Secretome analysis for the search for novel danger signal alarmins in ATP-stimulated macrophages

• Project/Area Number: 25450521
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied molecular and cellular biology
• Research Institution: National Institute of Agrobiological Sciences
• Principal Investigator: TAKENOUCHI Takato 国立研究開発法人農業生物資源研究所, 動物生体防御研究ユニット, 上級研究員 (20292518)
• Co-Investigator(Kenkyū-buntansha): TSUKIMOTO Mitsutoshi 東京理科大学, 薬学部薬学科, 講師 (70434040)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: アラーミン / マクロファージ / P2X7受容体 / GAPDH / 非典型的分泌機構 / エキソソーム

Outline of Final Research Achievements

Alarmins are endogeneous molecules that act as danger signals to promote inflammation or innate immune response. Since they are mainly secreted from macrophage-related cells, we performed P2X7 receptor-secretome analysis using ATP-stimulated mouse microglial cells, known as brain macrophages, to search for novel alarmins. We have identified 640 different proteins as candidate molecules that may act as alarmins. Among them, we demonstrated the unconventional secretion pathway and innate immunomodulatory functions of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in microglial cells. The findings suggest that GAPDH secreted from macrophages may act as a mediator of innate immune system.

Research Products

• [Journal Article] ATP-triggered unconventional secretion of GAPDH from macrophages: Its putative role as a modulator of inflammation. (2015)
  • Author(s): Takenouchi T., Tsukimoto M., Hashimoto M., Kitani H.
  • Journal Title: Macrophage Volume: 2
  • DOI: 10.14800/macrophage.1083
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Extracellular ATP induces unconventional release of glyceraldehyde-3-phosphate dehydrogenase from microglial cells (2015)
  • Author(s): Takenouchi T, Tsukimoto M, Iwamaru Y, Sugama S, Sekiyama K, Sato M, Kojima S, Hashimoto M, Kitani H
  • Journal Title: Immunol Lett Volume: 167 Issue: 2 Pages: 116-124
  • DOI: 10.1016/j.imlet.2015.08.002
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Extracellular ATP induces P2X7 receptor activation in mouse Kupffer cells, leading to release of IL-1β, HMGB1, and PGE2, decreased MHC class I expression and necrotic cell death. (2015)
  • Author(s): Toki Y, Takenouchi T, Harada H, Tanuma S, Kitani H, Kojima S, Tsukimoto M.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 458(4) Issue: 4 Pages: 771-776
  • DOI: 10.1016/j.bbrc.2015.02.011
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Inflammasome activation by danger signals: extracellular ATP and pH (2014)
  • Author(s): Takenouchi T, Tsukimoto M, Hashimoto M, Kitani H
  • Journal Title: Inflammasome Volume: 1 Issue: 1 Pages: 76-80
  • DOI: 10.2478/infl-2014-0008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Extracellular ATP does not induce P2X7 receptor-dependent responses in cultured renal- and liver-derived swine macrophages (2014)
  • Author(s): Takenouchi T, Suzuki S, Shinkai H, Tsukimoto M, Sato M, Uenishi H, Kitani H
  • Journal Title: Results in Immunology Volume: 4 Pages: 62-67
  • DOI: 10.1016/j.rinim.2014.07.002
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Purinergic signaling via P2X7 receptor mediates IL-1β production in Kupffer cells exposed to silica nanoparticle (2014)
  • Author(s): Kojima S, Negishi Y, Tsukimoto M, Takenouchi T, Kitani H, Takeda K
  • Journal Title: Toxicology Volume: 321 Pages: 13-20
  • DOI: 10.1016/j.tox.2014.03.008
  • Peer Reviewed
• [Journal Article] Purinergic signaling via P2Y receptors up-mediates IL-6 production by liver macrophages/Kupffer cells (2014)
  • Author(s): Ishimaru M, Yusuke N, Tsukimoto M, Harada H, Takenouchi T, Kitani H, Kojima S
  • Journal Title: The Journal of Toxicological Sciences Volume: 39 Pages: 413-423
  • NAID: 130004447125
  • Peer Reviewed
• [Presentation] ミクログリアにおけるGAPDHの非典型的放出機構と自然免疫応答調節 (2016)
  • Author(s): 竹之内敬人, 月本光俊, 佐藤充, 木谷裕
  • Organizer: 日本農芸化学会2016年度大会
  • Place of Presentation: 札幌
  • Year and Date: 2016-03-30
• [Presentation] ブタ腎臓および肝臓マクロファージの単離・培養とP2X7受容体機能解析 (2015)
  • Author(s): 竹之内敬人, 鈴木俊一, 新開浩樹, 月本光俊, 佐藤充, 上西博英, 木谷裕
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山
  • Year and Date: 2015-03-28
• [Book] EMERGING DRUGS AND TARGETS FOR PARKINSON´S DISEASE (2014)
  • Author(s): Takenouchi T, Sekiyama K, Sugama S, Iwamaru Y, Kitani H, Hashimoto M
  • Total Pages: 402
  • Publisher: RSC publishing