miR-615, AKT/eNOS signaling, and angiogenesis

miR-615、AKT/eNOS 信号传导和血管生成

基本信息

批准号：
10594486
负责人：
MARK W FEINBERG
金额：
$ 62.95万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10594486
关键词：
3&apos Untranslated Regions AKT inhibition Acute Angiogenesis Inhibitors Base Pairing Binding Biological Assay Biology Blood Vessels Blood capillaries Blood flow Brain Cardiovascular Diseases Cardiovascular Physiology Cardiovascular system Cell Proliferation Cell physiology Cellular biology Chronic Clinical Research Disease Endothelial Cells Endothelium FGF2 gene Foundations Gene Expression Genes Goals Growth Growth Factor Health Expenditures Heart Hindlimb Human IGF2 gene Impairment Infarction Injections Ischemia Knockout Mice Leg Ligation Limb structure Link Mediating Mediator Messenger RNA MicroRNAs Molecular Morbidity - disease rate Mus Mutation Myocardial Myocardial Infarction Myocardial Ischemia Nitric Oxide Organoids Outcome Participant Pathway interactions Patients Peripheral arterial disease Phenocopy Plasma Process Proliferating Proto-Oncogene Proteins c-akt Publishing Recovery Regulation Reperfusion Therapy Reporter Genes Repression Resistance Role Signal Pathway Signal Repression Signal Transduction Stimulus Therapeutic Thrombospondin 1 Tissue Sample Tissues Tube United States Untranslated RNA Vascular Endothelial Cell Vascular Endothelial Growth Factors acute coronary syndrome angiogenesis artery occlusion blood vessel development cell growth cell motility crosslinking and immunoprecipitation sequencing gene repression human disease human subject human tissue improved in vivo inhibitor insight intravenous administration ischemic injury loss of function matrigel miRNA expression profiling migration molecular imaging mortality myocardial infarct sizing nanoparticle neovascularization novel novel therapeutic intervention novel therapeutics overexpression perfusion imaging posttranscriptional preclinical study response response to injury tissue repair transcriptome sequencing transcriptomic profiling wound closure

项目摘要

Ischemic cardiovascular disease (CVD) due to atherosclerotic occlusion of the arteries to the heart, legs, or brain is associated with considerable morbidity, mortality, and health care expenditure in the United States. The induction and orchestration of new blood vessels is critical for tissue repair in response to injury such as myocardial infarction or peripheral artery disease (PAD). In response to pro-angiogenic stimuli, vascular endothelial cells (ECs) are activated to migrate and proliferate to form primary capillaries. However, despite the importance of ECs in neoangiogenesis, our understanding of the mechanisms regulating this process remains poorly understood. Emerging studies indicate that the inability of angiogenic growth factors to stimulate angiogenesis is likely due to impaired angiogenic signaling and not due to deficiency in these growth factors. MicroRNAs (miRNAs) are small, single-stranded, non-coding RNAs capable of repressing gene expression by base pairing to the 3' untranslated regions (3'-UTRs) of mRNA targets and are involved in a variety of pathophysiological processes in cardiovascular biology, though their function in angiogenesis and angiogenic signaling pathways remains poorly defined. We undertook a microarray profiling approach of plasma from subjects with ischemic CVD and identified that miR-615-5p expression is increased by ischemia and reduced in response to pro-angiogenic stimuli–observations that are recapitulated in both mice and human ischemic paradigms in vivo. Preliminary and published gain and loss-of-function studies reveal that miR-615-5p overexpression markedly impaired EC proliferation, migration, and network tube formation in matrigel, whereas blockade of miR-615-5p had the opposite effects. Mechanistically, using unbiased transcriptomic profiling, we find that miR-615-5p suppressed EC proliferation and binding to 2 unique targets–RASSF2 and IGF2–in their 3'-UTRs and reduced their expression, an effect that selectively regulated the AKT/eNOS signaling pathway in ECs. Finally, systemic intravenous administration of miR-615-5p inhibitors increased blood vessel formation and reduced infarct size and improved blood flow recovery in ischemic legs compared to mice that received scrambled control anti-miR injections. These observations provide the foundation for the central hypothesis that miR-615-5p may serve as a critical regulator of EC proliferation and angiogenic responses. To better understand the precise role of miR-615-5p in AKT/eNOS signaling and angiogenesis, we will in Aim1 delineate the upstream mechanisms governing miR-615-5p expression in ECs. In Aim2, we will determine the molecular basis for miR-615-5p's ability to regulate AKT/eNOS signaling and EC functions critical to angiogenesis. In Aim3, we will explore the effect of altering miR-615-5p expression in the microvasculature on acute and chronic experimental ischemic injury. The results of these studies will provide insights regarding miR-615-5p function in EC biology, pathophysiological angiogenesis, and cardiovascular ischemic states and may provide new targets to rescue impaired angiogenic signaling for a range of ischemic cardiovascular disease states.

由于动脉的动脉粥样硬化，心脏，腿部或大脑与美国的考虑发病率，死亡率和医疗保健支出有关。新血管的诱导和编排对于响应损害的组织修复至关重要心肌梗塞或周围动脉疾病（PAD）。响应促血管生成刺激，血管内皮细胞（EC）被激活以迁移和增殖以形成原发性毛细血管。但是，dospite EC在新血管生成中的重要性，我们对调节此过程的机制的理解仍然存在理解不佳。新兴研究表明，血管生长因子无法刺激血管生成可能是由于血管生成信号受损而导致的，而不是由于这些生长因子缺乏。 microRNA（miRNA）是小的，单链的非编码RNA，能够反射基因通过基础配对与mRNA靶标的3'未翻译区（3'-UTR）表达，并参与了A 心血管生物学中的各种病理生理过程，尽管它们在血管生成和血管生成信号通路的定义较差。我们采用了一种微阵列分析方法缺血性CVD受试者的血浆，并确定缺血增加了miR-615-5p的表达并减少了在小鼠和人类中都概括的促血管生成刺激的观察体内缺血范式。初步和发表的功能丧失研究表明，miR-615-5p 过表达明显损害了矩阵中的EC增殖，迁移和网络管形成，而 miR-615-5p的封锁具有相反的影响。从机械上讲，使用公正的转录组分析，我们发现miR-615-5p抑制了EC的增殖并与2个唯一靶标-RASSF2和IGF2结合 - 3'-UTRS并降低了它们的表达 ECS。最后，全身静脉注射miR-615-5p抑制剂增加了血管的形成与接受的小鼠相比炒控制抗MIR注射。这些观察为中心假设奠定了基础 miR-615-5p可以作为EC增殖和血管生成反应的关键调节剂。更好了解miR-615-5p在Akt/eNOS信号传导和血管生成中的精确作用，我们将在AIM1中在EC中涉及miR-615-5p表达的上游机制。在AIM2中，我们将确定分子 miR-615-5p调节Akt/eNOS信号传导和EC功能对血管生成至关重要的能力的基础。在 AIM3，我们将探讨微脉管系统中改变miR-615-5p表达对急性和慢性实验性缺血性损伤。这些研究的结果将提供有关miR-615-5p的见解 EC生物学，病理生理血管生成和心血管缺血状态的功能，可能会提供为一系列缺血性心血管疾病状态挽救血管生成信号受损的新目标。