Grb2-associated binder 1 is essential for cardioprotection against ischemia/reperfusion injury

Abstract We have shown recently that endothelial Grb-2-associated binder 1 (Gab1), an intracellular scaffolding adaptor, has a protective effect against limb ischemia via mediating angiogenic signaling pathways. However, the role of Gab1 in cardiac ischemia/reperfusion (I/R) injury remains unknown. In this study, we show that Gab1 is required for cardioprotection against I/R injury. I/R injury led to remarkable phosphorylation of Gab1 in cardiomyocytes. Compared with controls, the mice with cardiomyocyte-specific deletion of Gab1 gene (CGKO mice) exhibited an increase in infarct size and a decrease in cardiac function after I/R injury. Consistently, in hearts of CGKO mice subjected to I/R, the activation of caspase 3 and myocardial apoptosis was markedly enhanced whereas the activation of protein kinase B (Akt) and mitogen-activated protein kinase (MAPK), which are critical for cardiomyocyte survival, was attenuated. Oxidative stress is regarded as a major contributor to myocardial I/R injury. To examine the role of Gab1 in oxidative stress directly, isolated adult cardiomyocytes were subject to oxidant hydrogen peroxide and the cardioprotective effects of Gab1 were confirmed. Furthermore, we found that the phosphorylation of Gab1 and Gab1-mediated activation of Akt and MAPK by oxidative stress was suppressed by ErbB receptor and Src kinase inhibitors, accompanied by an increase in apoptotic cell death. In conclusion, our results suggest that Gab1 is essential for cardioprotection against I/R oxidative injury via mediating survival signaling.

[1]  N. Dhalla,et al.  Status of myocardial antioxidants in ischemia-reperfusion injury. , 2000, Cardiovascular research.

[2]  T. Yue,et al.  Inhibition of extracellular signal-regulated kinase enhances Ischemia/Reoxygenation-induced apoptosis in cultured cardiac myocytes and exaggerates reperfusion injury in isolated perfused heart. , 2000, Circulation research.

[3]  R. Kitsis,et al.  Akt promotes survival of cardiomyocytes in vitro and protects against ischemia-reperfusion injury in mouse heart. , 2000, Circulation.

[4]  G. Heusch,et al.  Mitogen-activated protein kinases in the heart , 2001, Naunyn-Schmiedeberg's Archives of Pharmacology.

[5]  Y. Zou,et al.  Oxidative stress activates extracellular signal-regulated kinases through Src and Ras in cultured cardiac myocytes of neonatal rats. , 1997, The Journal of clinical investigation.

[6]  L. Leng,et al.  Cardiac macrophage migration inhibitory factor inhibits JNK pathway activation and injury during ischemia/reperfusion. , 2009, The Journal of clinical investigation.

[7]  Jie Wu,et al.  Role of Gab1 in UV-Induced c-Jun NH2-Terminal Kinase Activation and Cell Apoptosis , 2004, Molecular and Cellular Biology.

[8]  Ji Young Kim,et al.  Endothelial Grb2-Associated Binder 1 Is Crucial for Postnatal Angiogenesis , 2011, Arteriosclerosis, thrombosis, and vascular biology.

[9]  G. Heusch,et al.  Ischemic Postconditioning in Pigs: No Causal Role for Risk Activation , 2008, Circulation research.

[10]  D. Yellon,et al.  Reperfusion injury salvage kinase and survivor activating factor enhancement prosurvival signaling pathways in ischemic postconditioning: two sides of the same coin. , 2011, Antioxidants & redox signaling.

[11]  J. Massagué.  TGF-beta signal transduction. , 1998, Annual review of biochemistry.

[12]  P. Chan,et al.  Differential phosphorylation of the docking protein Gab1 by c-Src and the hepatocyte growth factor receptor regulates different aspects of cell functions , 2010, Oncogene.

[13]  Bernd Giese,et al.  Targeting phosphoinositide 3-kinase: moving towards therapy. , 2008, Biochimica et biophysica acta.

