mechanistic analysis of cardiac functions by building G protein signal network

• Project/Area Number: 17079007
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyushu University
• Principal Investigator: KUROSE Hitoshi Kyushu University, 大学院・薬学研究院, 教授 (10183039)
• Co-Investigator(Kenkyū-buntansha): NISHIDA Motohiro 九州大学, 大学院・薬学研究院, 准教授 (90342641) ; NAKAYA Michio 九州大学, 大学院・薬学研究院, 助教 (80464387)
• Project Period (FY): 2005 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥74,200,000 (Direct Cost: ¥74,200,000); Fiscal Year 2009: ¥13,900,000 (Direct Cost: ¥13,900,000); Fiscal Year 2008: ¥13,900,000 (Direct Cost: ¥13,900,000); Fiscal Year 2007: ¥13,900,000 (Direct Cost: ¥13,900,000); Fiscal Year 2006: ¥24,500,000 (Direct Cost: ¥24,500,000); Fiscal Year 2005: ¥8,000,000 (Direct Cost: ¥8,000,000)
• Keywords: G蛋白質 / 心肥大 / 心不全 / 線維化 / 受容体 / 圧負荷 / ATP / 百日咳毒素 / アンジオテンシンII受容体 / TRPCチャネル / リン酸化 / ホスホジエステラーゼ5阻害剤 / cAMP依存性リン酸化酵素 / cGMP依存性リン酸化酵素 / プリン受容体 / パネキシン / 心機能 / 線維化促進因子 / G_q蛋白質 / アンジオテンシン受容体 / 伸展刺激 / Rho / G_12 / 13蛋白質 / アデノシン-三リン酸 / NFAT / ジアシルグリセロール / ホスホリパーゼC / 細胞内Ca^<2+>濃度 / 心肥大応答 / 新生仔心室筋細胞 / MAPキナーゼ / 活性酸素 / Gタンパク質 / 細胞内シグナリング / 低分子量G蛋白質

Research Abstract

The heart is hypertrophied when it is exposed to stresses such as hypertension. When the stress is not removed, the heart develops heart failure that does not properly provide the blood to peripheral tissues. Cardiac fibrosis is characterized by excessive deposition of extracellular matrix proteins, and is one of the causes of heart failure that contributes to the impairment of cardiac function, especially relaxation ability. So far, it is believed that fibrosis of various tissues including the heart is a secondary event following hypertrophy, and is regulated by the signaling pathway of angiotensin II (Ang II) and transforming growth factor-β (TGF-β). In order to examine the roles of heterotrimeric G12 family G protein (G_<α12>/G_<α13> : Gα12/13) in the heart, we used RGS domain of p115RhoGEF (p115-RGS) to selectively block G_<α12/13>. Transgenic mice (p115-Tg mice) expressing inhibitory p115-RGS only in cardiac myocytes shows pressure overload-induced fibrosis without affecting hyper trophy. As p115-RGS is selectively expressed in cardiac myocytes, p115RGS is selective inhibitor of heterotrimeric G_<α12/13> protein, and G protein is generally activated by G protein-coupled receptors (GPCRs), there should be a GPCR that triggers cardiac fibrosis when the heart is exposed to pressure overload. To identify a GPCR that regulates fibrosis, we used stretch treatment of neonatal cardiomyocytes, as it is thought that stretch treatment of cells in vitro mimics in vivo pressure overload. The activation of Rho downstream of G_<α12/13> by stretch treatment is initiated by ATP and UDP released from cardiac myocytes, as enzymes involved in ATP degradation or blockers of purine receptor inhibit stretch-induced Rho activation. Purine receptor-selective blockers reveal that the receptor is P2Y6. Furthermore, inhibition of G-protein-coupled P2Y_6 receptors in vivo inhibits pressure overload-induced cardiac fibrosis. These results suggest that activation of G_<α12/13> in cardiomyocytes by ATP and/or UDP-stimulated P2Y_6 receptor triggers fibrosis in pressure overload-induced cardiac fibrosis.
