Development of in vivo cytotoxicity assay system using humanized mice maintaining human mature NK cells for long-term

• Project/Area Number: 16K18405
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory animal science
• Research Institution: Central Institute for Experimental Animals
• Principal Investigator: Katano Ikumi 公益財団法人実験動物中央研究所, 実験動物研究部, 研究員 (90442558)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ヒト化マウス / NK細胞 / ヒト免疫 / ADCC / NOGマウス / IL-15 / IL-2 / ヒトNK細胞

Outline of Final Research Achievements

We generated that a NOG-human IL-15 Tg mouse strain was useful for evaluating in vivo antibody-dependent cellular cytotoxicity (ADCC) mediated by human peripheral blood-derived NK cells in the presence of therapeutic antibody.
And, We generated immunoglobulin gamma Fc region receptor (FcRg)-deficient NOG-IL-15 Tg (NOG-FcRg KO IL-15 Tg) mice and investigated whether human NK cell-derived ADCC activity could be distinguished from mouse cell -mediated antibody-dependent cellular phagocytosis (ADCP). The Daudi-transplanted NOG-FcRg KO IL-15 Tg mice were intravenously transferred with in vitro expanded human NK cells and treated with rituximab. Only the group with human NK cells and rituximab treatment showed significant suppression of tumor growth in the kidney, while treatment with rituximab alone had minimum influence.
These data suggest that NOG-FcRg KO, IL-15 mice are more suitable for specifically detecting human NK cell mediated in vivo ADCC activity than NOG-IL-15 Tg mice.

Academic Significance and Societal Importance of the Research Achievements

がんに対する分子標的抗体の創薬研究において、抗体の機能解析は試験管内での検証系が主流である。しかし、この結果は生体での反応を必ずしも反映するものではなく、臨床研究で予期しない副作用が観察されることが多い。従って、ヒトの細胞に対する反応性を生体内で検証し得る動物モデルが求められてきた。従来のモデルでは抗体の細胞傷害機能-ADCC活性-の検証はほぼ不可能であったが、ヒトNK細胞を生着させたNOG-IL-15 Tgマウスで検証が可能となった。さらにNOG-FcRg KO, IL-15 TgマウスではADCC活性の検出感度を飛躍的に高めることができた。この動物モデルは創薬の発展に貢献するものと考える。

Research Products

• [Journal Article] Long-term maintenance of peripheral blood derived human NK cells in a novel human IL-15- transgenic NOG mouse (2017)
  • Author(s): Katano Ikumi、Nishime Chiyoko、Ito Ryoji、Kamisako Tsutomu、Mizusawa Takuma、Ka Yuyo、Ogura Tomoyuki、Suemizu Hiroshi、Kawakami Yutaka、Ito Mamoru、Takahashi Takeshi
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 17230-17230
  • DOI: 10.1038/s41598-017-17442-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A novel xenogeneic graft-versus-host disease model for investigating the pathological role of human CD4+ or CD8+ T cells using immunodeficient NOG mice. (2017)
  • Author(s): Ito R, Katano I, Kawai K, Yagoto M, Takahashi T, Ka Y, Ogura T, Takahashi R, Ito M
  • Journal Title: American Journal of Transplantation Volume: 5 Issue: 5 Pages: 1216-1228
  • DOI: 10.1111/ajt.14116
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Antitumor Effect of Programmed Death-1 (PD-1) Blockade in Humanized the NOG-MHC Double Knockout Mouse. (2017)
  • Author(s): Ashizawa T, Iizuka A, Nonomura C, Kondou R, Maeda C, Miyata H, Sugino T, Mitsuya K, Hayashi N, Nakasu Y, Maruyama K, Yamaguchi K, Katano I, Ito M, Akiyama Y.
  • Journal Title: Clinical Cancer Research Volume: 23(1) Issue: 1 Pages: 149-158
  • DOI: 10.1158/1078-0432.ccr-16-0122
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Generation of a Nonhuman Primate Model of Severe Combined Immunodeficiency Using Highly Efficient Genome Editing. (2016)
  • Author(s): Sato K, Oiwa R, Kumita W, Henry R, Sakuma T, Ito R, Nozu R, Inoue T, Katano I, Sato K, Okahara N, Okahara J, Shimizu Y, Yamamoto M, Hanazawa K, Kawakami T, Kametani Y, Suzuki R, Takahashi T, Weinstein EJ, Yamamoto T, Sakakibara Y, Habu S, Hata J, Okano H, Sasaki E.
  • Journal Title: Cell Stem Cell Volume: 19 Issue: 1 Pages: 127-138
  • DOI: 10.1016/j.stem.2016.06.003
  • Peer Reviewed / Int'l Joint Research
• [Presentation] NOG-FcgR KO-hIL-15 Tg mice provide a highly sensitive assay system for ADCC activity of human NK cells (2019)
  • Author(s): Ikumi Katano, Iyo Ootsuka, Ryoji Ito, Kenji Kawai, Mika Yagoto, Hiroshi Suemizu, Mamoru Ito, Taichi Yamamoto, Takeshi Takahashi
  • Organizer: AACR Annual Meeting 2019
  • Int'l Joint Research
• [Presentation] Specific detection of human NK cell mediated in vivo ADCC in FcgR-deficient NOG-human IL-15 transgenic mice (2018)
  • Author(s): Ikumi Katano, Asami Hanazawa, Ryoji Ito, Mamoru Ito, Takeshi Takahashi
  • Organizer: 第47回 日本免疫学会学術集会
• [Patent(Industrial Property Rights)] 免疫不全マウス (2018)
  • Inventor(s): 高橋武司、片野いくみ
  • Industrial Property Rights Holder: 高橋武司、片野いくみ
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-217229
  • Filing Date: 2018
• [Patent(Industrial Property Rights)] ヒトIL-15分泌免疫不全マウス (2016)
  • Inventor(s): 伊藤守、片野いくみ
  • Industrial Property Rights Holder: 伊藤守、片野いくみ
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-04-20
  • Overseas