Identification and analyses of colitis-associated gene segment using colitis-model mice by CRISPR/Cas9 system.

• Project/Area Number: 26460972
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Kumamoto University
• Principal Investigator: Irie Atsushi 熊本大学, 大学院生命科学研究部(医), 講師 (30250343)
• Co-Investigator(Kenkyū-buntansha): 今村 隆寿 熊本大学, 大学院生命科学研究部(医), 准教授 (20176499) ; 竹田 直樹 熊本大学, 生命資源研究・支援センター, 助教 (90304998)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 潰瘍性大腸炎 / HLA-DR / CRISPR/Cas9システム / トランスジェニックマウス / 疾患モデル動物 / 潰瘍生大腸炎

Outline of Final Research Achievements

The aim of this study is to identify the cause of spontaneous colitis, which homozygotes of HLA-DR4 transgenic mice established by the investigators develop. At the start point, the transgenes were identified to be inserted in the chromosome 3 with the deletion of about 39kb segment. Since only homozygotes develop colitis, either deletion of the 39kb gene segment or higher expression of HLA-DR4 might be involved in the cause of the colitis.
Using CRISPR/Cas9 system, we have successfully established the mice without the 39kb segment, however, those mice did not develop colitis. Therefore, the 39kb segment may have nothing to do with the development of the colitis. On the other hand, the homozygotes of HLA-DR4 transgenic mice with MHC class II transactivator (CIITA) knockout background, which do not express MHC class II molecules including HLA-DR4, did not develop the colitis. Therefore, overexpression of HLA-DR4, not the lack of the 39kb, is involved in the cause of the colitis.

