ncid/AA12038549
• Data Provider (Database): 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）; https://irdb.nii.ac.jp
• Original Data Provider (Database): 奈良県立医科大学 : 奈良県立医科大学機関リポジトリ GINMU