[14]  P. Doevendans,et al.  MEK1-ERK2 Signaling Pathway Protects Myocardium From Ischemic Injury In Vivo , 2004, Circulation.

[15]  W. Birchmeier,et al.  Essential Role of Gab1 for Signaling by the C-Met Receptor in Vivo , 2000, The Journal of cell biology.

[16]  Marina Holgado-Madruga,et al.  Gab1 Is an Integrator of Cell Death versus Cell Survival Signals in Oxidative Stress , 2003, Molecular and Cellular Biology.

[17]  R. Jennings,et al.  Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. , 1986, Circulation.

[18]  Yibin Wang.  Mitogen-activated protein kinases in heart development and diseases. , 2007, Circulation.

[19]  G. Heusch,et al.  Inconsistent relation of MAPK activation to infarct size reduction by ischemic preconditioning in pigs. , 2000, American journal of physiology. Heart and circulatory physiology.

[20]  D. Yellon,et al.  Myocardial reperfusion injury. , 2007, The New England journal of medicine.

[21]  M. Shibuya,et al.  Grb-2–associated binder 1 (Gab1) regulates postnatal ischemic and VEGF-induced angiogenesis through the protein kinase A–endothelial NOS pathway , 2011, Proceedings of the National Academy of Sciences.

[22]  Yan Liu,et al.  The gift of Gab , 2002, FEBS letters.

[23]  R. Hajjar,et al.  Akt Activation Preserves Cardiac Function and Prevents Injury After Transient Cardiac Ischemia In Vivo , 2001, Circulation.

[24]  E. Topol.  Current Status and Future Prospects for Acute Myocardial Infarction Therapy , 2003, Circulation.

[25]  J. Penninger,et al.  The molecular scaffold Gab2 is a crucial component of RANK signaling and osteoclastogenesis , 2005, Nature Medicine.

[26]  J. C. Pratt,et al.  Essential role for Gab2 in the allergic response , 2001, Nature.

[27]  G. Heusch.  Cardioprotection: chances and challenges of its translation to the clinic , 2013, The Lancet.

[28]  T. Hirano,et al.  Requirement of Gab2 for mast cell development and KitL/c-Kit signaling. , 2002, Blood.

[29]  A. Godwin,et al.  A Grb2-associated docking protein in EGF- and insulin-receptor signalling , 1996, Nature.

[30]  P. Kang,et al.  Mechanisms and inhibitors of apoptosis in cardiovascular diseases. , 2011, Current pharmaceutical design.

[31]  A. Hevener,et al.  Deletion of Gab1 in the liver leads to enhanced glucose tolerance and improved hepatic insulin action , 2005, Nature Medicine.

[32]  A. Mayr,et al.  PI3Kγ Protects from Myocardial Ischemia and Reperfusion Injury through a Kinase-Independent Pathway , 2010, PloS one.

[33]  William Wijns,et al.  ACUTE MYOCARDIAL INFARCTION: REPERFUSION TREATMENT , 2002, Heart.

[34]  塩山 渉.  Docking protein is an essential component of postnatal angiogenesis after ischemia via HGF/c-Met signaling , 2011 .

[35]  T. Suter,et al.  Neuregulin-1 beta attenuates doxorubicin-induced alterations of excitation-contraction coupling and reduces oxidative stress in adult rat cardiomyocytes. , 2006, Journal of molecular and cellular cardiology.

[36]  A. Lochner,et al.  Kinases and phosphatases in ischaemic preconditioning: a re-evaluation , 2010, Basic Research in Cardiology.

[37]  A. Takeshita,et al.  Direct evidence for increased hydroxyl radicals originating from superoxide in the failing myocardium. , 2000, Circulation research.

[38]  N. Holbrook,et al.  Epidermal Growth Factor Receptor-dependent Akt Activation by Oxidative Stress Enhances Cell Survival* , 2000, The Journal of Biological Chemistry.

