8OH-G miR-483 contributes to the aging-accelerated atherosclerosis

8OH-G miR-483 有助于衰老加速的动脉粥样硬化

• 批准号: 10452425
• 负责人: Ming He
• 金额: $ 23.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452425
• 关键词: 3' Untranslated Regions; Adenine; Adrenergic Agonists; Affect; Aging; Antiinflammatory Effect; Apoptotic; Arterial Fatty Streak; Atherosclerosis; BCL1 Oncogene; BCL2L1 gene; Biological Aging; Biology of Aging; Blood Vessels; CCL2 gene; Cardiac Myocytes; Cardiovascular Diseases; Cell Aging; Cell model; Cell physiology; Cells; Cellular biology; Cessation of life; Complement Factor B; Cultured Cells; DNA Sequence Alteration; Disease; Effectiveness; Endothelial Cells; Endothelium; Experimental Models; Functional disorder; GKLF protein; Gene Expression Regulation; Genes; Guanine; Guanosine; Heart Hypertrophy; Hydroxylation; Impairment; In Vitro; Inflammatory; Intercellular Adhesion Molecules; Interleukin-1 beta; Knock-out; Kruppel-like transcription factors; Lead; Ligase; Low Density Lipoprotein Receptor; Lung; Mesenchymal; Messenger RNA; MicroRNAs; Modification; Molecular; Morbidity - disease rate; Mus; Oxidative Stress; Patients; Phenotype; Pilot Projects; Population; Process; Proteins; Regulation; Reporting; Research Proposals; Rodent; Role; Specificity; Testing; Tissues; Transcript; Transforming Growth Factors; Tunica Intima; Ubiquitin; Untranslated RNA; Uracil; Vascular Cell Adhesion Molecule-1; aged; atherogenesis; atheroprotective; cardiovascular risk factor; connective tissue growth factor; epigenetic regulation; epitranscriptome; epitranscriptomics; in vivo; mortality; mouse model; oxidized low density lipoprotein; protein B; senescence; transcriptome; transcriptome sequencing

Project Summary/Abstract Atherosclerosis is a sequela of aging, in part, because aging exacerbates the atherogenic factors (e.g., oxidative stress), which induces endothelial cells (ECs) senescence. MicroRNAs (miRs) are ubiquitously involved in epigenetic regulation of gene expression by targeting the 3’-untranslated region (3’UTR) of the protein-encoded transcripts. Recently, we have reported that the miR-483 endows anti-endoMT, anti-fibrotic, and anti- inflammatory effects on the endothelium, contributing to a homeostatic endothelium. Emerging evidence indicates that oxidative stress can cause hydroxylation of guanine (G) in miRs to generate 8-oxo-7,8-dihydro guanosine (8OH-G) which induces the guanine-to-uracil (G-U) transversion of miRs. Because 8OH-G pairs with adenine (A), the oxidative stress-modified miRs no longer target their authentic mRNA effectually. This epitranscriptomic change results in distinctive functions in the cell, therefore sways the disease process. In the newly conducted pilot studies, we found elevated levels of 8OH-G miR-483 in senescent ECs in culture and vascular intima in aging mice. Moreover, EC function is affected by the molar ratio of miR-483/8OH-G miR-483, revealed by RNA-seq. These newly acquired results lead to the overarching hypothesis that the aging-induced miR-483 to 8OH-G miR-483 transversion causes EC senescence and dysfunction. Mechanistically, this miR epitranscriptomic change alters mRNA targetomes in ECs, which instigates aging-induced EC dysfunction and thereby aggravating atherosclerosis. To test this hypothesis, the following two specific aims are proposed: Aim 1. To delineate the molecular mechanism by which aging-induced 8OH-G miR-483 impairs EC function. Specifically, we will decipher the 8OH-G miR-483 epitranscriptomes by investigating the aging-associated G-to- U transversion of miR-483 and the specificity of 8OH-G-miR-483 targeting mRNAs. We will also study the sequelae of 8OH-G miR-483 with respect to EC senescence. Aim 2. To investigate the atheroprone effect of 8OH-G miR-483 in aging mice. We will introduce atherosclerosis in young and aged iEC-miR-483-Tg and their wildtype littermate mice. Atherosclerosis and vascular functions will be compared among young and old mice as functions of G-to-8OH-G miR-483 ratio and deduced 8OH-G miR-483 regulatory network. With the use of in vitro and in vivo experimental models, this exploratory research proposal will decipher the role of miR-483 to 8OH-G miR-483 transversion in EC biology and aging-deteriorated atherosclerosis.

项目概要/摘要 动脉粥样硬化是衰老的后遗症，部分原因是衰老使致动脉粥样硬化的因素（例如氧化应激）恶化。 应激），它会诱导内皮细胞（EC）衰老。MicroRNA（miR）普遍参与其中。 通过靶向蛋白质编码的 3'-非翻译区 (3'UTR) 对基因表达进行表观遗传调控 最近，我们报道了 miR-483 具有抗 endoMT、抗纤维化和抗 - 炎症对内皮的影响，有助于内皮的稳态。 表明氧化应激可导致 miR 中的鸟嘌呤 (G) 羟基化生成 8-oxo-7,8-dihydro 鸟苷 (8OH-G) 可诱导 miR 的鸟嘌呤至尿嘧啶 (G-U) 转换，因为 8OH-G 与 miR 配对。 腺嘌呤 (A)，氧化应激修饰的 miR 不再有效地靶向其真实的 mRNA。 表观转录组的变化导致细胞中独特的功能，从而影响疾病的进程。 在新进行的试点研究中，我们发现培养物中的衰老 EC 中 8OH-G miR-483 水平升高， 此外，miR-483/8OH-G miR-483 的摩尔比影响 EC 功能。 RNA-seq 揭示了这些新获得的结果得出了一个总体假设：衰老引起的。 miR-483 到 8OH-G miR-483 的颠换会导致 EC 衰老和功能障碍。 表观转录组的变化改变了 EC 中的 mRNA 靶标，从而引发衰老引起的 EC 功能障碍， 为了检验这一假设，提出了以下两个具体目标： 1. 阐明衰老诱导的 8OH-G miR-483 损害 EC 功能的分子机制。 具体来说，我们将通过研究与衰老相关的 G-to- 来破译 8OH-G miR-483 表观转录组。 我们还将研究 miR-483 的 U 转换和 8OH-G-miR-483 靶向 mRNA 的特异性。 8OH-G miR-483 与 EC 衰老相关的后遗症 目的 2. 研究 8OH-G miR-483 的动脉粥样硬化效应。 衰老小鼠中的 8OH-G miR-483 我们将介绍年轻和老年 iEC-miR-483-Tg 及其它们的动脉粥样硬化。 野生型同窝小鼠的动脉粥样硬化和血管功能将在年轻和年老小鼠之间进行比较。 G-与 8OH-G miR-483 比率的功能，并利用体外推断 8OH-G miR-483 调控网络。 和体内实验模型，这项探索性研究计划将破译 miR-483 对 8OH-G 的作用 EC 生物学和衰老恶化的动脉粥样硬化中的 miR-483 颠倒。