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: 10674020
• 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/10674020
• 关键词: Acute; Apoptotic; Binding; Calcineurin; Cardiac; Cardiac Myocytes; Cell Death; ChIP-seq; Connexin 43; Coronary; Coronary Arteriosclerosis; Data; Development; 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 Reperfusion Injury; Myocardium; Pathogenesis; Pathologic; Pathway interactions; Phosphorylation; Phosphotransferases; 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; loss of function; member; mortality; novel; nuclear factors of activated T-cells; pressure; promoter; protective pathway; 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.

项目概要/摘要 心力衰竭是全世界发病率和死亡率的主要原因，也是 射血分数降低的心力衰竭（HFrEF）是冠状动脉疾病。有丝分裂原激活 蛋白激酶 (MAPK) 是一种重要的信号转导级联，在细胞和细胞中发挥着核心作用 生长和细胞死亡。 MAPK 通路的 MEK1-ERK1/2 分支已被证明可以促进两者 心脏的生理和病理生长，以及防止细胞凋亡后死亡 缺血/再灌注（I/R）损伤。我们之前已经证明转录因子 ETS2 E26 转化特异性序列 (ETS) 结构域家族的成员，被磷酸化并被激活 肥大刺激后的 Erk1/2。展望未来，我们的初步数据显示心肌细胞- ETS2 的特异性缺失导致 I/R 和永久结扎中对缺血性损伤的易感性增加 （心肌梗塞）心力衰竭模型。 Connexin43 (Cx43)，主要的间隙连接通道 在心肌细胞中形成蛋白质，已被认为在缺血性损伤和缺血性损伤中发挥作用 预处理。我们的初步数据表明 ETS2 激活 Cx43 转录并且 Cx43 在没有 ETS2 的情况下下调。我们将检验 ERK1/2/ETS2 通路保护的假设 部分通过 Cx43 的上调来对抗 I/R 损伤。 目标 1：确定 ERK1/2-ETS2 通路在 I/R 损伤中的作用。我们的初步数据表明 ETS2 可以防止 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 体内和体外功能方法。我们将跟踪 ERK 激活 ETS2 的时间和响应 每一个都在急剧和长期的重塑中。 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在缺血性损伤中发挥保护作用。我们的初步数据还表明 ETS2 是 Cx43 基因表达的直接转录调节因子。我们将使用以下方法确认并扩展这些发现 功能丧失和功能获得方法。我们还将确定 ETS2 在 Cx43 介导中的作用 IPC 中的心脏保护。 目标 3：确定心脏条件下 ETS2 的下游靶标和相互作用因子 压力。我们将使用 RNAseq 和 ChIPseq 分析全基因组心脏基因表达，以确定 ETS2 下游基因靶向急性和长期 I/R 损伤。我们将使用免疫沉淀法和 质谱法揭示新的蛋白质相互作用。