Development of antigen specific immune cell therapy based on Crispr/Cas9 genome editing system

• Project/Area Number: 15K14404
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Tumor therapeutics
• Research Institution: Chiba University (2016); The University of Tokyo (2015)
• Principal Investigator: Uematsu Satoshi 千葉大学, 大学院医学研究院, 教授 (50379088)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: CRISPR / Cas9 / ゲノム編集 / がん抗原 / 細胞傷害性T細胞 / Crispr / CRISPR/CAS9 / がん抗原特異的CTL / 抗原特異的な癌免疫細胞療法

Outline of Final Research Achievements

We generated T cell receptors-deficient CD8+ T cell delivering modified spCas9 recombinant protein/gRNA complex targeting on T cell receptors by electroporation. We further insert hgp100 specific T cell receptor genes to T cell receptors-deficient CD8+ T cells and generated hgp100-specific CD8+ T cells. These hgp100-specific CD8+ T cells showed good cytotoxic activity against malignant melanoma. We successfully generated the method to delete TCR genes by CRISPR/Cas9 genome editing system and could make antigen-specific cytotoxic T cells by delivering tumor antigen-specific TCR genes to TCR-deficient cells.

Research Products

• [Int'l Joint Research] パスツール研究所(フランス)
• [Int'l Joint Research] ハーバード大学(米国)
• [Journal Article] G-CSF-induced sympathetic tone provokes fever and primes antimobilizing functions of neutrophils via PGE2. (2017)
  • Author(s): Kawano Y, Fukui C, Shinohara M, Wakahashi K, Ishii S, Suzuki T, Sato M, Asada N, Kawano H, Minagawa K, Sada A, Furuyashiki T, Uematsu S, Akira S, Uede T, Narumiya S, Matsui T, Katayama Y.
  • Journal Title: Blood Volume: 129 Issue: 5 Pages: 587-597
  • DOI: 10.1182/blood-2016-07-725754
  • Peer Reviewed / Open Access
• [Journal Article] Fine Particles Induce Alveolar Macrophage Death and Interleukin-1α Release to Promote Inducible Bronchus-Associated Lymphoid Tissue Formation. (2016)
  • Author(s): Kuroda E, Ozasa K, Temizoz B, Ohata K, Koo CX, Kanuma T, Kusakabe T, Kobari S, Horie M, Morimoto Y, Nakajima S, Kabashima K, Ziegler SF, Iwakura Y, Ise W, Kurosaki T, Nagatake T, Kunisawa J, Takemura N, Uematsu S, Hayashi M, Aoshi T, Kobiyama K, Coban C, Ishii KJ.
  • Journal Title: Immunity. Volume: 45(6) Issue: 6 Pages: 1299-1310
  • DOI: 10.1016/j.immuni.2016.11.010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] p53 is a Host Cell Regulator during Herpes Simplex Encephalitis. (2016)
  • Author(s): Maruzuru Y, Koyanagi N, Takemura N, Uematsu S, Matsubara D, Suzuki Y, Arii J, Kato A, Kawaguchi Y.
  • Journal Title: J Virol. Volume: 90(15) Issue: 15 Pages: 6738-6745
  • DOI: 10.1128/jvi.00846-16
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Epithelial glycosylation in gut homeostasis and inflammation. (2016)
  • Author(s): Goto, Y., Uematsu, S. and Kiyono, H.
  • Journal Title: Nature Immunology Volume: 17 Issue: 11 Pages: 1244-1251
  • DOI: 10.1038/ni.3587
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Isolation and Fuctional Analysis of Lamina PropriaDendritic Cells from the Mouse Small Intestine. (2016)
  • Author(s): Takemura N, Uematsu S.
  • Journal Title: Methods Mol Biol. Volume: 1422 Pages: 181-188
  • DOI: 10.1007/978-1-4939-3603-8_17
  • ISBN: 9781493936014, 9781493936038
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Toll様受容体を標的とした急性放射線性消化管症候群の新たな予防戦略. (2016)
  • Author(s): 武村 直紀, 植松 智
  • Journal Title: 臨床免疫 臨床免疫・アレルギー科 Volume: 66 Pages: 267-273
  • NAID: 40020953204
• [Journal Article] 放射線誘導性細胞死が引き起こす臓器障害に対する自然免疫学的治療戦略. (2016)
  • Author(s): 武村 直紀, 植松 智
  • Journal Title: 実験医学増刊号 Volume: 34 Pages: 110-115
• [Journal Article] Direct Melanoma Cell Contact Induces Stromal Cell Autocrine Prostaglandin E2-EP4 Receptor Signaling that Drives Tumor Growth, Angiogenesis and Metastasis. (2015)
  • Author(s): Inada M, Takita M, Yokoyama S, Watanabe K, Tominari T, Matsumoto C, Hirata M, Maru Y, Maruyama T, Sugimoto Y, Narumiya S, Uematsu S, Akira S, Murphy G, Nagase H, Miyaura C.
  • Journal Title: J Biol Chem. Volume: 290 Issue: 50 Pages: 29781-93
