RhoA protects the mouse heart against ischemia/reperfusion injury.
Authors:
Journal: The Journal of clinical investigation
Publication Type: Journal Article
Date: 2011
DOI: PMC3148727
ID: 21747165
Abstract
The small GTPase RhoA serves as a nodal point for signaling through hormones and mechanical stretch. However, the role of RhoA signaling in cardiac pathophysiology is poorly understood. To address this issue, we generated mice with cardiomyocyte-specific conditional expression of low levels of activated RhoA (CA-RhoA mice) and demonstrated that they exhibited no overt cardiomyopathy. When challenged by in vivo or ex vivo ischemia/reperfusion (I/R), however, the CA-RhoA mice exhibited strikingly increased tolerance to injury, which was manifest as reduced myocardial lactate dehydrogenase (LDH) release and infarct size and improved contractile function. PKD was robustly activated in CA-RhoA hearts. The cardioprotection afforded by RhoA was reversed by PKD inhibition. The hypothesis that activated RhoA and PKD serve protective physiological functions during I/R was supported by several lines of evidence. In WT mice, both RhoA and PKD were rapidly activated during I/R, and blocking PKD augmented I/R injury. In addition, cardiac-specific RhoA-knockout mice showed reduced PKD activation after I/R and strikingly decreased tolerance to I/R injury, as shown by increased infarct size and LDH release. Collectively, our findings provide strong support for the concept that RhoA signaling in adult cardiomyocytes promotes survival. They also reveal unexpected roles for PKD as a downstream mediator of RhoA and in cardioprotection against I/R.
Chemical List
- L-Lactate Dehydrogenase|||protein kinase D|||Protein Kinase C|||RhoA protein, mouse|||rho GTP-Binding Proteins|||rhoA GTP-Binding Protein
Reference List
- Sander EE, Collard JG. Rho-like GTPases: their role in epithelial cell-cell adhesion and invasion. Eur J Cancer. 1999;35(14):1905–1911. doi: 10.1016/S0959-8049(99)00293-2.|||Ai S, et al. Rho-Rho kinase is involved in smooth muscle cell migration through myosin light chain phosphorylation-dependent and independent pathways. Atherosclerosis. 2001;155(2):321–327. doi: 10.1016/S0021-9150(00)00585-2.|||Zhang S, et al. RhoA regulates G1-S progression of gastric cancer cells by modulation of multiple INK4 family tumor suppressors. Mol Cancer Res. 2009;7(4):570–580. doi: 10.1158/1541-7786.MCR-08-0248.|||Ueyama T, et al. Activated RhoA stimulates c-fos gene expression in myocardial cells. . Circ Res. 1997;81(5):672–678.|||Miyamoto S, Del Re DP, Xiang SY, Zhao X, Florholmen G, Brown JH. Revisited and revised: is RhoA always a villain in cardiac pathophysiology? J Cardiovasc Transl Res. 2010;3(4):330–343. doi: 10.1007/s12265-010-9192-8.|||Del Re DP, Miyamoto S, Brown JH. RhoA /Rho kinase upregulate Bax to activate a mitochondrial death pathway and induce cardiomyocyte apoptosis. J Biol Chem. 2007;282(11):8069–8078. doi: 10.1074/jbc.M604298200.|||Sah VP, et al. Cardiac-specific overexpression of RhoA results in sinus and atrioventricular nodal dysfunction and contractile failure. J Clin Invest. 1999;103(12):1627–1634. doi: 10.1172/JCI6842.|||Kontaridis MI, et al. Deletion of Ptpn11 (Shp2) in cardiomyocytes causes dilated cardiomyopathy via effects on the extracellular signal-regulated kinase/mitogen-activated protein kinase and RhoA signaling pathways. Circulation. 2008;117(11):1423–1435. doi: 10.1161/CIRCULATIONAHA.107.728865.|||Phrommintikul A, et al. Effects of a Rho kinase inhibitor on pressure overload induced cardiac hypertrophy and associated diastolic dysfunction. Am J Physiol Heart Circ Physiol. 2008;294(4):H1804–H1814. doi: 10.1152/ajpheart.01078.2007.