Elucidation of physiological roles of NLRP3 phosphorylation in the NLRP3 inflammasome formation

• Project/Area Number: 16H05178
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Experimental pathology
• Research Institution: Nippon Medical School (2018-2019); Keio University (2016-2017)
• Principal Investigator: Morita Rimpei 日本医科大学, 大学院医学研究科, 大学院教授 (00362541)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2018: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2017: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000); Fiscal Year 2016: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
• Keywords: 炎症 / マクロファージ / インフラマソーム / プログラム細胞死 / ミトコンドリア / アクチン / 腫瘍 / NLRP3 / タンパク相互作用 / リン酸化修飾 / BTK / 自然免疫 / 免疫学 / タンパク翻訳後修飾 / サイトカイン

Outline of Final Research Achievements

A pro-inflammatory cytokine IL-1b requires caspase-1 activation through NLRP3 inflammasome formation to be secreted extracellularly. We have previously reported that a non-receptor tyrosine kinase BTK is essential for NLRP3 inflammasome activation. At the beginning, in this study we had tried to determine amino acid residues of NLRP3 phosphorylated by BTK and to clarify its physiological significance. However, we met technical difficulties to purify a sufficient amount of phosphorylated NLRP3 for LC/LC-MS analysis and another research group reported the significance of phosphorylated NLRP3. Thus, we made an alternative of identifying a partner of NLRP3 in macrophages. As a result, we have found that an actin-modulating factor gelsolin negatively regulates NLRP3 inflammasome activation as well as apoptosis on the mitochondria.

Academic Significance and Societal Importance of the Research Achievements

本研究によりgelsolinは細胞骨格アクチンを調整するのみならず、炎症性マクロファージのプログラム細胞死（pyroptosisとapoptosis）を抑制し生存を維持するユニークな因子であることが明らかになりつつある。これはいまだ不明である炎症環境下におけるマクロファージの生存維持が炎症反応の経過と組織修復に与える意義の解明につながる。感染症のみならずがんや生活習慣病にも炎症反応は必ず伴いその病態を修飾することから、本成果は多くの疾患の病態解明につながると期待される。

