Mechanisms of the development of colitis in T cell-specific Rap1-deficient mice

• Project/Area Number: 19K07612
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49070:Immunology-related
• Research Institution: The University of Tokyo (2022); Kitasato University (2019-2021)
• Principal Investigator: Koko Katagiri 東京大学, 大学院総合文化研究科, 特任研究員 (00322157)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: colitis / Rap1 / TCR / regulatory T cells / microRNA / lymphopenia / proliferation / IBD / T cells / Treg / Actin foci / alpha4beta7 / microbiota / miRNA / Th17 / helper T cells / integrin

Outline of Research at the Start

独自に開発した大腸炎・がん自然発症モデルマウスを用いて、１）T細胞におけるRap1欠損が大腸炎発症につながる新規メカニズムを明らかにする。２）活性型alpha4beta7を特異的に認識するモノクローナル抗体を作成し、その大腸炎治療効果を検討する。また、Rap1がalpha4beta7の構造変化を抑制する分子機講を解明する。３）大腸炎から大腸がんへの進行に関与する因子をプロテオミクス解析及びメタボローム解析により特定する。さらに、４）モデルマウスを用いて明らかになった大腸炎・がん原因分子に関して、ヒトIBD患者T細胞を用いた解析を行い、IBDの原因因子の可能性を検討する。

Outline of Final Research Achievements

T-cell-specific Rap1 deletion causes spontaneous colitis in mice. In the large intestinal lamina propria of these mice, RORgammat+Treg cells were scarcely induced by 4 weeks of age. The expression of CTLA-4 on Rap1-deficient Treg cells was impaired. Rap1-deficient T cells exhibited the defective nuclear translocation of NFAT and actin foci in response to TCR engagement, which is critical for Treg-mediated control of colitogenic Th17 responses. Integrin activation is associated with conformational regulation. We generated two kinds of mAbs that recognized Mn2+-dependent epitopes of β7. Using these mAbs, we found that the conversion of Rap1-GDP to GTP exerts two distinct effects stepwise on the conformation of α4β7. Colitogenic CD4+ T cells demonstrated the increased expression of miR-150. MiR-150 silencing completely inhibited the expansion of pathogenic Th17 cells and the development of colitis in Rap1KO mice. MiR-150 is a potential therapeutic target of IBD.

Academic Significance and Societal Importance of the Research Achievements

Rap1シグナルがTCR distal signalに重要な役割を果たすこと及びRap1-GDPがインテグリンalpah4beta7の活性型構造を抑制していることを初めて明らかにした。また、miR-150の阻害は、リンパ節におけるLIP(lymphopenia-induced proliferation)を抑制し、大腸炎の発症を抑制することから、miR-150阻害剤はIBDなどの自己免疫疾患治療薬として有望であることを見出した。

