Engulfment of apoptotic cells and asymmetry of plasma membranes

2019 Fiscal Year Annual Research Report

• Project/Area Number: 15H05785
• Research Institution: Osaka University
• Principal Investigator: 長田 重一 大阪大学, 免疫学フロンティア研究センター, 特任教授(常勤) (70114428)
• Project Period (FY): 2015-05-29 – 2020-03-31
• Keywords: アポトーシス / 自然免疫 / リン脂質 / マクロファージ / 貪 食

Outline of Annual Research Achievements

アポトーシスを起こした細胞はフォスファチジルセリン(PtdSer) を “eat me” シグナルとして提示し、マクロファージによって貪食される。私達は、死細胞の貪食に関与する分子 (Tim4, MerTK, MFG-E8)を同定するとともに、PtdSerを細胞膜の内膜と外膜の間でスクランブルさせるスクランブラーゼ (Xkr8およびTMEM16F）、外膜から内膜へ移動させるフリッパーゼ (ATP11Aや11C)を同定した。本研究はこれら分子の作用機序、生理作用を明らかにすることを目的としている。
まず、PtdSerは小胞体 (ER) においても細胞質側に偏在することを見出した。これまで、ERの細胞質側で合成されたPtdSerが一旦、内腔 (lumen) 側に移動、その後細胞膜で再び細胞膜側に転移されると考えられていたが、今回の結果はこの仮説を訂正した。
Xkr8はカスパーゼによって活性化されるとPtdSerを速やかに細胞表面に暴露させる。Xkr8欠損マウスでは、成熟過程で大量に死滅する精原細胞がPtdSerを暴露できず、Sertoli細胞によって貪食されないことが原因で精巣の発達が５週令で停止することを突き止めた。
組織常在マクロファージはPtdSer受容体Tim４を用いて死細胞を認識、チロシンキナーゼ受容体であるであるMerTKを用いて、死細胞を貪食する。アポトーシス細胞はTim4/MerTKを発現している細胞に貪食されるとともに、細胞の増殖も誘導することを見出した。
ATPはその受容体(P2X7)に作用、PtdSerを暴露させる。今回、ERに存在するタンパク質Erosが、P2X7に対するシャペロンとして作用、P2X7を細胞膜に局在させることを見出した。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Remarks

