MicroRNA Profile in Peripheral Artery Disease

描述(申请人提供)：外周动脉疾病(PAD)日益普遍，并导致残疾和高心血管风险。临床状态的决定因素 对PAD的预后尚不清楚。来自实验模型的新证据表明，microRNAs(MiRNAs)是肢体缺血细胞反应的调节因子和潜在的治疗靶点。MiRNA表达改变可能通过调节血管生成、炎症和内皮损伤反应而参与PAD的发生，但对患者的这些机制知之甚少。MiRNAs在血液中循环，具有作为生物标志物的潜力，并可能作为内分泌信号因子发挥作用。内皮细胞是循环miRNA的潜在来源。我们已经开发出了基于全面定量RT-PCR的血浆中miRNA图谱的方法。此外，我们已经建立了在新鲜分离的内皮细胞中表征miRNA表达的能力。这项拟议的研究旨在定义PAD中循环和动脉内皮细胞的miRNAs，以深入了解PAD临床疾病的机制。我们将通过研究我们特征良好的、由国家心肺和血液研究所资助的PAD队列和对照组(306名PAD患者、50名CAD患者和50名对照组)进行纵向结果评估来实现目标1和目标2，并通过招募90名患者(每个PAD、单独的CAD和对照组各30名)进行内皮细胞分离来实现目标3。在目标1中，我们将通过比较PAD患者与冠心病患者和健康对照组来验证PAD改变循环miRNA表达的假设。在目标2中，我们将在PAD中将循环miRNAs与疾病严重程度、炎性细胞因子和循环血管生成调节因子联系起来。此外，我们将使用之前收集的两年随访数据来验证循环miRNAs与PAD中的冠状动脉和外周疾病事件相关的假设。在目标3中，我们将在血管造影术时通过J钢丝活检从髂股动脉收集动脉内皮细胞。我们将内皮细胞miRNA的表达与循环miRNAs和PAD状态联系起来。这些研究将提供有关PAD中循环和内皮miRNAs的新信息，并可能刺激创新的治疗策略。

DESCRIPTION (provided by applicant): Peripheral artery disease (PAD) is increasingly prevalent and leads to disability and high cardiovascular risk. The determinants of clinical status and prognosis in PAD are not well understood. Emerging evidence from experimental models identifies microRNAs (miRNAs) as regulators of cellular responses to limb ischemia and as potential therapeutic targets. Altered miRNA expression may contribute to PAD by modulating angiogenesis, inflammation and endothelial injury response, but little is known about these mechanisms in patients. MiRNAs circulate in the blood, hold potential as biomarkers, and may function as endocrine signaling factors. Endothelial cells are a potential source for circulating miRNA. We have developed methods to perform comprehensive quantitative RT-PCR based miRNA profiling in plasma. In addition, we have established the ability to characterize miRNA expression in freshly isolated endothelial cells. The proposed research aims to define circulating and arterial endothelial miRNAs in PAD to provide insight into the mechanisms driving clinical disease in PAD. We will accomplish Aims 1 and 2 by studying our well-characterized, National Heart Lung and Blood Institute-funded PAD cohort and controls (306 PAD patients, 50 CAD patients and 50 controls) with longitudinal outcome evaluation and Aim 3 by enrolling 90 patients (30 each PAD, CAD alone and control) for endothelial cell isolation. In Aim 1, we will test the hypothesis that PAD alters circulating miRNA expression by comparing PAD patients to CAD and healthy controls. In Aim 2, we will relate circulating miRNAs to disease severity, inflammatory cytokines, and circulating angiogenesis modulators in PAD. Further, we will use previously collected 2-year follow-up data to test the hypothesis that circulating miRNAs are associated with coronary and peripheral disease events in PAD. In Aim 3, we will collect arterial endothelial cells at the time of angiography by J-wire biopsy from the iliofemoral arteries. We relate endothelial cell miRNA expression to circulating miRNAs and PAD status. These studies will provide new information regarding circulating and endothelial miRNAs in PAD and may stimulate innovative treatment strategies.