[39]  B. Kobilka,et al.  Dual modulation of cell survival and cell death by beta(2)-adrenergic signaling in adult mouse cardiac myocytes. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[40]  N. LeBrasseur,et al.  Cardiac Endothelial Cells Regulate Reactive Oxygen Species-induced Cardiomyocyte Apoptosis through Neuregulin-1β/erbB4 Signaling* , 2004, Journal of Biological Chemistry.

[41]  Yi Zhu,et al.  Metformin attenuates cardiac fibrosis by inhibiting the TGFbeta1-Smad3 signalling pathway. , 2010, Cardiovascular research.

[42]  R. Bolli,et al.  Myocardial Protection at a Crossroads: The Need for Translation Into Clinical Therapy , 2004, Circulation research.

[43]  S. Sollott,et al.  Role of Glycogen Synthase Kinase-3β in Cardioprotection , 2009, Circulation research.

[44]  M. Febbraio,et al.  Deletion of macrophage migration inhibitory factor protects the heart from severe ischemia-reperfusion injury: a predominant role of anti-inflammation. , 2011, Journal of molecular and cellular cardiology.

[45]  G. Heusch,et al.  Cardioprotection: nitric oxide, protein kinases, and mitochondria. , 2008, Circulation.

[46]  M. Jacobson.  Reactive oxygen species and programmed cell death. , 1996, Trends in biochemical sciences.

[47]  D. Yellon,et al.  Conditioning the whole heart--not just the cardiomyocyte. , 2012, Journal of molecular and cellular cardiology.

[48]  Xi-zheng Zhang,et al.  Osteoblastic molecular scaffold Gab1 is required for maintaining bone homeostasis , 2010, Journal of Cell Science.

[49]  A. Rosenzweig,et al.  Convergent signal transduction pathways controlling cardiomyocyte survival and function: the role of PI 3-kinase and Akt. , 2005, Journal of molecular and cellular cardiology.

[50]  T. Hirano,et al.  The role of Gab family scaffolding adapter proteins in the signal transduction of cytokine and growth factor receptors , 2003, Cancer science.

[51]  B. Neel,et al.  The "Gab" in signal transduction. , 2003, Trends in cell biology.

[52]  R. Gottlieb,et al.  The Interplay between Pro-Death and Pro-Survival Signaling Pathways in Myocardial Ischemia/Reperfusion Injury: Apoptosis Meets Autophagy , 2006, Cardiovascular Drugs and Therapy.

[53]  W. Lederer,et al.  Calcium sparks: elementary events underlying excitation-contraction coupling in heart muscle. , 1993, Science.

[54]  N. Xu,et al.  Targeted Disruption of Smad4 in Cardiomyocytes Results in Cardiac Hypertrophy and Heart Failure , 2005, Circulation research.

[55]  T. Kodama,et al.  Gab family proteins are essential for postnatal maintenance of cardiac function via neuregulin-1/ErbB signaling. , 2007, The Journal of clinical investigation.

[56]  R. Hajjar,et al.  Adenoviral gene transfer of activated phosphatidylinositol 3'-kinase and Akt inhibits apoptosis of hypoxic cardiomyocytes in vitro. , 1999, Circulation.

[57]  T. Hirano,et al.  Role of Gab1 in Heart, Placenta, and Skin Development and Growth Factor- and Cytokine-Induced Extracellular Signal-Regulated Kinase Mitogen-Activated Protein Kinase Activation , 2000, Molecular and Cellular Biology.

[58]  G. Heusch,et al.  p38 MAP kinase is a mediator of ischemic preconditioning in pigs. , 2002, Cardiovascular research.

[59]  E. Murphy,et al.  Mechanisms underlying acute protection from cardiac ischemia-reperfusion injury. , 2008, Physiological reviews.

[60]  H. Jo,et al.  Redox-sensitive Akt and Src regulate coronary collateral growth in metabolic syndrome. , 2009, American journal of physiology. Heart and circulatory physiology.