Mechanisms of Platelet Activity in Vascular Disease

项目摘要/摘要 据估计，美国有1500万至2000万人患有外周动脉疾病(PAD)。尽管 医学治疗的进展，PAD仍然与相当大的心脏和四肢发病率和 死亡率。目前，在下肢进行的侵入性手术比在心脏进行的更多， 证明了先进的PAD系统的成本不断增加。虽然冠状动脉的发病机制 冠心病(CAD)的特点很好，对PAD的病理生理和发病机制了解较少(S) 对这一复杂疾病的监管仍不确定。而抗血小板治疗(作为一种阶级效应)减少 PAD的发生率和并发症，我们注意到抗血小板治疗的有效性不同。 PAD和其他血管表型之间的差异。与冠心病相比，阿司匹林对PAD并不是特别有效 更有效的抑制P2Y12也没有临床上的好处。显然，需要新的方向来更好地 了解血小板在PAD发病机制中的作用并确定新的治疗靶点。我们的团队 证明了编码和非编码RNA在调节血小板活性中的重要性。利用我们的 建立了具有良好表型血小板活性的PAD患者队列，我们证明了 PAD发病机制中的血小板-白细胞相互作用(与血小板-血小板聚集相反)。 此外，我们发现了PAD中血小板转录后的异常调节，并证明了 在PAD中，血小板在激活单核细胞和促进炎症中起着中心效应作用。在这里，我们 建议综合研究(1)血小板活性、(2)血小板 转录组，以及(3)PAD患者的效应细胞特性。我们会分析储存的血小板样本 来自&gt；1000名患者进行了纵向随访，其中许多人提供了系列收集。利用这些优势 有价值的血小板样本，我们将专注于鉴定与血管相关的新的血小板转录本 表型和偶发心血管事件。例如，我们将比较(1)PAD患者与其他患者 血管表型(例如，冠心病、颈动脉狭窄、腹主动脉瘤)，(2)稳定垫与CLI， (3)心脏(心肌梗死)与肢体(主要截肢)事件。机械学研究使用 培养的巨核细胞和具有血小板特异性的候选基因敲除和敲除的动物模型 基因将描述这些过程是如何被调控的。我们也处于有利地位，可以验证我们的发现 在成熟的地方、国家和国际队列中。我们的数据表明这些类型的研究可以 提供概念上的进展，使我们了解影响发病机制和 PAD的严重程度。这些见解可以被用来设计临床生物标记物和治疗策略 治疗和预防血管疾病及其危及生命的并发症。

PROJECT SUMMARY/ABSTRACT An estimated 15–20 million people in the United States have peripheral artery disease (PAD). Despite advances in medical therapy, PAD remains associated with considerable cardiac and limb morbidity and mortality. Currently, more invasive procedures are performed in the lower extremities than in the heart, demonstrating increasing costs to the system of advanced PAD. Although the pathogenesis of coronary artery disease (CAD) is well characterized, the pathophysiology of PAD is less understood and the mechanism(s) that regulate this complex disorder remain uncertain. While antiplatelet therapy (as a class effect) decreases the incidence and complications from PAD, we noted that the effectiveness of antiplatelet therapy differs between PAD and other vascular phenotypes. In contrast to CAD, aspirin was not particularly effective in PAD nor was there clinical benefit with more potent P2Y12 inhibition. Clearly, new directions are needed to better understand the role of platelets in PAD pathogenesis and identify new therapeutic targets. Our group demonstrated the importance in coding and noncoding RNAs in regulating platelet activity. Leveraging our established cohort of PAD patients with well-phenotyped platelet activities, we demonstrated the importance of platelet–leukocyte interactions (in contrast to platelet–platelet aggregation) in the pathogenesis of PAD. Moreover, we identified an aberrant post-transcriptional regulation of platelets in PAD and demonstrated that platelets play a central effector role in activating monocytes and fostering inflammation in PAD. Here, we propose to comprehensively investigate the relationship between (1) platelet activity, (2) the platelet transcriptome, and (3) effector cell properties in patients with PAD. We will analyze stored platelet samples from >1,000 patients with longitudinal follow-up, many of whom provided serial collections. Leveraging these valuable platelet samples, we will focus on identifying novel platelet transcripts associated with vascular phenotypes and incident cardiovascular events. For example, we will compare patients with (1) PAD vs. other vascular phenotypes (e.g., CAD, carotid artery stenosis, abdominal aortic aneurysm), (2) stable PAD vs. CLI, and (3) incident cardiac (myocardial infarction) vs. limb (major amputation) events. Mechanistic studies using both cultured megakaryocytes and animal models with platelet-specific knock-in and knock-out of candidate genes will characterize how these processes are regulated. We are also well positioned to validate our findings in well-established local, national, and international cohorts. Our data suggest these types of studies can provide conceptual advances in our understanding of the mechanisms influencing the pathogenesis and severity of PAD. These insights could be leveraged to design clinical biomarkers and therapeutic strategies to treat and prevent vascular disease and its life-threatening complications.