Determining the Platelet Signaling Pathway(s) Critical in Venous Thrombosis Pathogenesis" it might change in the process of editing

项目摘要 静脉血栓形成（VT）每年影响近一百万美国人。其中一个主要的诱因 室性心动过速的主要原因是血流受限，导致血管炎症和血栓形成 阵静脉血栓含有血小板，血小板计数的变化与VT相关 人类和小鼠的结果。然而，很少有人知道的分子机制， 这些细胞导致静脉血栓形成。我这个项目的目标是 系统研究血小板信号通路如何促进血小板/白细胞 粘附、血栓生长和小鼠血管狭窄后血栓固结。我 主要假设是静脉血栓形成过程中所需的血小板信号通路 类似于在炎症部位确保血管完整性所需的那些。具体地说， 我的研究旨在描述血小板G蛋白偶联受体（GPCRs）， 基于酪氨酸的免疫受体激活基序（ITAM）受体和Rap 1-talin 1-整联蛋白 信号传导轴参与小鼠VT的发展。与我的主要假设一致，我 预期ITAM信号和talin 1对静脉血栓形成比GPCR更重要 Rap 1以下转基因小鼠系具有血小板整合素缺陷的记录 将使用信号传导：CLEC 2（Clec 2fl/flpf 4-Cre+）、Rap 1（Rap 1afl/flRap 1bfl/flpf 4-Cre+）和 塔林1（Tln 1fl/flpf 4-Cre+）。胶原蛋白受体、GPVI、ITAM的药理学抑制 也将使用信号分子、布鲁顿酪氨酸激酶（Btk）以及β 1和β 3整联蛋白。在 在体内，VT将通过部分结扎下腔静脉（IVC狭窄）诱导，金 用于小鼠VT研究的标准模型。在48小时的血流限制后处死小鼠， 收获血栓并称重。为了研究血小板在室性心动过速发生中的作用， 在结扎后2至3小时对IVC进行活体成像，并对血小板和白细胞进行定量 粘附于IVC壁。全血凝块收缩和对溶解的敏感性研究将 提供了血栓形成过程中特定血小板信号通路作用的重要信息 巩固和解决。成功完成拟议的研究将阐明 血小板信号通路，有助于静脉血栓的启动、传播和稳定。 血栓这一信息可能导致新的血小板为基础的治疗目标的识别 用于预防/治疗VT。

Project summary Venous thrombosis (VT) affects close to a million Americans annually. One of the main triggers of VT is flow restriction, which leads to vascular inflammation and initiation of thrombus formation. Venous thrombi contain platelets and changes in platelet count correlate with VT outcome in humans and mice. However, little is known about the molecular mechanisms by which these cells contribute to venous thrombogenesis. The goal of my project is to systematically investigate on how major platelet signaling pathways facilitate platelet/leukocyte adhesion, thrombus growth, and thrombus consolidation following vascular stenosis in mice. My main hypothesis is that the platelet signaling pathways required during venous thrombogenesis are similar to those required for securing vascular integrity at sites of inflammation. Specifically, my study aims to delineate if and how platelet G-protein-coupled receptors (GPCRs), immunoreceptor tyrosine-based activation motif (ITAM) receptors, and the Rap1-talin1-integrin signaling axis contribute to VT development in mice. Consistent with my main hypothesis, I expect ITAM signaling and talin1 to be more important for venous thrombogenesis than GPCRs and Rap1. The following transgenic mouse lines with documented defects in platelet integrin signaling will be used: CLEC2 (Clec2fl/flpf4-Cre+), Rap1 (Rap1afl/flRap1bfl/flpf4-Cre+) and Talin1 (Tln1fl/flpf4-Cre+). Pharmacological inhibition of the collagen receptor, GPVI, the ITAM signaling molecule, Bruton’s tyrosine kinase (Btk), and 1 and 3 integrins will also be used. In vivo, VT will be induced by partially ligating the inferior vena cava (IVC stenosis), the gold standard model for VT studies in mice. Mice will be sacrificed after 48 hrs of flow restriction and thrombi harvested and weighed. To investigate the role of platelets in the initiation of VT, I will use intravital imaging of the IVC 2 to 3 hours after ligation and quantify platelet and leukocyte adhesion to the IVC wall. Whole blood clot contraction and susceptibility to lysis studies will provide important information on the role of specific platelet signaling pathways during thrombus consolidation and resolution. Successful completion of the proposed studies will elucidate platelet signaling pathways that contribute to the initiation, propagation, and stability of venous thrombi. This information may lead to the identification of new platelet-based therapeutic targets for the prevention/treatment of VT.