Elucidation of molecualr mechanism underlying cardiac hibernation by reactive persulfide species

2016 Fiscal Year Annual Research Report

• Project/Area Number: 15K14959
• Research Institution: Okazaki Research Facilities, National Institutes of Natural Sciences
• Principal Investigator: 西田 基宏 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡崎統合バイオサイエンスセンター, 教授 (90342641)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 活性イオウ / 心臓 / 冬眠化 / 活性酸素 / 心不全

Outline of Annual Research Achievements

我々はこれまで、生体内に存在する求核性の高い「活性イオウ種」が、活性酸素や環境汚染物質などの親電子性の高い化合物（親電子物質）を適切に解毒・排泄する重要なシステムとして働くことを明らかにしてきた。本研究では、①活性イオウ種の分子実体の解明、②活性イオウ種の生成・代謝制御機構、③心筋の生理機能との関連、に着目した検討を行った。まず、ポリ硫黄鎖を検出できる蛍光指示薬（SSP）やTag-switch-Tag法を用いて細胞内ポリ硫黄の測定を行ったところ、レドックス感受性の高いシグナルタンパク質に含まれるシステインポリ硫黄鎖が活性イオウ種の分子実体であること、および親電子物質によるポリ硫黄鎖の枯渇がタンパク質の不可逆的な酸化的翻訳後修飾や多量体化を誘発することで、マウス・ラット心筋細胞のミトコンドリア過剰分裂や細胞老化を誘導しうることを見出した。また、タンパク質にシステインパースルフィドを供給する酵素がシステインtRNA合成酵素（CARS2）であることも新たに見出した。CARS2をノックダウンあるいは過剰発現させたラット新生児心筋細胞を用いてCa2+ハンドリングを調べたところ、脱分極誘発性のCa2+スパークの形や大きさに異常は見当たらなかった。ところが、CARS2欠損細胞においてミトコンドリア分裂促進Gタンパク質dynamin-related protein 1 (Drp1)の活性が有意に上昇しており、この細胞にシステインパースルフィド生成活性のみ保持したCARS2変異体（tRNA合成酵素活性欠損）を発現させることで、増加していたDrp1活性が完全に低下することを確認した。以上の結果から、細胞内の活性イオウ種は、心筋において、ミトコンドリア分裂タンパク質などの安定性や酵素活性を負に調節する役割を担うことが明らかとなった。