|||Higashi M, et al. Long-term inhibition of Rho-kinase suppresses angiotensin II-induced cardiovascular hypertrophy in rats in vivo: effect on endothelial NAD(P)H oxidase system. Circ Res. 2003;93(8):767–775. doi: 10.1161/01.RES.0000096650.91688.28.|||Shi J, Zhang L, Wei L. Rho-kinase in development and heart failure: insights from genetic models. Pediatr Cardiol. 2011;32(3):297–304. doi: 10.1007/s00246-011-9920-0.|||Dong M, Yan BP, Liao JK, Lam YY, Yip GW, Yu CM. Rho-kinase inhibition: a novel therapeutic target for the treatment of cardiovascular diseases. Drug Discov Today. 2010;15(15–16):622–629. doi: 10.1016/j.drudis.2010.06.011.|||Del Re DP, Miyamoto S, Brown JH. Focal adhesion kinase as a RhoA-activable signaling scaffold mediating Akt activation and cardiomyocyte protection. J Biol Chem. 2008;283(51):35622–35629. doi: 10.1074/jbc.M804036200.|||Krijnen PA, et al. Inhibition of Rho-ROCK signaling induces apoptotic and non-apoptotic PS exposure in cardiomyocytes via inhibition of flippase. J Mol Cell Cardiol. 2010;49(5):781–790. doi: 10.1016/j.yjmcc.2010.07.017.|||Mozzicato S, Joshi BV, Jacobson KA, Liang BT. Role of direct RhoA-phospholipase D1 interaction in mediating adenosine-induced protection from cardiac ischemia. FASEB J. 2004;18(2):406–408.|||Sanbe A, Gulick J, Hanks MC, Liang Q, Osinska H, Robbins J. Reengineering inducible cardiac-specific transgenesis with an attenuated myosin heavy chain promoter. Circ Res. 2003;92(6):609–616. doi: 10.1161/01.RES.0000065442.64694.9F.|||Sugimoto N, Takuwa N, Yoshioka K, Takuwa Y. Rho-dependent, Rho kinase-independent inhibitory regulation of Rac and cell migration by LPA1 receptor in Gi-inactivated CHO cells. Exp Cell Res. 2006;312(10):1899–1908. doi: 10.1016/j.yexcr.2006.02.020.|||Murga C, Zohar M, Teramoto H, Gutkind JS. Rac1 and RhoG promote cell survival by the activation of PI3K and Akt, independently of their ability to stimulate JNK and NF-kappaB. Oncogene. 2002;21(2):207–216. doi: 10.1038/sj.onc.1205036.|||Zhang B, Zhang Y, Shacter E. Rac1 inhibits apoptosis in human lymphoma cells by stimulating Bad phosphorylation on Ser-75. Mol Cell Biol. 2004;24(14):6205–6214. doi: 10.1128/MCB.24.14.6205-6214.2004.|||Palaniyandi SS, Sun L, Ferreira JC, Mochly-Rosen D. Protein kinase C in heart failure: a therapeutic target? Cardiovasc Res. 2009;82(2):229–239. doi: 10.1093/cvr/cvp001.|||Rey O, Reeve JR, Jr, Zhukova E, Sinnett-Smith J, Rozengurt E. G protein-coupled receptor-mediated phosphorylation of the activation loop of protein kinase D: dependence on plasma membrane translocation and protein kinase Cepsilon. J Biol Chem. 2004;279(33):34361–34372. doi: 10.1074/jbc.M403265200.|||Tan M, Xu X, Ohba M, Ogawa W, Cui MZ. Thrombin rapidly induces protein kinase D phosphorylation, and protein kinase C delta mediates the activation. J Biol Chem. 2003;278(5):2824–2828. doi: 10.1074/jbc.M211523200.|||Brandlin I, Eiseler T, Salowsky R, Johannes FJ. Protein kinase C(mu) regulation of the JNK pathway is triggered via phosphoinositide-dependent kinase 1 and protein kinase C(epsilon). J Biol Chem. 2002;277(47):45451–45457. doi: 10.1074/jbc.M205299200.|||Matthews SA, Rozengurt E, Cantrell D. Characterization of serine 916 as an in vivo autophosphorylation site for protein kinase D/Protein kinase Cmu. J Biol Chem. 1999;274(37):26543–26549. doi: 10.1074/jbc.274.37.26543.|||Aktories K, Rosener S, Blaschke U, Chhatwal GS. Botulinum ADP-ribosyltransferase C3: purification of the enzyme and characterization of the ADP-ribosylation reaction in platelet membranes. Eur J Biochem. 1988;172(2):445–450. doi: 10.1111/j.1432-1033.1988.tb13908.x.|||Sharlow ER, et al. Potent and selective disruption of protein kinase D functionality by a benzoxoloazepinolone. J Biol Chem. 2008;283(48):33516–33526. doi: 10.1074/jbc.M805358200.