Live cell imaging of regulated necrosis
Project/Area Number |
16K01378
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Biomedical engineering/Biomaterial science and engineering
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Research Institution | Toho University |
Principal Investigator |
MURAI Shin 東邦大学, 医学部, 助教 (90287540)
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Co-Investigator(Kenkyū-buntansha) |
中野 裕康 東邦大学, 医学部, 教授 (70276476)
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Research Collaborator |
YAMAGUCHI Yoshifumi
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Project Period (FY) |
2016-04-01 – 2019-03-31
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Project Status |
Completed (Fiscal Year 2018)
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Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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Keywords | MLKL / RIPK3 / Necroptosis / FRET / ネクロプトーシス / HMGB1 / LCI-S / 1分子FRET / 計画的ネクローシス / 1分子FRET / ライブセルイメージング |
Outline of Final Research Achievements |
Regulated necrosis is a regulated form of necrosis that depends on RIPK3 and MLKL. To monitor the induction of regulated necrosis, we have designed FRET biosensor (KLN) which inserted the linker sequence derived from MLKL between CFP and YFP. Since KLN expression suppressed the cell proliferation, the KLN linker sequence was modified. As the result, FRET biosensor without cytotoxicity was developed and termed as SMART. SMART can monitor regulated necrosis, but not apoptosis or necrosis in vitro, indicating that SMART is a specific sensor to regulated necrosis. FRET was not observed by the knockdown of RIPK3 expression or the inhibition of RIPK3 activity, suggesting that SMART can monitor RIPK3 activity which is required the induction of regulated necrosis. Since the unphosphorylated SMART mutant can also monitor RIPK3 activation by its FRET, it was concluded that FRET of SMART is required for the interaction of RIPK3 but not phosphorylation by RIPK3.
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Academic Significance and Societal Importance of the Research Achievements |
疾患により損傷をうけた組織では、アポトーシスや計画的ネクローシスに加え、貪食されずに残った死細胞による二次的ネクローシス、ウイルス感染によるパイロトーシスや膜脂質の酸化によるフェロトーシスなど様々な細胞死が混在していることが予想される。病態の増悪を抑制するためには、それぞれの細胞死に特異的な阻害剤が有用である。本研究の成果により、計画的ネクローシスを他の細胞死と区別して検出することで、効果的な治療薬の選択が可能となる。さらに薬剤投与による計画的ネクローシスの抑制効果の解析が可能であるため、ドラッグ・スクリーニングによる新規治療薬の開発をはじめとする医療面への貢献も期待できる。
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Report
(4 results)
Research Products
(42 results)
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[Journal Article] A FRET biosensor for necroptosis uncovers two different modes of the release of DAMPs.2018
Author(s)
Murai S, Yamaguchi Y, Shirasaki Y, Yamagishi M, Shindo R, Hildebrand JM, Miura R, Nakabayashi O, Totsuka M, Tomida T, Adachi-Akahane S, Uemura S, Silke J, Yagita H, Miura M, Nakano H
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Journal Title
Nat Commun
Volume: 9
Issue: 1
Pages: 4457-4457
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Depletion of myeloid cells exacerbates hepatitis and induces an aberrant increase in histone H3 in mouse serum2017
Author(s)
Piao, X. Yamazaki, S. Komazawa-Sakon, S. Miyake, S. Nakabayashi, O. Kurosawa, T. Mikami, T. Tanaka, M. Van Rooijen, N. Ohmuraya, M. Oikawa, A. Kojima, Y. Kakuta, S. Uchiyama, Y. Tanaka, M. Nakano, H.
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Journal Title
Hepatology
Volume: 65
Issue: 1
Pages: 237-252
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] HTLV-1 Tax induces formation of the active macromolecular IKK complex by generating Lys63- and Met1-linked hybrid polyubiquitin chains.2017
Author(s)
Shibata Y, Tokunaga F, Goto E, Komatsu G, Gohda J, Saeki Y, Tanaka K, Takahashi H, Sawasaki T, Inoue S, Oshiumi H, Seya T, Nakano H, Tanaka Y, Iwai K, and Inoue J.
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Journal Title
PLoS Pathog.
Volume: 13
Issue: 12
Pages: e1006162-e1006162
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] Novel method to rescue a lethal phenotype through integration of target gene onto the X-chromosome2016
Author(s)
Sakata, K. Araki, K. Nakano, H. Nishina, T. Komazawa-Sakon, S. Murai, S. Lee, G. E. Hashimoto, D. Suzuki, C. Uchiyama, Y. Notohara, K. Gukovskaya, A. S. Gukovsky, I. Yamamura, K. I. Baba, H. Ohmuraya, M.
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Journal Title
Sci Rep
Volume: 6
Issue: 1
Pages: 37200-37200
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Presentation] FRETバイオセンサーによるネクロプトーシス実行の1細胞イメージング2018
Author(s)
村井 晋, 山口 良文, 白崎 義隆, 進藤 綾大, 中林 修, Hildebrand Joanne, 冨田 太一郎, 赤羽 悟美, Silke John, 三浦 正幸, 中野 裕康
Organizer
第91回日本生化学会大会
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[Presentation] 細胞死制御因子cFLIPの肝細胞特異的欠損マウスを用いた肝障害モデルにおけるクッパー細胞および骨髄由来細胞の役割2016
Author(s)
朴 雪花, 山﨑 創, 駒澤-左近 幸子, 三宅 早苗, 中林 修, 田中 稔, 大村谷 昌樹, 及川 彰, 角田 宗一郎, 内山 安男, 田中 正人, 中野 裕康
Organizer
第89回日本生化学会大会
Place of Presentation
東北大学(宮城県・仙台)
Year and Date
2016-09-26
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