Identification of novel mutations in patients with pyroptosis-associated rare disease and clarification of the pathogenesis

• Project Area: Homeostatic Regulation by Various Types of Cell Death
• Project/Area Number: 17H05515
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Juntendo University
• Principal Investigator: 北浦 次郎 順天堂大学, 医学(系)研究科(研究院), 先任准教授 (30282651)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2018: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2017: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: インフラマソーム / パイロトーシス / 遺伝子 / 細胞・組織 / 免疫学

Outline of Annual Research Achievements

肝臓線維化を主徴とする稀少疾患患者から同定されたＮＬＲＰ１変異が２９３Ｔ細胞を利用した実験系でインフラマソームを異常に強く活性化させ、著しく大量のＩＬ－１ｂやＩＬ－１８を放出させることやパイロトーシスと呼ばれるプログラム細胞死が誘導されることを確認した。この結果と一致して、患者血清中ＩＬ－１８は異常高値を示したが、生体肝移植後には血清中のＩＬ－１８値は著減した。また、生体肝移植を受けた際に摘出された肝臓組織を免疫染色した結果、肝細胞と一致してＩＬ－１８が強く染色された。さらに、患者由来の肝細胞を不死化した結果、恒常的に大量のＩＬ－１８が産生・放出されること、また、この肝細胞株に発現するＮＬＲＰ１をｓｉＲＮＡで特異的にノックダウンするとＩＬ－１８の産生が抑えられることを示した。これらの結果から、ＮＬＲＰ１変異により患者の肝細胞ではＮＬＲＰ１インフラマソームが異常活性化して、ＩＬ－１８の産生や肝細胞のパイロトーシスが亢進し、そのことが肝臓線維化に関与すると考えられた。 ＮＬＲＰ１変異を発現するマウスが作製され、現在、解析中である。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] University Hospital Erlangen/Technische Universität Dresden(Germany)
• [Journal Article] Identification of inhibitory mechanisms in pseudo-allergy involving Mrgprb2/MRGPRX2-mediated mast cell activation (2019)
  • Author(s): Takamori A, Ozawa K, Kaitani A, Ando T, Okamoto Y, Maehara A, Tanabe A, Nagamine M, Yamada H, Uchida S, Uchida K, Isobe M, Hatayama T, Watanabe D, Ando T, Ide T, Matsuzawa M, Maeda K, Nakano N, Tamura N, Ikeda K, Ebihara N, Shimizu T, Ogawa H, Okumura K, Kitaura J
  • Journal Title: Journal of Allergy and Clinical Immunology Volume: 143 Issue: 3 Pages: 1231-1235
  • DOI: 10.1016/j.jaci.2018.10.034
  • Peer Reviewed / Open Access
• [Journal Article] The phytosphingosine-CD300b interaction promotes zymosan-induced, nitric oxide?dependent neutrophil recruitment (2019)
  • Author(s): Takahashi Mariko、Izawa Kumi、Urai Makoto、Yamanishi Yoshinori、Maehara Akie、Isobe Masamichi、Matsukawa Toshihiro、Kaitani Ayako、Takamori Ayako、Uchida Shino、Yamada Hiromichi、Nagamine Masakazu、Ando Tomoaki、Shimizu Toshiaki、Ogawa Hideoki、Okumura Ko、Kinjo Yuki、Kitamura Toshio、Kitaura Jiro
  • Journal Title: Science Signaling Volume: 12 Issue: 564 Pages: 564-564
  • DOI: 10.1126/scisignal.aar5514
  • Peer Reviewed / Open Access
• [Journal Article] IL-31 is crucial for induction of pruritus, but not inflammation, in contact hypersensitivity (2018)
  • Author(s): Takamori Ayako、Nambu Aya、Sato Keiko、Yamaguchi Sachiko、Matsuda Kenshiro、Numata Takafumi、Sugawara Takeru、Yoshizaki Takamichi、Arae Ken、Morita Hideaki、Matsumoto Kenji、Sudo Katsuko、Okumura Ko、Kitaura Jiro、Matsuda Hiroshi、Nakae Susumu
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 6639-6639
  • DOI: 10.1038/s41598-018-25094-4
  • Peer Reviewed / Open Access
• [Journal Article] Leukocyte mono-immunoglobulin-like receptor 8 (LMIR8)/CLM-6 is an FcRγ-coupled receptor selectively expressed in mouse tissue plasmacytoid dendritic cells (2018)
  • Author(s): Kaitani A, Izawa K, Maehara A, Isobe M, Takamori A, Matsukawa T, Takahashi M, Yamanishi Y, Oki T, Yamada H, Nagamine M, Uchida S, Uchida K, Ando T, Maeda K, Nakano N, Shimizu T, Takai T, Ogawa H, Okumura K, Kitamura T, Kitaura J
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 8259-8259
  • DOI: 10.1038/s41598-018-25646-8
  • Peer Reviewed / Open Access
• [Journal Article] The Fab fragment of anti-IgE Cε2 domain prevents allergic reactions through interacting with IgE-FcεRIα complex on rat mast cells. (2018)
  • Author(s): Hirano T, Koyanagi A, Kotoshiba K, Shinkai Y, Kasai M, Ando T, Kaitani A, Okumura K, Kitaura J