|||Brown JH, Del Re DP, Sussman MA. The Rac and Rho hall of fame: a decade of hypertrophic signaling hits. Circ Res. 2006;98(6):730–742. doi: 10.1161/01.RES.0000216039.75913.9e.|||Sah VP, Hoshijima M, Chien KR, Brown JH. Rho is required for Galphaq and alpha1-adrenergic receptor signaling in cardiomyocytes. Dissociation of Ras and Rho pathways. J Biol Chem. 1996;271(49):31185–31190. doi: 10.1074/jbc.271.49.31185.|||Morissette MR, Sah VP, Glembotski CC, Brown JH. The Rho effector, PKN, regulates ANF gene transcription in cardiomyocytes through a serum response element. Am J Physiol Heart Circ Physiol. 2000;278(6):H1769–H1774.|||Zhang YM, et al. Targeted deletion of ROCK1 protects the heart against pressure overload by inhibiting reactive fibrosis. FASEB J. 2006;20(7):916–925. doi: 10.1096/fj.05-5129com.|||Yuan J, Slice LW, Rozengurt E. Activation of protein kinase D by signaling through Rho and the alpha subunit of the heterotrimeric G protein G13. J Biol Chem. 2001;276(42):38619–38627. doi: 10.1074/jbc.M105530200.|||Cowell CF, Yan IK, Eiseler T, Leightner AC, Doppler H, Storz P. Loss of cell-cell contacts induces NF-kappaB via RhoA-mediated activation of protein kinase D1. J Cell Biochem. 2009;106(4):714–728. doi: 10.1002/jcb.22067.|||Liu GS, Cohen MV, Mochly-Rosen D, Downey JM. Protein kinase C-epsilon is responsible for the protection of preconditioning in rabbit cardiomyocytes. J Mol Cell Cardiol. 1999;31(10):1937–1948. doi: 10.1006/jmcc.1999.1026.|||Baines CP, et al. Protein kinase Cepsilon interacts with and inhibits the permeability transition pore in cardiac mitochondria. Circ Res. 2003;92(8):873–880. doi: 10.1161/01.RES.0000069215.36389.8D.|||Hamid SA, Bower HS, Baxter GF. Rho kinase activation plays a major role as a mediator of irreversible injury in reperfused myocardium. Am J Physiol Heart Circ Physiol. 2007;292(6):H2598–H2606. doi: 10.1152/ajpheart.01393.2006.|||Haudek SB, et al. Rho kinase-1 mediates cardiac fibrosis by regulating fibroblast precursor cell differentiation. Cardiovasc Res. 2009;83(3):511–518. doi: 10.1093/cvr/cvp135.|||Guilluy C, et al. The Rho exchange factor Arhgef1 mediates the effects of angiotensin II on vascular tone and blood pressure. Nat Med. 2010;16(2):183–190. doi: 10.1038/nm.2079.|||Masumoto A, Mohri M, Shimokawa H, Urakami L, Usui M, Takeshita A. Suppression of coronary artery spasm by the Rho-kinase inhibitor fasudil in patients with vasospastic angina. Circulation. 2002;105(13):1545–1547. doi: 10.1161/hc1002.105938.|||Zhou Q, Gensch C, Liao JK. Rho-associated coiled-coil-forming kinases (ROCKs): potential targets for the treatment of atherosclerosis and vascular disease. Trends Pharmacol Sci. 2011;32(3):167–173. doi: 10.1016/j.tips.2010.12.006.|||Nohria A, et al. Rho kinase inhibition improves endothelial function in human subjects with coronary artery disease. Circ Res. 2006;99(12):1426–1432. doi: 10.1161/01.RES.0000251668.39526.c7.|||Noma K, et al. ROCK1 mediates leukocyte recruitment and neointima formation following vascular injury. J Clin Invest. 2008;118(5):1632–1644. doi: 10.1172/JCI29226.|||Sagi SA, Seasholtz TM, Kobiashvilla M, Wilson BA, Toksoz D, Brown JH. Physical and functional interactions of Gαq with Rho and its exchange factors. . J Biol Chem. 2001;276(18):15445–15452. doi: 10.1074/jbc.M008961200.|||Means CK, et al. Sphingosine 1-phosphate S1P2 and S1P3 receptor-mediated Akt activation protects against in vivo myocardial ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol. 2007;292(6):H2944–H2951. doi: 10.1152/ajpheart.01331.2006.|||Parsons SA, et al. Genetic loss of calcineurin blocks mechanical overload-induced skeletal muscle fiber type switching but not hypertrophy. J Biol Chem. 2004;279(25):26192–26200. doi: 10.1074/jbc.M313800200.