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

确定静脉血栓形成发病机制中至关重要的血小板信号通路”，在编辑过程中可能会发生变化

• 批准号: 10066469
• 负责人: Jean Marie Niyitegeka Mwiza
• 金额: $ 3.66万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10066469
• 关键词: Adhesions; Adhesives; Affect; Agammaglobulinaemia tyrosine kinase; Age; American; Animals; Biological Assay; Blood Cells; Blood Platelets; Blood Vessels; Blood coagulation; Cardiovascular system; Cell Adhesion; Cells; Cessation of life; Clinical Trials; Coagulation Process; Collagen Receptors; Cytolysis; Defect; Development; Disease; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Goals; Gold; Growth; Harvest; Hemostatic function; Hour; Human; ITAM; Impairment; In Vitro; Incidence; Inferior vena cava structure; Inflammation; Inflammatory; Injury; Integrins; Lead; Leg; Leukocytes; Life; Ligation; Maintenance; Mechanics; Megakaryocytes; Modeling; Molecular; Monoclonal Antibodies; Mus; Myocardial Infarction; Outcome; Pathogenesis; Pharmacology; Phase; Platelet Activation; Platelet Count measurement; Platelet aggregation; Play; Predisposition; Prevention; Process; Proteins; Pulmonary Embolism; RAP1A gene; RAS Superfamily Proteins; Resolution; Role; Secure; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Standard Model; Stenosis; Stroke; Talin; Testing; Thrombocytopenia; Thrombosis; Thrombus; Transgenic Mice; Veins; Venous; Venous Thrombosis; Weight; Whole Blood; atherosclerotic plaque rupture; base; combat; in vivo; intravital imaging; intravital microscopy; new therapeutic target; prevent; receptor; therapeutic target; thrombogenesis; thromboinflammation; vascular inflammation; venous thromboembolism

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.

项目概要 静脉血栓（VT）每年影响近百万美国人。主要触发因素之一 VT 的本质是血流限制，导致血管炎症和血栓形成 形成。静脉血栓含有血小板，血小板计数的变化与VT相关 在人类和小鼠中的结果。然而，人们对其分子机制知之甚少。 这些细胞有助于静脉血栓形成。我的项目的目标是 系统地研究主要血小板信号通路如何促进血小板/白细胞 小鼠血管狭窄后的粘附、血栓生长和血栓巩固。我的 主要假设是静脉血栓形成过程中所需的血小板信号通路 类似于确保炎症部位血管完整性所需的那些。具体来说， 我的研究旨在阐明血小板 G 蛋白偶联受体 (GPCR) 是否以及如何， 基于免疫受体酪氨酸的激活基序 (ITAM) 受体和 Rap1-talin1-整合素 信号轴有助于小鼠 VT 的发展。与我的主要假设一致，我 预计 ITAM 信号传导和 talin1 对于静脉血栓形成比 GPCR 更重要 和说唱1。以下转基因小鼠品系已记录有血小板整合素缺陷 将使用的信号：CLEC2 (Clec2fl/flpf4-Cre+)、Rap1 (Rap1afl/flRap1bfl/flpf4-Cre+) 和 Talin1 (Tln1fl/flpf4-Cre+)。胶原蛋白受体、GPVI、ITAM 的药理抑制 还将使用信号分子、布鲁顿酪氨酸激酶 (Btk) 以及 1 和 3 整合素。在 体内，VT将通过部分结扎下腔静脉（IVC狭窄）来诱导，金 小鼠 VT 研究的标准模型。限流 48 小时后处死小鼠， 收获血栓并称重。为了研究血小板在 VT 启动中的作用，我将 结扎后 2 至 3 小时使用 IVC 活体成像并量化血小板和白细胞 粘附于 IVC 壁。全血凝块收缩和对溶解的敏感性研究将 提供有关血栓形成过程中特定血小板信号传导途径作用的重要信息 巩固和解决。成功完成拟议的研究将阐明 血小板信号传导途径有助于静脉血流的启动、传播和稳定性 血栓。该信息可能有助于识别新的基于血小板的治疗靶点 用于预防/治疗 VT。