Research Products

• [Journal Article] Deletion of miR-150 Prevents Spontaneous T Cell Proliferation and the Development of Colitis (2023)
  • Author(s): Ishihara S*, Sato M*, Miyazaki H, Saito H, Sato T, Fujikado N, Sawai S, Kotani A, Katagiri K.
  • Journal Title: Gastro Hep Advances Volume: 2 Issue: 4 Pages: 487-496
  • DOI: 10.1016/j.gastha.2023.01.021
  • Peer Reviewed / Open Access
• [Journal Article] Rap1 prevents colitogenic Th17 cell expansion and facilitates Treg cell differentiation and distal TCR signaling (2022)
  • Author(s): Ishihara Sayaka、Sato Tsuyoshi、Fujikado Noriyuki、Miyazaki Haruka、Yoshimoto Takayuki、Yamamoto Hiromitsu、Fukuda Shinji、Katagiri Koko
  • Journal Title: Communications Biology Volume: 5 Issue: 1 Pages: 206-223
  • DOI: 10.1038/s42003-022-03129-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rap1 is essential for B-cell locomotion, germinal center formation and normal B-1a cell population. (2021)
  • Author(s): Ishihara S, Sato T, Sugioka R, Miwa R, Saito H, Sato R, Fukuyama H, Nakajima A, Sawai S, Kotani A, Katagiri K.
  • Journal Title: Front Immunol. Volume: 12 Pages: 624419-624419
  • DOI: 10.3389/fimmu.2021.624419
  • Peer Reviewed / Open Access
• [Journal Article] Comparative mapping of crawling-cell morphodynamics in deep learning-based feature space (2021)
  • Author(s): Imoto, D., Saito, N., Nakajima, A., Honda, G., Ishida, M., Sugita, T., Ishihara, S., Katagiri, K., Okimura, C., Iwadate, Y. and Sawai, S.
  • Journal Title: PLoS Computational Biology Volume: 17(8) Issue: 8 Pages: 1009237-1009267
  • DOI: 10.1371/journal.pcbi.1009237
  • Peer Reviewed / Open Access
• [Journal Article] Phosphatidic acid-dependent localization and basal de-phosphorylation of RA-GEFs regulate lymphocyte trafficking (2020)
  • Author(s): Ishihara Sayaka、Sato Tsuyoshi、Du Guangwei、Guardavaccaro Daniele、Nakajima Akihiko、Sawai Satoshi、Kataoka Tohru、Katagiri Koko
  • Journal Title: BMC Biology Volume: 18 Issue: 1 Pages: 75-75
  • DOI: 10.1186/s12915-020-00809-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Overexpression of miR-669m inhibits erythroblast differentiation (2020)
  • Author(s): Kotaki R., et.al. Kawashima M, Yamaguchi A, Suzuki N, Koyama-Nasu R, Ogiya D, Okuyama K, Yamamoto Y, Takamatsu M, Kurosaki N, Ando K, Murata A, Ohtsuka M, Nakagawa S, Katagiri K, Kotani A.
  • Journal Title: Sci Rep. Volume: 10 Issue: 1 Pages: 13554-13554
  • DOI: 10.1038/s41598-020-70442-y
  • Peer Reviewed / Open Access
• [Journal Article] Dissection of α 4 β 7 integrin regulation by Rap1 using novel conformation-specific monoclonal anti-β 7 antibodies. (2020)
  • Author(s): Sato T, Ishihara S, Marui R, Takagi J, Katagiri K
  • Journal Title: Sci Rep. Volume: 10 Issue: 1 Pages: 13221-13221
  • DOI: 10.1038/s41598-020-70111-0
  • Peer Reviewed / Open Access
• [Journal Article] Phosphatidic acid regulates subcellular distribution of RA-GEFs critical for chemokine-dependent migration (2020)
  • Author(s): Momoi Yasuyuki、Nishikimi Akihiko、Du Guangwei、Kataoka Tohru、Katagiri Koko
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 524 Issue: 2 Pages: 325-331
  • DOI: 10.1016/j.bbrc.2020.01.080
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] PD-L1/L2 protein levels rapidly increase on monocytes via trogocytosis from tumor cells in classical Hodgkin lymphoma (2020)
  • Author(s): Kawashima M, Carreras J, Higuchi H, Kotaki R, Hoshina T, Okuyama K, Suzuki N, Kakizaki M, Miyatake Y, Ando K, Nakayama M, Umezu S, Horie R, Higuchi Y, Katagiri K, Goyama S, Kitamura T, Chamoto K, Yano S,Nakamura N, and A Kotani
  • Journal Title: Leukemia Volume: - Issue: 9 Pages: 2405-2417
  • DOI: 10.1038/s41375-020-0737-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] HIV Reactivation in Latently Infected Cells With Virological Synapse-Like Cell Contact (2020)
  • Author(s): Okutomi T, Minakawa S, Hirota R, Katagiri K and Morikawa Y.
  • Journal Title: Viruese Volume: - Issue: 4 Pages: 417-417
  • DOI: 10.3390/v12040417
  • Peer Reviewed / Open Access
• [Presentation] Role of Rap1 in preventing colitogenic Th17 cell expansion and in Treg cell differentiation. (2021)
  • Author(s): Haruka Miyazaki, Sayaka Ishihara, Tsuyoshi Sato, Noriyuki Fujikado, Takayuki Yoshimoto, Shinji Fukuda, Koko Katagiri
  • Organizer: 日本免疫学会
• [Presentation] Regulation of TCR signaling by Rap1 (2019)
  • Author(s): Kageyama T, Mamiyoda M, Ishihara S, and Katagiri K.
  • Organizer: 第48回日本免疫学会
• [Presentation] IL-17 plays critical role in the development of spontaneous colitis of T cell-specific Rap1-knockout mice. (2019)
  • Author(s): Hashimoto H, Yoshimoto T, Kodera Y, Ishihara S, and Katagiri K.
  • Organizer: 第48回日本免疫学会
• [Patent(Industrial Property Rights)] 炎症性腸疾患の予防又は治療剤 (2022)
  • Inventor(s): 片桐 晃子
  • Industrial Property Rights Holder: 片桐 晃子
  • Industrial Property Rights Type: 特許
  • Filing Date: 2022
• [Patent(Industrial Property Rights)] 炎症性腸疾患の予防又は治療剤 (2022)
  • Inventor(s): 片桐 晃子
  • Industrial Property Rights Holder: 片桐 晃子
  • Industrial Property Rights Type: 特許
  • Filing Date: 2022