  • DOI: 10.1074/jbc.m115.669481
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Roles of mPGES-1, an inducible prostaglandin E synthase, in enhancement of LPS-induced lymphangiogenesis in a mouse peritonitis model. (2015)
  • Author(s): Matsuda H, Hosono K, Tsuru S, Kurashige C, Sekiguchi K, Akira S, Uematsu S, Okamoto H, Majima M.
  • Journal Title: Life Sci. Volume: 142 Pages: 1-7
  • DOI: 10.1016/j.lfs.2015.10.008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Soluble flagellin co-immunization attenuates Th1 priming to Salmonella and clearance by modulating dendritic cell activation and cytokine production. (2015)
  • Author(s): Flores-Langarica A, Bobat S, Marshall JL, Yam-Puc JC, Cook CN, Serre K, Kingsley RA, Flores-Romo L, Uematsu S, Akira S, Henderson IR, Toellner KM, Cunningham AF.
  • Journal Title: Eur J Immunol. Volume: 45 Issue: 8 Pages: 2299-311
  • DOI: 10.1002/eji.201545564
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 5-azacytidine-induced protein 2 (AZI2) regulates bone mass by fine-tuning osteoclast survival. (2015)
  • Author(s): Maruyama K, Fukasaka M, Uematsu S, Takeuchi O, Kondo T, Saitoh T, Martino M, Akira S.
  • Journal Title: J Biol Chem Volume: 290 Issue: 15 Pages: 9377-9386
  • DOI: 10.1074/jbc.m114.631374
  • Peer Reviewed / Open Access
• [Presentation] 粘膜免疫応答を誘導する新規アジュバントの開発 (2017)
  • Author(s): 植松智
  • Organizer: 第10回次世代アジュバント研究会
  • Place of Presentation: 大阪府豊中市新千里東町1-4-2 千里ライフサイエンスセンター
  • Invited
• [Presentation] 慢性放射線腸障害における好酸球の必須の役割 (2016)
  • Author(s): 植松智
  • Organizer: 第40回日本リンパ学会総会 シンポジウム発表
  • Place of Presentation: 東京都文京区本郷7-3-1 伊藤国際学術研究センター
  • Invited
• [Presentation] 急性、慢性放射線腸障害における自然免疫の役割 (2016)
  • Author(s): 植松智
  • Organizer: 第53回日本消化器免疫学会総会 特別講演
  • Place of Presentation: 大阪市中央区城見1-4-1 ホテルニューオータニ大阪
  • Invited
• [Presentation] 急性、慢性放射線腸障害における ダイイングコードの解明 (2016)
  • Author(s): 植松智
  • Organizer: 第25回日本Cell Death学会学術集会
  • Place of Presentation: 東京都品川区 品川区立総合区民会館 きゅりあん
  • Invited
• [Presentation] Acute and chronic radiation injury in small intestine. (2015)
  • Author(s): Satoshi Uematsu.
  • Organizer: 第４４回日本免疫学会学術集会.シンポジウム.
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2015-11-18
  • Invited
• [Presentation] Resident myofibroblasts and eosinophils create a mutual activation loop for submucosal fibrosis in small intestine after abdominal irradiation. (2015)
  • Author(s): Satoshi Uematsu.
  • Organizer: Toll 2015
  • Place of Presentation: マルベラ、スペイン
  • Year and Date: 2015-09-30
  • Int'l Joint Research
• [Presentation] 消化管自然免疫からみた食物アレルギーの成立機構. (2015)
  • Author(s): 植松智
  • Organizer: 第４６回日本職業・環境アレルギー学会総会・学術大会シンポジウム
  • Place of Presentation: 東京大学伊藤国際学術研究センター（東京都文京区）
  • Year and Date: 2015-07-03
  • Invited
• [Book] 次世代アジュバント開発のためのメカニズム解明と安全性評価 (2017)
  • Author(s): 武村 直紀、植松 智
  • Total Pages: 9
  • Publisher: シーエムシー出版
• [Book] 炎症と免疫「放射線誘導性腸管上皮傷害におけるTLR3の役割」 (2016)
  • Author(s): 武村 直紀, 植松 智.
  • Total Pages: 7
  • Publisher: 先端医学社
• [Book] 臨床免疫・アレルギー科「小腸粘膜固有層の自然免疫細胞が担う腸管免疫応答. 」 (2015)
  • Author(s): 武村 直紀, 植松 智.
  • Total Pages: 8
  • Publisher: 科学評論社
• [Book] 臨床免疫・アレルギー科「放射線誘導性小腸上皮傷害におけるTLR3の役割.」 (2015)
  • Author(s): 武村 直紀, 植松 智.
  • Total Pages: 8
  • Publisher: 科学評論社
• [Book] 医学のあゆみ「Toll様受容体と急性放射線性消化管症候群.」 (2015)
  • Author(s): 武村 直紀, 植松 智.
  • Total Pages: 7
  • Publisher: 医歯薬出版株式会社
• [Remarks] 千葉大学大学院医学研究院・医学部粘膜免疫学
  • URL: http://www.m.chiba-u.ac.jp/class/mucosa/
• [Remarks] 東京大学医科学研究所 国際粘膜ワクチン開発研究センター 自然免疫制御分野
  • URL: http://www.ims.u-tokyo.ac.jp/shizenmenekiseigyo/
• [Patent(Industrial Property Rights)] ワクチン用アジュバント、ワクチン組成物、及び免疫誘導方法 (2016)
  • Inventor(s): 植松智、武村直紀
  • Industrial Property Rights Holder: 植松智、武村直紀
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-06-10
  • Overseas
• [Patent(Industrial Property Rights)] ワクチン用アジュバント、ワクチン組成物、及び免疫誘導方法 (2015)
  • Inventor(s): 植松智 武村直紀
  • Industrial Property Rights Holder: 植松智 武村直紀
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-117871
  • Filing Date: 2015-06-10