ETS2-dependent control in cardiomyocyte ischemia/reperfusion injury

ETS2 依赖性控制心肌细胞缺血/再灌注损伤

• 批准号: 10501545
• 负责人: THOMAS G GILLETTE
• 金额: $ 61.79万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10501545
• 关键词: Acute; Apoptotic; Binding; Calcineurin; Cardiac; Cardiac Myocytes; Cardiac development; Cell Death; Connexin 43; Coronary; Coronary Arteriosclerosis; Data; EFRAC; ETS2 gene; Extracellular Signal Regulated Kinases; Family; Future; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Growth; Heart; Heart Hypertrophy; Heart failure; Hypertrophy; Immunoprecipitation; In Vitro; Injury; Ischemia; Ischemic Preconditioning; Ligation; Link; MAP2K1 gene; MAPK3 gene; Mass Spectrum Analysis; Mediating; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Modeling; Molecular; Morbidity - disease rate; Myocardial Infarction; Myocardial Ischemia; Myocardium; Pathogenesis; Pathologic; Pathway interactions; Physiological; Play; Predisposition; Proteins; Reperfusion Injury; Reperfusion Therapy; Reporting; Risk Factors; Role; Signal Pathway; Signal Transduction; Stress; Testing; Up-Regulation; Ventricular Remodeling; cardioprotection; cell growth; cell injury; gain of function; gap junction channel; genome-wide; in vivo; ischemic injury; member; mortality; novel; nuclear factors of activated T-cells; pressure; promoter; recruit; response; transcription factor; transcriptome sequencing

Project Summary/Abstract Heart failure is a leading cause of morbidity and mortality around the world, and the leading cause of heart failure with reduced ejection fraction (HFrEF) is coronary artery disease. The mitogen-activated protein kinases (MAPK) are an essential signal transduction cascade that play a central role in both cell growth and cell death. The MEK1-ERK1/2 branch of the MAPK pathway has been shown to promote both physiologic and pathologic growth in the heart, as well as protect against apoptotic cell death after ischemia/reperfusion (I/R) injury. We have previously demonstrated that the transcription factor ETS2, a member of E26 transformation-specific sequence (ETS)-domain family, is phosphorylated and activated by Erk1/2 upon hypertrophic stimulation. Going forward, our preliminary data reveal that cardiomyocyte- specific loss of ETS2 results in increased susceptibility to ischemic injury in both I/R and permanent ligation (myocardial infarction) models of heart failure. Connexin43 (Cx43), the predominant gap junction channel- forming protein in cardiomyocytes, has been suggested to play a role in both ischemic damage and ischemic preconditioning. Our preliminary data show that ETS2 activates Cx43 transcription and that Cx43 is downregulated in the absence of ETS2. We will test the hypothesis that the ERK1/2/ETS2 pathway protects against I/R injury in part through the upregulation of Cx43. Aim 1: To determine the role of the ERK1/2-ETS2 pathway in I/R injury. Our preliminary data suggest a model in which ETS2 protects against I/R injury. We will confirm and extend this using loss- and gain-of- function approaches in vivo and in vitro. We will track the timing of ETS2 activation by ERK and the response of each acutely and in long-term remodeling. Aim 2: To determine the impact of ETS2 on Cx43 expression and function in I/R injury. Our data suggest that Cx43 plays a protective role in ischemic injury. Our preliminary data also suggest that ETS2 is a direct transcriptional regulator of Cx43 gene expression. We will confirm and extend these findings using both loss- and gain-of-function approaches. We will also determine the role of ETS2 in Cx43-mediated cardioprotection in IPC. Aim 3: To determine the downstream targets and interactors of ETS2 under conditions of cardiac stress. We will profile genome-wide cardiac gene expression using RNAseq and ChIPseq to determine ETS2 downstream gene targets in both acute and long-term I/R injury. We will use immunoprecipitation and mass spectrometry to unveil novel protein interactions.

项目摘要/摘要 心力衰竭是世界各地发病率和死亡率的主要原因，是 射血分数（HFREF）的心力衰竭是冠状动脉疾病。有丝分裂原激活 蛋白激酶（MAPK）是一个必不可少的信号转导级联，在两个细胞中都起着核心作用 生长和细胞死亡。已显示MAPK途径的MEK1-ERK1/2分支都可以促进 心脏的生理和病理生长，并防止凋亡细胞死亡 缺血/再灌注（I/R）损伤。我们以前已经证明了转录因子ETS2，A E26转化特异性序列（ETS）域家族的成员被磷酸化并被激活 肥厚刺激后的ERK1/2。展望未来，我们的初步数据表明心肌细胞 - ETS2的具体损失导致I/R和永久连接的缺血性损伤的敏感性增加 （心肌梗死）心力衰竭模型。 Connexin43（CX43），主要间隙连接通道 - 在心肌细胞中形成蛋白质，已被建议在缺血性损害和缺血性中发挥作用 预处理。我们的初步数据表明，ETS2激活CX43转录，CX43是 在没有ETS2的情况下下调。我们将测试ERK1/2/ETS2途径保护的假设 通过上调CX43，反对I/R损伤。 目标1：确定ERK1/2-ETS2途径在I/R损伤中的作用。我们的初步数据暗示 ETS2防止I/R损伤的模型。我们将使用损失和收益来确认并扩展这一点 功能在体内和体外接近。我们将通过ERK跟踪ETS2激活的时机和响应 每个急性和长期重塑。 目标2：确定ETS2对I/R损伤中CX43表达和功能的影响。我们的数据 表明CX43在缺血性损伤中起保护作用。我们的初步数据还表明ETS2是 CX43基因表达的直接转录调节剂。我们将使用 损失和功能收益的方法。我们还将确定ETS2在CX43介导的 IPC中的心脏保护。 目标3：在心脏条件下确定ETS2的下游目标和相互作用者 压力。我们将使用RNASEQ和CHIPSEQ介绍全基因组的心脏基因表达 急性和长期I/R损伤中的ETS2下游基因靶标。我们将使用免疫沉淀和 质谱法揭示了新型蛋白质相互作用。