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: 10793303
• 负责人: Ming He
• 金额: $ 18.56万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10793303
• 关键词: 3' Untranslated Regions; Acceleration; Adenine; Adrenergic Agonists; Affect; Aging; Antiinflammatory Effect; Apoptotic; Arterial Fatty Streak; Atherosclerosis; BCL1 Oncogene; BCL2L1 gene; Biological Aging; Blood Vessels; CCL2 gene; Cardiac Myocytes; Cardiovascular Diseases; Cell Aging; Cell Separation; Cell model; Cell physiology; Cells; Cellular biology; Cessation of life; Cultured Cells; DNA Sequence Alteration; Deterioration; Disease; Effectiveness; Endothelial Cells; Endothelium; Endowment; Experimental Models; Fibrosis; Functional disorder; GKLF protein; Gene Expression Regulation; Genes; Guanine; Guanosine; Heart Hypertrophy; Hydroxylation; Impairment; In Vitro; Inflammation; Inflammatory; Intercellular Adhesion Molecules; Intercellular adhesion molecule 1; Interleukin-1 beta; Knockout Mice; Kruppel-like transcription factors; Ligase; Low Density Lipoprotein Receptor; Lung; Mesenchymal; Messenger RNA; MicroRNAs; Modification; Molecular; Morbidity - disease rate; Mus; Oxidative Stress; Patients; Phenotype; Pilot Projects; Population; Process; Protein Region; Regulation; Reporting; Research Proposals; Rodent; Role; Specificity; Testing; Tissues; Transcript; Transforming Growth Factor beta; 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; human old age (65+); 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.

项目摘要/摘要 动脉粥样硬化是衰老的后遗症，部分原因是衰老加剧了致动脉粥样硬化的因素(例如，氧化 应激)，诱导内皮细胞(ECs)衰老。MicroRNAs(MiRs)广泛参与 通过靶向蛋白质编码的3‘非翻译区对基因表达的表观遗传调控 成绩单。最近，我们报道了miR-483具有抗内毒素、抗纤维化和抗血管紧张素转换酶的作用。 炎症对内皮的影响，促进内皮细胞的动态平衡。新出现的证据 表明氧化应激可导致miRs中鸟嘌呤(G)的羟基化生成8-氧代-7，8-二氢 鸟苷(8OH-G)可诱导miRs鸟嘌呤向尿嘧啶(G-U)的转换。因为8OH-G与 腺嘌呤(A)，氧化应激修饰的miRs不再有效地靶向其真实的mRNA。这 表位转录的变化会导致细胞中不同的功能，从而影响疾病的进程。在 最新进行的先导研究发现，在培养和培养的衰老内皮细胞中，8OH-G miR-483水平升高 衰老小鼠的血管内膜。此外，miR-483/8OH-G miR-483的摩尔比对EC功能有影响， 由RNA-seq.这些新获得的结果导致了一个重要的假设，即衰老导致 MiR-483到8OH-G miR-483的颠倒导致EC衰老和功能障碍。从机械上讲，这款MIR 表位转录改变改变内皮细胞的mRNA靶体，从而引发衰老诱导的内皮细胞功能障碍和 从而加重动脉粥样硬化。为了验证这一假设，提出了以下两个具体目标： 1.探讨衰老诱导8OH-G miR-483损伤内皮细胞功能的分子机制。 具体地说，我们将通过研究与衰老相关的G-to-483来破译8OH-G miR-483表位转录。 MiR-483的U翻转及8OH-G-miR-483靶向mRNAs的特异性。我们亦会研究 8OH-G miR-483与EC衰老的后遗症。目的2.研究丹参的动脉粥样硬化效应。 8OH-G miR-483对衰老小鼠的影响。我们将介绍年轻人和老年人的动脉粥样硬化-IEC-miR-483-TG及其 野生型产仔小鼠。动脉粥样硬化和血管功能将在年轻和老年小鼠之间进行比较， G-to-8OH-G miR-483比值的作用，推导出8OH-G miR-483调控网络。通过使用体外培养技术 在活体实验模型中，这一探索性研究方案将破译miR-483到8OH-G的作用 MIR-483在EC生物学中的颠倒与衰老恶化的动脉粥样硬化。