長田重一研究室
http://biochemi.ifrec.osaka-u.ac.jp/

Research Products

• [Int'l Joint Research] Sci and Tech Facilities Council (STFC)(英国)
  • Country Name: UNITED KINGDOM
  • Counterpart Institution: Sci and Tech Facilities Council (STFC)
• [Int'l Joint Research] Cincinnati Children's Hospital Med Cent/St. Jude Children's Research Hospital/University of Vermont(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Cincinnati Children's Hospital Med Cent/St. Jude Children's Research Hospital/University of Vermont
  • # of Other Institutions: 1
• [Int'l Joint Research] Int Cent for Genet Eng & Biotech (ICGEB)(イタリア)
  • Country Name: ITALY
  • Counterpart Institution: Int Cent for Genet Eng & Biotech (ICGEB)
• [Int'l Joint Research] Hadassah-Hebrew Univ Med Center(イスラエル)
  • Country Name: ISRAEL
  • Counterpart Institution: Hadassah-Hebrew Univ Med Center
• [Int'l Joint Research] University Medical Center Groningen(オランダ)
  • Country Name: NETHERLANDS
  • Counterpart Institution: University Medical Center Groningen
• [Int'l Joint Research]
  • # of Other Countries: 3
• [Journal Article] TIM4 on type 1 DCs mediates uptake of tumor-associated antigens and activation of anti tumor responses in lung tumors (2021)
  • Author(s): 1. Caronni, N., Piperno, G., Simoncello, F., Romano, O., Vodret, S., Yanagihashi, Y., Dress, R., Dutertre, C., Bugatti, M., Bourdely, P., Prete, A., Schioppa, T., Mazza, E., Collavin, L., Zacchigna, S., Ostuni, R., Guermonprez, P., Vermi, W., Ginhoux, F., Bicciato, S., Nagata, S., and Benvenuti, F.
  • Journal Title: Nat.Commun Volume: - Pages: in press
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tim4 recognizes carbon nanotubes and mediates phagocytosis leading to granuloma formation (2021)
  • Author(s): Omori Satoshi、Tsugita Misato、Hoshikawa Yasuto、Morita Masanobu、Ito Fumiya、Yamaguchi Shin-Ichiro、Xie Qilin、Noyori Osamu、Yamaguchi Tomoya、Takada Ayato、Saitoh Tatsuya、Toyokuni Shinya、Akiba Hisaya、Nagata Shigekazu、Kinoshita Kengo、Nakayama Masafumi
  • Journal Title: Cell Reports Volume: 34 Pages: 108734～108734
  • DOI: 10.1016/j.celrep.2021.108734
  • Peer Reviewed / Open Access
• [Journal Article] Sensing and clearance of apoptotic cells (2021)
  • Author(s): Nagata Shigekazu、Segawa Katsumori
  • Journal Title: Current Opinion in Immunology Volume: 68 Pages: 1～8
  • DOI: 10.1016/j.coi.2020.07.007
  • Peer Reviewed
• [Journal Article] Transport Cycle of Plasma Membrane Flippase ATP11C by Cryo-EM (2020)
  • Author(s): Nakanishi Hanayo、Nishizawa Tomohiro、Segawa Katsumori、Nureki Osamu、Fujiyoshi Yoshinori、Nagata Shigekazu、Abe Kazuhiro
  • Journal Title: Cell Reports Volume: 32 Pages: 108208～108208
  • DOI: 10.1016/j.celrep.2020.108208
  • Peer Reviewed / Open Access
• [Journal Article] Crystal structure of a human plasma membrane phospholipid flippase (2020)
  • Author(s): Nakanishi Hanayo、Irie Katsumasa、Segawa Katsumori、Hasegawa Kazuya、Fujiyoshi Yoshinori、Nagata Shigekazu、Abe Kazuhiro
  • Journal Title: Journal of Biological Chemistry Volume: 295 Pages: 10180～10194
  • DOI: 10.1074/jbc.RA120.014144
  • Peer Reviewed / Open Access
• [Journal Article] Flippase and scramblase for phosphatidylserine exposure (2020)
  • Author(s): Nagata Shigekazu、Sakuragi Takaharu、Segawa Katsumori
  • Journal Title: Current Opinion in Immunology Volume: 62 Pages: 31～38
  • DOI: 10.1016/j.coi.2019.11.009
  • Peer Reviewed
• [Journal Article] Infertility Caused by Inefficient Apoptotic Germ Cell Clearance in Xkr8-Deficient Male Mice (2019)
  • Author(s): Yamashita Yahiro、Suzuki Chigure、Uchiyama Yasuo、Nagata Shigekazu
  • Journal Title: Molecular and Cellular Biology Volume: 40 Pages: e00402-00419
  • DOI: 10.1128/MCB.00402-19
  • Peer Reviewed
• [Journal Article] Functional Expression of the P2X7 ATP Receptor Requires Eros (2019)
  • Author(s): Ryoden Yuta、Fujii Toshihiro、Segawa Katsumori、Nagata Shigekazu
  • Journal Title: The Journal of Immunology Volume: 204 Pages: 559～568
  • DOI: 10.4049/jimmunol.1900448
  • Peer Reviewed / Open Access
• [Journal Article] Predominant localization of phosphatidylserine at the cytoplasmic leaflet of the ER, and its TMEM16K-dependent redistribution (2019)
  • Author(s): Tsuji Takuma、Cheng Jinglei、Tatematsu Tsuyako、Ebata Aoi、Kamikawa Hiroki、Fujita Akikazu、Gyobu Sayuri、Segawa Katsumori、Arai Hiroyuki、Taguchi Tomohiko、Nagata Shigekazu、Fujimoto Toyoshi