Pertussis toxin (PTX) is widely used as a specific tool that ADP-ribosylates Gi and Go (Gi/Go) and uncouples receptors from Gi/Go. To examine signaling pathways of angiotensin II receptor (AT1R) stimulation in cardiac fibroblasts, we found that PTX increases the expression of AT1R and enhances AT1R-mediated response. Microarray analysis shows that PTX increases the expression of interleukin (IL)-1β. Decreased expression of IL-1β by knockdown or antibody inhibits the PTX-treatment-stimulated enhancement of AT1R-mediated response. PTX increased the expression of IL-1β and AT1R through activation of a small GTP-binding protein Rac that stimulates NADPH oxidase-dependent production of reactive oxygen species. PTX-induced late but not an early phase of Rac activation is blocked by inhibition of IL-1β. It is known that PYX binds to Toll-like receptor 4 (TLR4). Knockdown of TLR4 inhibits PTX-induced Rac activation and enhancement of AT1R-mediated responses. However, inhibition of TLR4 does not affect PTX-mediated ADP-ribosylation of Gi/Go. Thus, PTX binds to at least two receptors: one is TLR4 that activates Rac and enhances AT1R responses, and another is the binding site that mediates entry of PTX into cells ADP-ribosylation of Gi/Go.

Research Products

• [Journal Article] Phosphorylation of TRPC6 channels at Thr69 is required for anti-hypertrophic effects of phosphodiesterase 5 inhibition (2010)
  • Author(s): Nishida, M., Watanabe, K., Sato, Y., Nakaya, M., Kitajima, N., Ide, T., Inoue, R., Kurose, H.
  • Journal Title: J Biol Chem. 286(17) Pages: 13244-13253
• [Journal Article] Pertussis toxin up-regulates angiotensin type 1 receptors through Toll-like receptor 4-mediated Rac activation. (2010)
  • Author(s): Nishida, M., Suda, R., Nagamatsu, Y., Tanabe, S., Onohara, N., Nakaya, M., Kanaho, Y., Shibata, T., Uchida, K., Sumimoto, H., Sato, Y., Kurose, H.
  • Journal Title: J Biol Chem. 285(20) Pages: 15268-15277
• [Journal Article] Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound. (2009)
  • Author(s): Kiyonaka, S., Kato, K., Nishida, M., Mio, K., Numaga, T., Sawaguchi, Y., Yoshida, T., Wakamori, M., Mori, E., Numata, T., Ishii, M., Takemoto, H., Ojida, A., Watanabe, K., Uemura, A., Kurose, H., Morii, T., Kobayashi. T., Sato, Y., Sato, C., Hamachi, I., Mori, Y.
  • Journal Title: Proc.Natl.Acad.Sci.USA 106 Pages: 5400-5405
  • Peer Reviewed
• [Journal Article] Amphotericin B-induced renal tubular cell injury is mediated by Na^+ Influx through ion-permeable pores and subsequent activation of mitogen-activated protein kinases and elevation of intracellular Ca^<2+> concentration. (2009)
  • Author(s): Yano T, Itoh Y, Kawamura E, Maeda A, Egashira N, Nishida M, Kurose H, Oishi R.
  • Journal Title: Antimicrob.Agents Chemother. 53 Pages: 1420-1426
  • Peer Reviewed
• [Journal Article] LPA1 receptors mediate stimulation, whereas LPA2 receptors mediate inhibition, of migration of pancreatic cancer cells in response to lysophosphatidic acid and malignant ascites. (2009)
  • Author(s): Komachi M, Tomura H, Malchinkhuu E, Tobo M, Mogi C, Yamada T, Kimura T, Kuwabara A, Ohta H, Im DS, Kurose H, Takeyoshi I, Sato K, Okajima F.
  • Journal Title: Carcinogenesis 30 Pages: 457-465
  • Peer Reviewed
• [Journal Article] P2Y6 receptor-Gα12/13 signaling in cardiomyocytes triggers pressure overload-induced cardiac fibrosis (2008)
  • Author(s): Nishida, M., Sato, Y., Uemura, A., Narita, Y., Tozaki-Saitoh, H., Nakaya, M., Ide, T., Suzuki, K., Inoue, K., Nagao, T., Kurose, H.
  • Journal Title: EMBO J. 27(23) Pages: 3104-3115
• [Journal Article] P2Y_6 receptor-Ga_(12/13) signaling in cardiomyocytes triggers pressure overload-induced cardiac fibrosis (2008)
  • Author(s): Nishida, M., Sato, Y., Uemura, A., Narita, Y., Tozaki-Saitoh, H., Nakay a, M., Ide, T., Suzuki, K., Inoue, K., Nagao, T., and Kurose, H.