Research Products

• [Journal Article] Enhancement of active MMP release and invasive activity of lymph node metastatic tongue cancer cells by elevated signaling via the TNF-α-TNFR1-NF-κB pathway and a possible involvement of angiopoietin-like 4 in lung metastasis. (2016)
  • Author(s): Tanaka, T., Imamura, T., Yoneda, M., Irie, A., Ogi, H., Nagata, M., Yoshida, R., Fukuma, D., Kawahara, K., Shinohara, M, & Nakayama, H.
  • Journal Title: Int. J. Oncol. Volume: 49 Issue: 4 Pages: 1377-1384
  • DOI: 10.3892/ijo.2016.3653
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] An oncofetal antigen, IMP-3-derived long peptides induce immune responses of both helper T cells and CTLs (2016)
  • Author(s): M. Hirayama, Y. Tomita, A. Yuno, H. Tsukamoto, S. Sejju, Y. Imamura, M. A. Sayem, A. Irie, Y. Yoshitake, D. Fukuma, M. Shinohara, A. Hamada, H. Jono, E. Yuba, K. Kono, K. Yoshida, T. Tsunoda, H. Nakayama, Y. Nishimura
  • Journal Title: OncoImmunology Volume: 5 Issue: 6 Pages: e1123368-e1123368
  • DOI: 10.1080/2162402x.2015.1123368
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of glypican-3-derived long peptides activating both CD8 and CD4 T cells; prolonged overall survival in cancer patients with Th cell response (2016)
  • Author(s): Sayem, M. A. Tomita, Y. Yuno, A. Hirayama, M. Irie, A. Tsukamoto, H. Senju, S. Yuba, E. Yoshikawa, T. Kono, K. Nakatsura, T. Nishimura, Y.
  • Journal Title: Oncoimmunology Volume: 5 Issue: 1 Pages: e1062209-e1062209
  • DOI: 10.1080/2162402x.2015.1062209
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Influence of the C5a-C5a receptor system on breast cancer progression and patient prognosis (2015)
  • Author(s): Imamura, T. Yamamoto-Ibusuki, M. Sueta, A. Kubo, T. Irie, A. Kikuchi, K. Kariu, T. Iwase, H.
  • Journal Title: Breast Cancer Volume: Oct21 Issue: 6 Pages: 1-10
  • DOI: 10.1007/s12282-015-0654-3
  • Peer Reviewed / Open Access
• [Journal Article] Identification of immunogenic LY6K long peptide encompassing both CD4+ and CD8+ T-cell epitopes and eliciting CD4+ T-cell immunity in patients with malignant disease. (2014)
  • Author(s): Tomita, Y., Yuno, A., Tsukamoto, H., Senju, S., Kuroda, Y., Hirayama, M., Imamura, Y., Yatsuda, J., Sayem, M. A., Irie, A., Hamada A., Jono, H., Yoshida, K., Tsunoda, T, Daigo, Y., Kohrogi, H., Yoshitake, Y., Nakamura,Y., Shinohara, M. and Nishimura, Y.
  • Journal Title: OncoImmunology Volume: 3 Issue: 3 Pages: e28100-e28100
  • DOI: 10.4161/onci.28100
  • Peer Reviewed / Open Access
• [Journal Article] Identification of CDCA1-derived long peptides bearing both CD4+ and CD8+ T-cell epitopes: CDCA1-specific CD4+ T-cell immunity in cancer patients (2014)
  • Author(s): Tomita Y, Yuno A, Tsukamoto H, Senju S, Yoshimura S, Osawa R, Kuroda Y, Hirayama M, Irie A, Hamada A, Jono H, Yoshida K, Tsunoda T, Kohrogi H, Yoshitake Y, Nakamura Y, Shinohara M, Nishimura Y
  • Journal Title: Int J Cancer Volume: 134 Issue: 2 Pages: 352-366
  • DOI: 10.1002/ijc.28376
  • Peer Reviewed
• [Journal Article] S-Guanylation proteomics for redox-based mitochondrial signaling. (2014)
  • Author(s): Rahaman MM, Sawa T, Ahtesh am AK, Khan S, Inoue H, Irie A. Fujii S, and Akaike T.
  • Journal Title: Antioxid. Redox Signal. Volume: 20 Issue: 2 Pages: 295-307
  • DOI: 10.1089/ars.2012.4606
  • Peer Reviewed / Open Access
• [Presentation] Endoplasmic reticulum stress causes ulcerative colitis-like severe colitis in the homozygotes of HLA-DR4 transgenic mice. (2016)
  • Author(s): IRIE A, MICHIBATA Y, KUBO T, IMAMURA T, TAKEDA N, ARAKI K, SHIBUYA I, SOGO S, NISHIMURA Y
  • Organizer: 第45回日本免疫学会学術集会
  • Place of Presentation: 宜野湾
  • Year and Date: 2016-12-05
• [Presentation] 大腸上皮細胞の小胞体ストレスがHLA-DR4トランスジェニックマウスのホモ接合体に発症する大腸炎の病因である (2016)
  • Author(s): 入江 厚、今村 隆寿、道端 弥生、久保 多津子、竹田 直樹、澁谷 功、十河 真司、荒木 喜美、西村 泰治
  • Organizer: 第25回日本組織適合性学会大会
  • Place of Presentation: 札幌
  • Year and Date: 2016-10-22
• [Presentation] Aberrant accumulation of HLA-DR and reduced mucin production in colonic epithelial cells of HLA-DR4 transgenic mice and ulcerative colitis patients. (2016)
  • Author(s): Irie A, Imamura T, Michibata Y, Kubo T, Takeda N, Shibuya I, Sogo S, Araki K, Nishimura Y.
  • Organizer: 16th International Congress of Immunology
  • Place of Presentation: Melbourune, Australia
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Presentation] Endoplasmic reticulum stress causes ulcerative colitis-like severe colitis in the homozygotes of HLADR4 transgenic mice. (2015)
  • Author(s): 入江厚、道端弥生、久保多津子、今村隆寿、竹田直樹、荒木喜美、 渋谷功、十河真司、西村泰治
  • Organizer: 第44回日本免疫学会総会
  • Place of Presentation: 札幌
  • Year and Date: 2015-11-28
• [Presentation] 大腸上皮細胞の小胞体ストレスがHLA-DR4トランスジェニックマウスのホモ接合体に発症する大腸炎の病因である (2015)
  • Author(s): 入江厚、道端弥生、久保多津子、今村隆寿、竹田直樹、荒木喜美、 渋谷功、十河真司、西村泰治
  • Organizer: 第24回日本組織適合性学会大会
  • Place of Presentation: 水戸
  • Year and Date: 2015-09-10
• [Presentation] HLA-DR4トランスジェニックマウスのホモ接合体に発症するリンパ組織形成不全と大腸炎の解析 (2015)
  • Author(s): 入江厚、道端弥生、久保多津子、今村隆寿、竹田直樹、荒木喜美、 渋谷功、十河真司、西村泰治
  • Organizer: 25th Kyoto T cell Conference
  • Place of Presentation: 京都
  • Year and Date: 2015-05-15
• [Presentation] Novel HLA-DR4 transgenic mice that develop sever colitis and pneumonia with serious defects in lymphoid organs (2014)
  • Author(s): Irie, A., Michibata, Y., Kubo, T., Imamura, T., Takeda, N., Araki, K., Sasaki, G., Shibuya, I., Sogo, S., and Nishimura, Y.
  • Organizer: 第43回日本免疫学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 2014-12-10 – 2014-12-12
• [Presentation] HLA-DR4トランスジェニックマウスのホモ接合体のリンパ組織形成不全と大腸炎および肺炎の発症 (2014)
  • Author(s): 入江 厚、道端 弥生、久保 多津子、今村 隆寿、矢津田 旬二、竹田 直樹、荒木 喜美、江藤 正俊、澁谷 功、十河 真司、西村 泰治
  • Organizer: 第23回日本組織適合性学会大会
  • Place of Presentation: 長崎
  • Year and Date: 2014-09-13 – 2015-09-15
• [Book] 免疫生物学原著第９版 第６章Ｔ細胞への抗原提示 （翻訳） (2017)
  • Author(s): 入江 厚、西村 靖治
  • Publisher: 南江堂
• [Remarks] 熊本大学大学院生命科学研究部免疫識別学
  • URL: http://www.immgenet.jp/