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 116 Pages: 13368～13373
  • DOI: 10.1073/pnas.1822025116
  • Peer Reviewed / Open Access
• [Journal Article] Phosphorylation-mediated activation of mouse Xkr8 scramblase for phosphatidylserine exposure (2019)
  • Author(s): Sakuragi Takaharu、Kosako Hidetaka、Nagata Shigekazu
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 116 Pages: 2907～2912
  • DOI: 10.1073/pnas.1820499116
  • Peer Reviewed / Open Access
• [Journal Article] MERTK tyrosine kinase receptor together with TIM4 phosphatidylserine receptor mediates distinct signal transduction pathways for efferocytosis and cell proliferation (2019)
  • Author(s): Nishi Chihiro、Yanagihashi Yuichi、Segawa Katsumori、Nagata Shigekazu
  • Journal Title: Journal of Biological Chemistry Volume: 294 Pages: 7221～7230
  • DOI: 10.1074/jbc.RA118.006628
  • Peer Reviewed / Open Access
• [Presentation] リン脂質の膜動態とマクロファージ (2020)
  • Author(s): 瀬川 勝盛
  • Organizer: 第93回日本生化学会大会
  • Invited
• [Presentation] アポトーシスと死細胞の貪食 (2019)
  • Author(s): 長田 重一
  • Organizer: 第40回日本炎症・再生医学会
  • Invited
• [Presentation] アポトーシスと死細胞の貪食 (2019)
  • Author(s): 長田 重一
  • Organizer: 第27回日本Cell Death学会
  • Invited
• [Presentation] フリッパーゼとスクランブラーゼ (2019)
  • Author(s): 長田 重一
  • Organizer: 東京大学理学部生物化学科60周年記念シンポジウム
  • Invited
• [Presentation] 死細胞の貪食と細胞膜の非対称性：フリッパーゼとスクランブラーゼ (2019)
  • Author(s): 長田 重一
  • Organizer: 北海道大学遺伝子病制御研究所セミナー
  • Invited
• [Presentation] 細胞の死と死細胞の貪食 (2019)
  • Author(s): 長田 重一
  • Organizer: 大塚製薬講演会
  • Invited
• [Presentation] Phosphatidylserine-Dependent Efferocytosis and Entosis (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: The 2019 Keystone Symposia on Why So Many Ways to Die? Apoptosis, Necroptosis, Pyroptosis and Beyond
  • Int'l Joint Research / Invited
• [Presentation] Flippases and scramblases for phosphatidylserine-exposure (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: The EMBO Workshop on "Cell Death, Immunity and Inflammation"
  • Int'l Joint Research / Invited
• [Presentation] Phosphatidylserine-dependent efferocytosis and entosis (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: The 27th Euroconference on Apoptosis "Cell Death and Regeneration"
  • Int'l Joint Research / Invited
• [Presentation] Mammalian Scramblases and Flippases at Plasma Membranes (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: Gordon Research Conference on Molecular and Cellular Biology of Lipids
  • Int'l Joint Research / Invited
• [Presentation] Phospholipid Flippase and Scramblase (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: IMS-JSI International Symposium on Immunology 2019
  • Int'l Joint Research / Invited
• [Presentation] Phospholipid Flippase and Scramblase (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: The 60th International Conference on the Bioscience of Lipids
  • Int'l Joint Research / Invited
• [Presentation] Phospholipid Flippase and Scramblase (2019)
  • Author(s): Shigekazu Nagata
  • Organizer: The 2nd Cell Death, Cell Stress and Metabolism Conference
  • Int'l Joint Research / Invited
• [Patent(Industrial Property Rights)] P2X7受容体発現調節剤 (2020)
  • Inventor(s): 長田重一、領田優太、瀬川勝盛
  • Industrial Property Rights Holder: 長田重一、領田優太、瀬川勝盛
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: PCT/JP2020/020308
  • Overseas
• [Patent(Industrial Property Rights)] P2X7受容体発現調節剤 (2019)
  • Inventor(s): 長田重一、領田優太、瀬川勝盛
  • Industrial Property Rights Holder: 長田重一、領田優太、瀬川勝盛
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 特願 2019-098049
• [Patent(Industrial Property Rights)] ATP11CまたはCDC50Aの阻害物質のスクリーニング方法 (2019)
  • Inventor(s): 長田重一、瀬川勝盛
  • Industrial Property Rights Holder: 長田重一、瀬川勝盛
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 特願 2019-082492（分割出願）
• [Patent(Industrial Property Rights)] ATP11CまたはCDC50Aの阻害物質のスクリーニング方法 (2015)
  • Inventor(s): 長田重一、瀬川勝盛
  • Industrial Property Rights Holder: 長田重一、瀬川勝盛
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 第6525212号
• [Patent(Industrial Property Rights)] Method of screening ATP11C or CDC50A inhibitor (2015)
  • Inventor(s): 長田重一、瀬川勝盛
  • Industrial Property Rights Holder: 長田重一、瀬川勝盛
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: US 10295528
  • Overseas