  • Journal Title: EMBO Journal 27 Pages: 3104-3115
  • Peer Reviewed
• [Journal Article] Prokineticin Receptor-1 Induces Neovascularization and Epicardial-Deri ved Progenitor Cell Differentiation (2008)
  • Author(s): Urayama, K., Celia Guilini, C., Turkeri, G., Takir, S., Kurose, H., Messaddeq, N., Dierich, A., and Nebigil, C.G.
  • Journal Title: Arteriosclerosis, Thrombosis, and Vascular Biology 28 Pages: 841-849
  • Peer Reviewed
• [Journal Article] Induction of scavenger receptor class B type I is critical for simvastatin enhancement of high-density lipoprotein-induced anti-inflammatory actions in endothelial cells (2008)
  • Author(s): Kimura, T., Mogi, C., Tomura, H., Kuwabara, A., Im, D.S., Sato, K., K urose, H., Murakami, M., and Okajima, F.
  • Journal Title: Journal of Immunology 181 Pages: 7332-73340
  • Peer Reviewed
• [Journal Article] 機械的伸展刺激により活性化されるGタンパク質共役型受容体の解析 (2008)
  • Author(s): 西田基宏、上村綾、仲矢道雄、黒瀬等
  • Journal Title: 表面 46 Pages: 28-38
• [Journal Article] Gα12/13-mediated upregulation of TRPC6 negatively regulates endothelin-1-induced cardiac myofibroblast formation and collagen synthesis through NFAT activation (2007)
  • Author(s): Nishida, M., Onohara, N., Sato, Y., Suda, R., Ogushi, M., Tanabe, S., Inoue, R., Mori, Y., Kurose, H.
  • Journal Title: J. Biol. Chem. 282(32) Pages: 23117-23128
• [Journal Article] Gα_<12/13>-mediated upregulation of TRRC6 negatively regulates endothelin-1-induced cardiac myofibroblast formation and collagen synthesis through NFAT activation. (2007)
  • Author(s): Nishida, M.・Onohara, N.・Sato, Y.・Suda, R・Ogushi, M.・Tanabe, S.・Inoue, R.・Mori, Y.・Kurose, H.
  • Journal Title: Journal of Biological Chemiostry 282 Pages: 23117-23128
  • Peer Reviewed
• [Journal Article] The prokineticin receptor-1(GPR73) promotes cardiomyocyte survival and angiogenesis. (2007)
  • Author(s): Urayama, K.・Guilini, C.・Messaddeq, N.・Hu, K.・Steenman, M.・Kurose, H.・Ert, G.・Nebigil, C.G.
  • Journal Title: FASEB Journal 21 Pages: 2980-2993
  • Peer Reviewed
• [Journal Article] Heterotrimeric G protein Gα13-induced induction of cytokine mRNAs through two distinct pathways in cardiac fibroblasts. (2006)
  • Author(s): Nagamatsu, Y., Nishida, M., Onohara, N., Fukutomi, M., Maruyama, Y., Kobayashi, H., Sato, Y., Kurose, H.
  • Journal Title: J. Pharmacol. Sci. 101 Pages: 144-150
• [Journal Article] TRPC3 and TRPC6 are essential for angiotensin II-induced cardiac hypertrophy. (2006)
  • Author(s): Onohara, N., Nishida, N., Inoue, R., Kobayashi, H., Sumimoto, H., Sato, Y., Mori, Y., Nagao, T., Kurose, H.
  • Journal Title: EMBO J. 25(22) Pages: 5305-5316
• [Journal Article] TRPC3 and TRPC6 are essential for angiotensin II-induced cardiac hypertrophy. (2006)
  • Author(s): Onohara, N., Nishida, N., Inoue, R., Kobayashi, H., Sumimoto, H., Sato, Y., Mori, Y., Nagao, T., Kurose, H.
  • Journal Title: EMBO Journal 25(22) Pages: 5305-5316
• [Journal Article] Clathrin required for phosphorylation and internalization of β_2-adrenergic receptor by G protein-coupled receptor kinase 2 (GRK2). (2006)
  • Author(s): Mangmool, S., Haga, T., Kobayashi, H., Kim, K-M., Nakata, H., Nishida, M., Kurose, H.