Research Products

• [Int'l Joint Research] 延世大学校医科大学(韓国)
• [Int'l Joint Research] 延世大学校医科大学(韓国)
• [Int'l Joint Research] Yonsei University College of Medicine(韓国)
• [Journal Article] Distribution of dendritic cells in the septate uterus: An immunological perspective. (2020)
  • Author(s): Negishi Y1,2, Kato M2, Ono S2, Kuwabara Y2, Morita R1, Takahashi H1, Takeshita T2.
  • Journal Title: Am J Reprod Immunol. Volume: e13241 Issue: 6
  • DOI: 10.1111/aji.13241
  • Peer Reviewed
• [Journal Article] A combination of check-point blockade and α-galactosylceramide elicits long-lasting suppressive effects on murine hepatoma cell growth in vivo. (2020)
  • Author(s): Ishii K, Shimizu M, Kogo H, Negishi Y, Tamura H, Morita R, Takahashi H.
  • Journal Title: Immunobiology Volume: 225 Pages: 151860-151860
  • Peer Reviewed
• [Journal Article] Multi-dimensional analysis identified rheumatoid arthritis-driving pathway in human T cell. (2019)
  • Author(s): Takeshita M, Suzuki K, Kondo Y, Morita R, Okuzono Y, Koga K, Kassai Y, Gamo K, Takiguchi M, Kurisu R, Mototani H, Ebisuno Y, Yoshimura A, Takeuchi T
  • Journal Title: Ann Rheum Dis. Volume: 78 Issue: 10 Pages: 1346-1356
  • DOI: 10.1136/annrheumdis-2018-214885
  • Peer Reviewed
• [Journal Article] Significant association between joint ultrasonographic parameters and synovial inflammatory factors in rheumatoid arthritis. (2019)
  • Author(s): Kondo Y, Suzuki K, Inoue Y, Sakata K, Takahashi C, Takeshita M, Kassai Y, Miyazaki T, Morita R, Niki Y, Kaneko Y, Yasuoka H, Yamaoka K, Yoshimura A, Takeuchi T.
  • Journal Title: Arthritis Res Ther. Volume: 21 Issue: 1 Pages: 14-14
  • DOI: 10.1186/s13075-018-1802-x
  • Peer Reviewed
• [Journal Article] Multi-omics monitoring of drug response in rheumatoid arthritis in pursuit of molecular remission. (2018)
  • Author(s): Tasaki S, Suzuki K, Kassai Y, Takeshita M, Murota A, Kondo Y, Ando T, Nakayama Y, Okuzono Y, Takiguchi M, Kurisu R, Miyazaki T, Yoshimoto K, Yasuoka H, Yamaoka K, Morita R, Yoshimura A, Toyoshiba H, Takeuchi T.
  • Journal Title: Nat Commun Volume: 9(1) Issue: 1 Pages: 2755-2755
  • DOI: 10.1038/s41467-018-05044-4
  • Peer Reviewed
• [Journal Article] Generation and application of human induced-stem cell memory T cells for adoptive immunotherapy. (2018)
  • Author(s): Taisuke Kondo Yuki Imura Shunsuke Chikuma Sana Hibino Setsuko Omata‐Mise Makoto Ando Takashi Akanuma Mana Iizuka Ryota Sakai Rimpei Morita Akihiko Yoshimura
  • Journal Title: Cancer Science Volume: 109 Issue: 7 Pages: 2130-2140
  • DOI: 10.1111/cas.13648
  • Peer Reviewed / Open Access
• [Journal Article] Notch-mediated conversion of activated T cells into stem cell memory-like T cells for adoptive immunotherapy (2017)
  • Author(s): Taisuke Kondo, Rimpei Morita, Yuumi Okuzono, Hiroko Nakatsukasa, Takashi Sekiya, Shunsuke Chikuma, Takashi Shichita, Mitsuhiro Kanamori, Masato Kubo, Keiko Koga, Takahiro Miyazaki, Yoshiaki Kassai, Akihiko Yohimura
  • Journal Title: Nature Communications Volume: 8 Issue: 1 Pages: 15338-15338
  • DOI: 10.1038/ncomms15338
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Multiomic disease signatures converge to cytotoxic CD8 T cells in primary Sjögren's syndrome. (2017)
  • Author(s): Tasaki S1,2, Suzuki K3, Nishikawa A3, Kassai Y4, Takiguchi M4, Kurisu R4, Okuzono Y4, Miyazaki T4,5, Takeshita M3, Yoshimoto K3, Yasuoka H3, Yamaoka K3, Ikeura K6, Tsunoda K6, Morita R7, Yoshimura A7, Toyoshiba H1, Takeuchi T3.
  • Journal Title: Ann Rheum Dis Volume: 76 Issue: 8 Pages: 1458-1466
  • DOI: 10.1136/annrheumdis-2016-210788
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mafb prevents excess inflammation after ischemic stroke by accelerating clearance of danger signals through MSR1. (2017)
  • Author(s): 1)Shichita T, Ito M, Morita R, Komai K, Noguchi Y, Ooboshi H, Koshida R, Takahashi S, Kodama T, Yoshimura A.
  • Journal Title: Nat Med. Volume: in press Issue: 6 Pages: 723-732
  • DOI: 10.1038/nm.4312
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Enhanced IgG4 production by follicular helper 2 T cells and the involvement of follicular helper 1 T cells in the pathogenesis of IgG4-related disease. (2016)
  • Author(s): Akiyama M, Yasuoka H, Yamaoka K, Suzuki K, Kaneko Y, Kondo H, Kassai Y, Koga K, Miyazaki T, Morita R, Yoshimura A, Takeuchi T.
  • Journal Title: Arthritis Res Ther. Volume: 18 Issue: 1
  • DOI: 10.1186/s13075-016-1064-4
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification of definitive serum biomarkers associated with disease activity in primary Sjogren's syndrome. (2016)
  • Author(s): Nishikawa A, Suzuki K, Kassai Y, Gotou Y, Takiguchi M, Miyazaki T, Yoshimoto K, Yasuoka H, Yamaoka K, Morita R, Yoshimura A, Takeuchi T.
  • Journal Title: Arthritis Res Ther. Volume: 18 Issue: 1 Pages: 106-106
  • DOI: 10.1186/s13075-016-1006-1
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Serum proteomic analysis identifies interleukin 16 as a biomarker for clinical response during early treatment of rheumatoid arthritis. (2016)
  • Author(s): Murota A, Suzuki K, Kassai Y, Miyazaki T, Morita R, Kondo Y, Takeshita M, Niki Y, Yoshimura A, Takeuchi T.
  • Journal Title: Cytokine. Volume: 78 Pages: 87-93
  • DOI: 10.1016/j.cyto.2015.12.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Resolution of elevated circulating regulatory T cells by corticosteroids in patients with IgG4-related dacryoadenitis and sialoadenitis. (2016)
  • Author(s): Akiyama M, Suzuki K, Kassai Y, Miyazaki T, Morita R, Yoshimura A, Takeuchi T.
  • Journal Title: Int J Rheum Dis. Volume: 19 Issue: 4 Pages: 430-432
  • DOI: 10.1111/1756-185x.12725
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Innate-like function of memory Th17 cells for enhancing endotoxin-induced acute lung inflammation through IL-22. (2015)
  • Author(s): Sakaguchi R, Chikuma S, Shichita T, Morita R, Sekiya T, Ouyang W, Ueda T, Seki H, Morisaki H, Yoshimura A
  • Journal Title: Int Immunol. Volume: 28 Issue: 5 Pages: 233-243
  • DOI: 10.1093/intimm/dxv070
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Clathrin heavy chain is an NLRP3 partner for NLRP3 inflammasome activation (2017)
  • Author(s): 森田林平、吉村昭彦
  • Organizer: 第46回日本免疫学会学術集会
• [Presentation] Notch シグナルによる活性化 T 細胞からステムセルメモリー T 細胞への転換と免疫療法への応用 (2017)
  • Author(s): 近藤泰介、森田林平、吉村昭彦
  • Organizer: 第21回日本がん免疫学会総会