  • Journal Title: Sci Rep. Volume: 8 Issue: 1 Pages: 14237-14237
  • DOI: 10.1038/s41598-018-32200-z
  • Peer Reviewed / Open Access
• [Journal Article] Role of the ceramide-CD300f interaction in gram-negative bacterial skin infections. (2018)
  • Author(s): Maehara A, Kaitani A, Izawa K, Shiba E, Nagamine M, Takamori A, Isobe M, Uchida S, Uchida K, Ando T, Keiko M, Nakano N, Voehringer D, Roers A, Shimizu T, Ogawa H, Okumura K, Kitamura T, Kitaura J.
  • Journal Title: J Invest Dermatol. Volume: 138 Issue: 5 Pages: 1221-1224
  • DOI: 10.1016/j.jid.2017.11.025
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The CD300e molecule in mice is an immune-activating receptor. (2018)
  • Author(s): Isobe M, Izawa K, Sugiuchi M, Sakanishi T, Kaitani A, Takamori A, Maehara A, Matsukawa T, Takahashi M, Yamanishi Y, Oki T, Uchida S, Uchida K, Ando T, Maeda K, Nakano N, Yagita H, Takai T, Ogawa H, Okumura K, Kitamura T, Kitaura J.
  • Journal Title: J Biol Chem. Volume: 293 Issue: 10 Pages: 3793-3805
  • DOI: 10.1074/jbc.ra117.000696
  • Peer Reviewed / Open Access
• [Journal Article] High expression of ABCG2 induced by EZH2 disruption has pivotal roles in MDS pathogenesis (2017)
  • Author(s): Kawabata K C、Hayashi Y、Inoue D、Meguro H、Sakurai H、Fukuyama T、Tanaka Y、Asada S、Fukushima T、Nagase R、Takeda R、Harada Y、Kitaura J、Goyama S、Harada H、Aburatani H、Kitamura T
  • Journal Title: Leukemia Volume: 32 Issue: 2 Pages: 419-428
  • DOI: 10.1038/leu.2017.227
  • Peer Reviewed / Open Access
• [Journal Article] Disrupting ceramide-CD300f interaction prevents septic peritonitis by stimulating neutrophil recruitment. (2017)
  • Author(s): Izawa K, Maehara A, Isobe M, Yasuda Y, Urai M, Hoshino Y, Ueno K, Matsukawa T, Takahashi M, Kaitani A, Shiba E, Takamori A, Uchida S, Uchida K, Maeda K, Nakano N, Yamanishi Y, Oki T, Voehringer D, Roers A, Nakae S, Ishikawa J, Kinjo Y, Shimizu T, Ogawa H, Okumura K, Kitamura T, Kitaura J
  • Journal Title: Sci Rep. Volume: 7(1) Issue: 1 Pages: 4298-4298
  • DOI: 10.1038/s41598-017-04647-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] パイロトーシスが関与する稀少疾患の新規遺伝子変異同定と病態解明 (2019)
  • Author(s): 北浦次郎
  • Organizer: 新学術領域研究（平成26-30年度）ダイイングコード 細胞死を起点とする生体制御ネットワークの解明 第4回領域班会議
• [Presentation] Identification and characterization of a novel mutation of NLRP1 in a patient with a rare disease involving severe liver fibrosis. (2018)
  • Author(s): Akie Maehara, Hiroki Yasudo, Atsushi Tanabe, Tomoaki Ando, Kumi Izawa, Ayako Kaitani, Ayako Takamori, Shino Uchida, Keiko Maeda, Nobuhiro Nakano, Ko Okumura, Jiro Kitaura.
  • Organizer: 第2回 Australia-Japan Meeting on Cell Death（新学術領域「ダイイングコード」）
  • Int'l Joint Research
• [Presentation] A novel mutation of NLRP1 is involved in the pathogenesis of a rare disease with severe liver fibrosis. (2018)
  • Author(s): Taiki Ando, Akie Maehara, Tomoaki Ando, Kumi, Izawa, Ayako Kaitani, Atsushi Tanabe, Keiko Maeda, Novuhiro Nakano, Naoto Tamura, Ko Okumura, Jiro Kitaura.
  • Organizer: 第47回日本免疫学会
• [Presentation] パイロトーシスが関与する希少疾患の新規遺伝子変異同定と病態解明 (2018)
  • Author(s): 前原明絵、北浦次郎
  • Organizer: 新学術領域（酵素生物学・ダイイングコード）合同若手会議
• [Presentation] パイロトーシスが関与する稀少疾患の新規遺伝子変異同定と病態解明 (2017)
  • Author(s): 北浦次郎
  • Organizer: 新学術領域研究（平成26-30年度）ダイイングコード 細胞死を起点とする生体制御ネットワークの解明 第3回領域班会議
• [Presentation] Identification and characterization of a novel mutation of NLRP1 in a patient with a rare disease involving severe liver fibrosis (2017)
  • Author(s): Akie Maehara, Atsushi Tanabe, Tomoaki Ando, Kumi Izawa, Ayako Kaitani, Ayako Takamori, Shino Uchida, Keiko Maeda, Nobuhiro Nakano, Ko Okumura, Jiro Kitaura
  • Organizer: 第46回日本免疫学会学術集会