  • Journal Title: Journal of Biological Chemistry 281(42) Pages: 31940-31949
• [Journal Article] Heterotrimeric G protein Gα_13-induced induction of cytokine mRNAs through two distinct oathwavs in cardiac fibroblasts. (2006)
  • Author(s): Naganatsu, Y., Nishida, M., Onohara, N., Fukutomi, M., Maruyama, Y., Kobayashi, H., Sato, Y., Kurose, H.
  • Journal Title: Journal of Pharmacological Sciences 101 Pages: 144-150
• [Journal Article] Sphingosine 1-phosphate receptors mediate stimulatory and inhibitory signalings for expression of adhesion molecules in endothelial cells. (2006)
  • Author(s): Kimura, T., Tomura, H., Mogi, C., Kuwabara, A., Ishiwara, M., Shibasawa, K., Sato, K., Ohwada, S., Im, D.S, Kurose, H., Ishizuka, T., Murakami, M., Okajima, F.
  • Journal Title: Cellular Signalling 18(6) Pages: 841-850
• [Journal Article] Thromboxane A_2 receptor-mediated G_<12/13>-dependent glial morphological change. (2006)
  • Author(s): Honma, S., Saika, M., Ohkubo, S., Kurose, H., Nakahata, N.
  • Journal Title: European Journal of Pharmacology 545(2-3) Pages: 100-108
• [Journal Article] Beta-blockers shows inverse agonism to a novel constitutively active mutant of β_1-adrenoceptor. (2006)
  • Author(s): Ahmed, M., Muntasir, H.A., Hossain, M., Isiguro, M., Komiyama, T., Muramatsu, I., Kurose, H., Nagatomo, T.
  • Journal Title: Journal of Pharmacological Sciences 102 Pages: 167-172
  • NAID: 10020345451
• [Journal Article] Gα12/13-mediated production of reactive oxygen species is critical for angiotensin receptor-induced NFAT activation in cardiac fibroblasts (2005)
  • Author(s): Fujii, T., Onohara, N., Maruyama, Y., Tanabe, S., Kobayashi, H., Fukutomi, M., Nagamatsu, Y., Nishihara, N., Inoue, R., Sumimoto, H., Shibasaki, F., Nagao, T., Nishida, M., Kurose, H.
  • Journal Title: J. Biol. Chem. 280(24) Pages: 23041-23047
• [Journal Article] Gα12/13- and reactive oxygen species-dependent activation of c-Jun NH2-terminal kinase and p38 MAPK by angiotensin receptor stimulation in rat neonatal cardiomyocytes. (2005)
  • Author(s): Nishida, M., Tanabe, S., Maruyama, Y., Mangmool S., Urayama K., Nagamatsu, Y., Takagahara, S., Turner, J.H., Kozasa, T., Kobayashi, H., Sato, Y., Kawanishi, T., Inoue, R., Nagao, T., Kurose, H.
  • Journal Title: J. Biol. Chem. 280(18) Pages: 18434-18441
• [Journal Article] Caveolae-independent activation of protein kinase A in rat neonatal myocytes. (2005)
  • Author(s): Fukutomi, M., Nishida, M., Maruyama, Y., Kobayashi, H., Kurose, H.
  • Journal Title: J. Pharmacol. Sci. 98(2) Pages: 168-174
  • NAID: 10025728483
• [Journal Article] Gα_<12/13>- and reactive species-dependent activation of c-Jun NH_2-terminal kinase and p38 MAPK by angiotensin receptor stimulation in rat neonatal cardiomyocytes. (2005)
  • Author(s): Nishida, M., Tanabe, S., Maruyama, Y., Mangmool S., Urayama K., Nagamatsu, Y., Takagahara, S., Turner J.H., Kozasa, T., Kobayashi, H., Sato, Y., Kawanishi, T., Inoue, R., Nagao, T., Kurose, H.
  • Journal Title: Journal of Biological Chemistry 280(18) Pages: 18434-18441
• [Journal Article] Gα_<12/13>-mediated production of reactive oxygen species is critical for angotensin receptor-induced NFAT activation in cardiac fibroblasts. (2005)
  • Author(s): Fujii, T., Onohara, N., Maruyama, Y., Tanabe, S., Kobayashi, H., Fukutomi, M., Nagamatsu, Y., Nishihara, N., Inoue, R., Sumimoto, H., Shibasaki, F., Nagao, T., Nishida, M., Kurose, H.