Research Products

• [Int'l Joint Research] Sabah大学医学部(マレーシア)
  • Country Name: MALAYSIA
  • Counterpart Institution: Sabah大学医学部
• [Int'l Joint Research] Mahidol大学(タイ)
  • Country Name: THAILAND
  • Counterpart Institution: Mahidol大学
• [Int'l Joint Research] Catholic University of Argentina(アルゼンチン)
  • Country Name: ARGENTINA
  • Counterpart Institution: Catholic University of Argentina
• [Int'l Joint Research] NIEHS(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: NIEHS
• [Journal Article] Purinergic P2Y6 receptors: a new therapeutic target of age-dependent hypertension (2017)
  • Author(s): Sunggip C, Nishimura A, Shimoda K, Numaga-Tomita T, Tsuda M, and Nishida M. M
  • Journal Title: Pharmacological Research Volume: 120 Pages: -
  • DOI: http://dx.doi.org/10.1016/j.phrs.2017.03.013
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MiR30-GALNT1/2 Axis-Mediated Glycosylation Contributes to the Increased Secretion of Inactive Human Prohormone for Brain Natriuretic Peptide (proBNP) From Failing Hearts. (2017)
  • Author(s): Nakagawa Y, Nishikimi T, Kuwahara K, Fujishima A, Oka S, Tsutamoto T, Kinoshita H, Nakao K, Cho K, Inazumi H, Okamoto H, Nishida M, Kato T, Fukushima H, Yamashita JK, Wijnen WJ, Creemers EE, Kangawa K, Minamino N, Nakao K, Kimura T. M.
  • Journal Title: J Am Heart Assoc. Volume: Feb 10; 6(2). Pages: -
  • DOI: doi: 10.1161/JAHA.116.003601.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Redox regulation of electrophilic signaling by reactive persulfides in cardiac cells. (2017)
  • Author(s): Nishida M, Nishimura A, Matsunaga T, Motohashi H, Kasamatsu S, Akaike T
  • Journal Title: Free Radic Biol Med. Volume: Jan 18. Pages: -
  • DOI: doi: 10.1016/j.freeradbiomed.2017.01.024.
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] TRPC3 participates in angiotensin II type 1 receptor-dependent stress-induced slow increase in intracellular Ca2+ concentration in mouse cardiomyocytes. (2017)
  • Author(s): Yamaguchi Y, Iribe G, Kaneko T, Takahashi K, Numaga-Tomita T, Nishida M, Birnbaumer L, Naruse K.
  • Journal Title: J Physiol Sci. Volume: Jan 19. Pages: -
  • DOI: doi: 10.1007/s12576-016-0519-3.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TRPC3-GEF-H1 axis mediates pressure overload-induced cardiac fibrosis. (2016)
  • Author(s): Numaga-Tomita T, Kitajima N, Kuroda T, Nishimura A, Miyano K, Yasuda S, Kuwahara K, Sato Y, Ide T, Birnbaumer L, Sumimoto H, Mori Y, Nishida M.
  • Journal Title: Sci Rep. Volume: Dec 19; 6:39383. Pages: -
  • DOI: doi: 10.1038/srep39383.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TRPC3 positively regulates reactive oxygen species driving maladaptive cardiac remodeling. (2016)
  • Author(s): Kitajima N, Numaga-Tomita T, Watanabe M, Kuroda T, Nishimura A, Miyano K, Yasuda S, Kuwahara K, Sato Y, Ide T, Birnbaumer L, Sumimoto H, Mori Y, Nishida M
  • Journal Title: Sci Rep. Volume: Nov 11; 6:37001. Pages: -
  • DOI: doi: 10.1038/srep37001.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Methylmercury, an environmental electrophile capable of activation and disruption of the Akt/CREB/Bcl-2 signal transduction pathway in SH-SY5Y cells. (2016)
  • Author(s): Unoki T, Abiko Y, Toyama T, Uehara T, Tsuboi K, Nishida M, Kaji T, Kumagai Y.
  • Journal Title: Sci Rep. Volume: Jun 30; 6:28944. Pages: -
  • DOI: doi: 10.1038/srep28944.
  • Peer Reviewed / Open Access
• [Journal Article] Synthesis of radioiodinated probes to evaluate the biodistribution of a potent TRPC3 inhibitor. (2016)
  • Author(s): Hagimori M, Murakami T, Shimizu K, Nishida M, Ohshima T, Mukai T.
  • Journal Title: Med. Chem. Comm. Volume: 7(5) Pages: 1003-1006
  • DOI: DOI: 10.1039/c6md00023a.
  • Peer Reviewed / Open Access
• [Journal Article] Redox signaling regulated by an electrophilic cyclic nucleotide and reactive cysteine persulfides. (2016)
  • Author(s): Fujii S, Sawa T, Nishida M, Ihara H, Ida T, Motohashi H, Akaike T.
  • Journal Title: Arch Biochem Biophys. Volume: 595 Pages: 140-146
  • DOI: doi: 10.1016/j.abb.2015.11.008.
  • Peer Reviewed / Open Access
• [Journal Article] Redox signaling regulated by electrophiles and reactive sulfur species. (2016)
  • Author(s): Nishida M, Kumagai Y, Ihara H, Fujii S, Motohashi H, Akaike T
  • Journal Title: J Clin Biochem Nutr. Volume: 58(2) Pages: 91-98
  • DOI: doi: 10.3164/jcbn.15-111.
  • Peer Reviewed / Open Access
• [Presentation] Myocardial early senescence mediated by mitochondria-cytoskeleton interaction (2016)
  • Author(s): Nishida M
  • Organizer: The 39th Annual Meeting of the Molecular Biology Society of Japan (international symposium)
  • Place of Presentation: Pacifico Yokohama, Yokohama, Kanagawa
  • Year and Date: 2016-12-01 – 2016-12-02
  • Int'l Joint Research
• [Presentation] Regulation of cardiac oxygen remodeling via electrophilic modification of Drp1 (2016)
  • Author(s): Nishida M
  • Organizer: The 89th Annual Meeting of the Japanese Biochemical Society (international symposium)
  • Place of Presentation: Sendai International Conference Center, Sendai, Miyagi
  • Year and Date: 2016-09-25 – 2016-09-27
  • Int'l Joint Research
• [Presentation] 活性イオウによる心筋のストレス適応・不適応の分子制御 (2016)
  • Author(s): 西田基宏
  • Organizer: 日本獣医学会比較薬理毒性分科会
  • Place of Presentation: 日本大学、神奈川県藤沢市
  • Year and Date: 2016-09-06 – 2016-09-06
  • Invited
• [Presentation] TRPC channels in cardiovascular stress resilience (2016)
  • Author(s): Nishida M
  • Organizer: International and Interdisciplinary Symposium 2016 “Towards a New Era of Cardiovascular Research”
  • Place of Presentation: Tokyo Medical and Dental University, Bunkyo-ku, Tokyo
  • Year and Date: 2016-07-11 – 2016-07-13
  • Int'l Joint Research / Invited
• [Presentation] Redox regulation of G proteins in cardiac remodeling (2016)
  • Author(s): Nishida M
  • Organizer: The 9th International Conference on the Biology, Chemistry, and Therapeutic Applications of Nitric Oxide
  • Place of Presentation: Sendai International Conference Center, Sendai, Miyagi
  • Year and Date: 2016-05-20 – 2016-05-22
  • Int'l Joint Research
• [Remarks] 心循環シグナル研究部門
  • URL: http://www.nips.ac.jp/circulation/