Role of miR-574 driver and passenger strands in cardiac hypertrophy

miR-574驱动链和过客链在心脏肥大中的作用

• 批准号: 9336417
• 负责人: Peng Yao
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-19 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336417
• 关键词: Address; Adrenergic Agonists; Adult; Biology; C57BL/6 Mouse; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cell Death; Cell physiology; Cells; Chronic; Coculture Techniques; Complex; DNA; Development; Drug Targeting; Echocardiography; Elements; Etiology; Exhibits; Fibroblasts; Fibrosis; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Genetic Translation; Goals; Heart; Heart Hypertrophy; Heart failure; Heterogeneous-Nuclear Ribonucleoprotein L; Human; Hypertrophy; Hypoxia; In Vitro; Injection of therapeutic agent; Ischemia; Isoproterenol; Knockout Mice; Knowledge; Lead; Link; Mediating; Messenger RNA; MicroRNAs; Molecular; Morbidity - disease rate; Mus; Neonatal; Pathway interactions; Phenotype; Phosphorylation; Play; Prevention approach; Preventive Intervention; Process; Regulation; Research; Ribonucleases; Role; Seeds; Serum; Starvation; Stress; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Therapeutic Intervention; Tissues; Transcription Factor 3; Transfection; Translations; United States; Untranslated RNA; Untranslated Regions; Vascular Endothelial Growth Factors; Wild Type Mouse; activating transcription factor; base; constriction; exosome; improved; in vivo; insight; intercellular communication; locked nucleic acid; mimetics; mortality; mouse model; myocyte-specific enhancer-binding factor 2; novel; prevent; small molecule; subcutaneous; targeted treatment; therapeutic miRNA; therapeutic target; transcription factor; transcriptome

Project Summary: Cardiovascular disease (CVD) is the leading cause of morbidity and mortality worldwide. Better understanding of the pathological mechanisms underlying CVD will improve preventive and therapeutic interventions. One major etiology of CVD is dysregulation of cardiac gene expression by transcription factors (TFs) and microRNAs (miRNAs). miRNAs are small non-coding RNAs involved in gene expression regulation. TFs are usually poor small-molecule drug targets. However, miRNAs can act as unique therapeutic agents because they can target multiple TFs in the same pathological pathway. Most precursor miRNAs (pre-miRNAs) are processed to generate an active driver strand miRNA; the complementary passenger strand is usually degraded. Intriguingly, pre-miR-574 produces two functional strands—miR-574-5p and miR-574-3p. We found that both strands of miR-574 play a synergistic role to protect hearts from cardiac hypertrophy. We showed that miR-574-5p was induced in chronic human heart failure tissues versus normal hearts. We found that miR-574-5p is up-regulated under hypertrophic and ischemic stress and silences the expression of three pro-hypertrophy TFs myocyte enhancer factor–2 factors (Mef2a/2c/2d) and prevents cardiomyocyte (CM) hypertrophy. Transfection of miR-574-5p mimics in mouse cardiomyocytes (CM) protects the cells from isoproterenol (ISO, a β-adrenergic agonist)-induced hypertrophy and hypoxia-serum starvation-induced cell death. miR-574 knockout mice exhibit an advanced cardiac hypertrophy phenotype associated with increased fibrosis and enlarged CM, compared to wild-type mice after chronic ISO injection. We also showed that the passenger strand miR-574-3p is co-upregulated with miR-574-5p but captured by hypoxia-activated phospho-hnRNP L (heterogeneous nuclear ribonucleoprotein L). miR-574-3p is secreted from CM via exosomes and targets fibroblasts to reduce fibrosis. Our research goal is to address fundamental questions in miRNA biology using miR-574-5p/3p as archetypal molecules in the cardiac system (differential regulation, mechanistic diversity, and functional relationship between driver and passenger strands), and to treat HF by developing miRNA-based therapeutics that target TFs. Our central hypothesis is: In CM under hypertrophic and ischemic stress, differential regulation of the driver strand miR-574-5p and passenger strand miR-574-3p integrate cardiac gene expression and modulate pathological cardiac hypertrophy and remodelling. We will test this hypothesis by pursuing two aims. Aim 1 will test the hypothesis that miR-574-5p regulates the expression of Mef2 TFs and other cardiac genes via miRISC, thereby modulates cardiac hypertrophy and remodelling. Aim 2 will test the hypothesis that in CM ischemia promotes miR-574 transcription via specific TFs and activates capture of miR-574-3p by P-hnRNP L followed by exosome secretion and intercellular communication.

项目摘要： 心血管疾病（CVD）是全球发病率和死亡率的主要原因。更好的 了解CVD背后的病理机制将改善预防性和 治疗干预措施。 CVD的主要病因是心脏基因表达的失调 通过转录因子（TFS）和microRNA（miRNA）。 miRNA是小的非编码RNA 参与基因表达调节。 TF通常是小分子药物靶标。 但是，miRNA可以充当独特的治疗剂，因为它们可以瞄准多个TF 相同的病理途径。大多数前体miRNA（前MIRNA）都被处理以生成 活跃的驱动器链mirna；互补的乘客链通常会退化。 有趣的是，前MIR-574产生两个功能链-MIR-574-5P和miR-574-3p。我们发现 miR-574的两条链都起着协同作用，可以保护心脏免受心脏肥大的影响。我们 结果表明，在慢性人类心力衰竭组织与正常心脏中诱导miR-574-5p。 我们发现miR-574-5p在肥厚和缺血性压力下被上调，沉默 表达三个亲毛TFS肌细胞增强因子– 2因子（MEF2A/2C/2D）和 防止心肌细胞（CM）肥大。 miR-574-5p Mimics在小鼠中转染 心肌细胞（CM）保护细胞免受异丙肾上腺素（ISO，β-肾上腺素激动剂）诱导的 肥大和缺氧 - 阴性饥饿诱导的细胞死亡。 mir-574淘汰小鼠暴露了 与纤维化增加和商业增加有关的晚期心脏肥大表型， 与慢性ISO注射后的野生型小鼠相比。我们还表明了乘客股 miR-574-3p与miR-574-5p共同调节，但被低氧激活的磷酸化磷酸化捕获 L（异质核核糖核蛋白L）。 miR-574-3p通过外泌体分泌CM， 靶向成纤维细胞以减少纤维化。我们的研究目标是解决基本问题 miRNA生物学使用miR-574-5p/3p作为心脏系统中的原型分子（差异） 驾驶员与乘客之间的监管，机械多样性和功能关系 链），并通过开发靶向TFS的基于miRNA的疗法来治疗HF。我们的中心 假设是：在肥厚和缺血性压力下CM中，驱动器的差异调节 Strand miR-574-5p和乘客链mir-574-3p综合心脏基因表达和 调节病理心脏肥大和重塑。我们将通过 追求两个目标。 AIM 1将测试miR-574-5p调节MEF2表达的假设 TFS和其他心脏基因通过Mirisc，从而调节心脏肥大和重塑。 AIM 2将检验以下假设，即在CM缺血中促进MiR-574通过特定TFS促进miR-574转录 并激活P-HNRNP L对miR-574-3p的捕获，然后进行外泌体分泌和 细胞间沟通。