  • Journal Title: Journal of Biological Chemistry 280(24) Pages: 23041-23047
• [Journal Article] Caveolae-independent activation of protein kinase A in rat neonatal myocytes. (2005)
  • Author(s): Fukutomi, M., Nishida, M., Maruyama, Y., Kobayashi, H., Kurose, H.
  • Journal Title: Journal of Pharmacological Science 98(2) Pages: 168-174
  • NAID: 10025728483
• [Journal Article] G protein-coupled receptor signaling through Gq and JNK negatively regulates neural progenitor cell migration. (2005)
  • Author(s): Mizuno, N., Kokubu, H., Sato, M., Nishimura, A., Yamauchi, J., Kurose, H., Itoh, H.
  • Journal Title: Proceedings of National Academy of Sciences USA 102(35) Pages: 12365-12370
• [Journal Article] Thronboxane A_2 promotes interleukin-6 synthesis mediated by an activation of cyclic AMP-response element binding prptein in 1321N1 human astrocytome cells. (2005)
  • Author(s): Obara, Y., Kurose, H., Nakahata, N.
  • Journal Title: Molecular Pharmacology 68(3) Pages: 670-679
• [Journal Article] Down-regulation of Na^+/Ca^<2+> exchanger by fluvastatin in rat cardiomyoblast H9c2 cells : Involvement of RhoB in Na^+/Ca^<2+> exchanger mRNA stability. (2005)
  • Author(s): Maeda, S., Matsuoka, I., Iwamoto, T., Kurose, H., Kimura, J.
  • Journal Title: Molecular Pharmacology 68(2) Pages: 414-420
• [Presentation] 心臓の線維化 (2009)
  • Author(s): 黒瀬等
  • Organizer: 日本生化学会九州支部例会
  • Place of Presentation: 九州大学病院キャンパス コラボステーション
  • Year and Date: 2009-05-16
• [Presentation] 線維化とプリン受容体 (2009)
  • Author(s): 黒瀬等
  • Organizer: 日本薬学会第129年会
  • Place of Presentation: 京都国際会議場・京都
  • Year and Date: 2009-03-26
• [Presentation] 線維化とプリン受容体 (2009)
  • Author(s): 黒瀬等
  • Organizer: 日本薬学会第129年会
  • Place of Presentation: 京都国際会議場
  • Year and Date: 2009-03-26
• [Presentation] アンジオテンシン受容体の調節機構 (2008)
  • Author(s): 黒瀬等、西田基宏、仲矢道雄、須田玲子、大串真理子
  • Organizer: 第81回日本薬理学会年会
  • Place of Presentation: 横浜パシフィコ・横浜
  • Year and Date: 2008-03-17
• [Presentation] βアドレナリン受容体遮断薬の薬理 (2007)
  • Author(s): 黒瀬等
  • Organizer: 第2回臨床薬理学会/薬理学会共催シンポジウム
  • Place of Presentation: 栃木県総合文化センター・宇都宮
  • Year and Date: 2007-11-28
• [Presentation] 心臓の線維化のトリガーとなるG_<12/13>蛋白質共役型受容体の同定 (2007)
  • Author(s): 西田基弘、黒瀬 等
  • Organizer: 第6回生命科学研究会
  • Place of Presentation: 京成ホテルミラマーレ、千葉
  • Year and Date: 2007-07-06
• [Presentation] 受容体発現を制御するGタンパク質 (2007)
  • Author(s): 黒瀬等
  • Organizer: 薬学会第127年会
  • Place of Presentation: 富山国際会議場など・富山
  • Year and Date: 2007-03-29
• [Presentation] Racを介したアンジオテンシン受容体の発現調節 (2006)
  • Author(s): 黒瀬等
  • Organizer: 第34回薬物活性シンポジウム・第115回日本薬理学会関東部会
  • Place of Presentation: 高崎シティギャラリー、高崎市庁舎会議室・群馬
  • Year and Date: 2006-09-29
• [Presentation] リガンドに依存したシグナリングの差異 (2006)
  • Author(s): 黒瀬等
  • Organizer: 第126回薬学会年会
  • Place of Presentation: 仙台国際センター・仙台
  • Year and Date: 2006-03-29
• [Remarks]
  • URL: http://chudoku.phar.kyushu-u.ac.jp/
• [Remarks]
  • URL: http://210.233.60.66/~chudoku/