Novel Mechanisms of Oxidative Stress Response in Heart Failure

心力衰竭氧化应激反应的新机制

基本信息

批准号：
10930191
负责人：
QIN M CHEN
金额：
$ 62.53万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-23 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10930191
关键词：
5&apos Untranslated Regions Angioplasty Animal Model Antioxidants Binding Binding Proteins Bioinformatics Biological Assay Biological Markers Cardiac Cardiac Myocytes Categories Cell Death Cell Proliferation Cells Chemicals Clinical Complex Coronary Artery Bypass Coronary artery Cytoprotection Data Data Science Data Set Disease Drug Metabolic Detoxication Event Excision Genes Heart failure Human Infarction Inflammation Knock-out Knowledge Label Laboratory Finding Laboratory Research Life Literature Mediating Messenger RNA Meta-Analysis Metabolism Methods Methylation Methyltransferase Mitochondria Molecular Conformation Myocardial Myocardial Infarction Myocardial Ischemia Myocardium NF-E2-related factor 2 National Heart, Lung, and Blood Institute Operative Surgical Procedures Oxidants Oxidative Stress Patients Polymerase Postoperative Period Protein Inhibition Proteins Proteomics Publishing RNA methylation RNA-Binding Proteins Reaction Reading Regulatory Element Reperfusion Therapy Resources Ribonucleases Ribosomes Role Site Statistical Computing Stress Stress-Induced Protein System Techniques Testing Time Trans-Omics for Precision Medicine Translating Translation Initiation Translations Validation biological adaptation to stress cohort cost effective therapy human disease improved ischemic cardiomyopathy mortality myocardial injury nanopore novel programs protein protein interaction recruit repaired tissue degeneration tissue repair transcription factor transcriptome sequencing transcriptomics

项目摘要

Novel Mechanisms of Oxidative Stress Response in Heart Failure A large volume of literature indicates an association of oxidative stress with end stage heart failure. Myocardial ischemia or infarction is the most common cause of heart failure, which is difficult to treat and costly to manage, and has a high mortality rate. A deficiency in oxidant removal or damage repair systems in the myocardium may explain the progression of heart failure. At the cellular level, low to mild levels of oxidative stress activate Nrf2 transcription factor to turn on the expression of antioxidant and detoxification genes. We have found that de novo Nrf2 protein translation serves as an important mechanism for Nrf2 activation under oxidative stress. How certain proteins, such as Nrf2, can be selectively translated when cardiac cells encounter oxidative stress remains largely unknown. When investigating the ribosomes using quantitative LC-MS/MS based proteomics, we have recently discovered that YTHD2, a YTH domain containing N6 methyladenosine (m6A) binding protein, increased its association with the ribosomes during oxidative stress. We have found that YTHD2 increases the binding to Nrf2 mRNA and have detected an increase of m6A in Nrf2 mRNA with oxidative stress. Knocking out YTHD2 blocked oxidants from inducing Nrf2 protein. These data lead us to hypothesize that YTHD2 reading of m6A in the Nrf2 5’ untranslated region (5’UTR) serves as a passport for de novo Nrf2 protein translation under oxidative stress. The loss of Nrf2 protein translation mechanisms may contribute to end stage heart failure. Aim 1 will test that oxidative stress causes Nrf2 mRNA methylation at specific sites and m6A landscape change due to activation of METTL3 methylase. Aim 2 will test whether YTHD2 binding causes a Nrf2 5’UTR conformation change and recruitment of a translation initiation complex for de novo Nrf2 protein translation. Aim 3 will demonstrate the losses of stress induced protein translation machinery, regulated mRNA methylation, and Nrf2 mediated cytoprotection in end stage heart failure patients. Our study will advance the knowledge pertaining to a novel discovery involving RNA methylation for de novo protein translation in oxidative stress response. This project will utilize transcriptomic data from heart failure patients to validate mechanistic findings from laboratory research for implications in real life human disease.

心力衰竭中氧化应激反应的新机制大量文献表明氧化应激与最终阶段心力衰竭的关联。心肌缺血或梗塞是心力衰竭的最常见原因，这很难治疗，并且管理高昂的管理，并具有高死亡率。心肌中氧化剂清除或损伤修复系统的缺乏可能解释心力衰竭的进展。在细胞水平上，低至轻度的氧化应激激活NRF2 转录因子打开抗氧化剂和解毒基因的表达。我们发现DE Novo NRF2蛋白翻译是氧化应激下NRF2激活的重要机制。如何当心脏细胞遇到氧化应激时，可以选择性地翻译某些蛋白质，例如NRF2 仍然是未知的。当使用基于定量LC-MS/MS的蛋白质组学研究核糖体时，我们最近发现，YTHD2是一个含有N6甲基腺苷（M6A）结合蛋白的YTH结构域，在氧化应激期间增加了与核糖体的关联。我们发现ythd2增加了与NRF2 mRNA的结合，并检测到具有氧化应激的NRF2 mRNA中M6a的增加。淘汰 YTHD2从诱导的NRF2蛋白中阻断了氧化。这些数据使我们假设ythd2读取 NRF2 5'非翻译区（5'UTR）中的M6A用作NRF2 NRF2蛋白转换的护照在氧化应激下。 NRF2蛋白翻译机制的丧失可能导致终点心脏衰竭。 AIM 1将测试氧化应激会在特定位点和M6A上引起NRF2 mRNA甲基化由于Mettl3甲基化酶的激活而引起的景观变化。 AIM 2将测试YTHD2结合是否引起A NRF2 5'UTR构象的变化和募集了从头NRF2蛋白的翻译启动复合物翻译。 AIM 3将证明应力诱导的蛋白质翻译机制的损失，调节mRNA 甲基化和NRF2介导的末期心力衰竭患者的细胞保护。我们的研究将推进与新发现有关的知识，该发现涉及氧化中从头蛋白转化的RNA甲基化压力反应。该项目将利用心力衰竭患者的转录组数据来验证机理实验室研究的结果对现实生活中的人类疾病的影响。