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: 10540297
• 负责人: Jean Marie Niyitegeka Mwiza
• 金额: $ 2.94万
• 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 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10540297
• 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.

项目摘要 静脉血栓形成（VT）每年接近一百万美国人。主要触发器之一 VT的流量限制，这导致血管注射和血栓的主动性 形成。静脉血栓包含血小板和血小板计数的变化与VT相关 人类和小鼠的结果。但是，对分子机制的了解鲜为人知 这些细胞有助于静脉血栓形成。我项目的目标是 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期间需要的血小板信号通路 与在炎症部位获得血管完整性所需的相似。具体来说， 我的研究旨在描绘出以及血小板G蛋白偶联受体（GPCR），如何描述。 免疫受体酪氨酸的活化基序（ITAM）受体和RAP1-TALIN1-整合蛋白 信号轴有助于小鼠的VT发育。与我的主要假设一致 期望ITAM信号传导和talin1比GPCR对静脉血栓形成更为重要 和Rap1。以下具有血小板整合素中有记录缺陷的转基因鼠标线 将使用信号：CLEC2（CLEC2FL/FLPF4-CRE+），RAP1（RAP1AFL/FLRRAP1BFL/FLPF4-CRE+）和 talin1（tln1fl/flpf4-cre+）。胶原受体GPVI，ITAM的药理抑制 信号分子，布鲁顿的酪氨酸激酶（BTK）以及1和3整合素也将使用。 VIVO，VT将通过部分连接下腔静脉（IVC狭窄），金，金诱导。 小鼠VT研究的标准模型。 48小时的流动限制后，将牺牲小鼠 血栓收获并称重。为了调查血小板在VT计划中的作用，我将 连接后2至3小时，使用IVC的插入成像并量化血小板和白细胞 对IVC墙的粘附。全血凝块收缩和对裂解研究的敏感性将 提供有关血栓期间特定血小板信号通路作用的重要信息 合并和解决方案。成功完成拟议的研究将阐明 静脉的主动性，繁殖和稳定性的血小板信号通路 血栓。此信息可能导致识别新的基于血小板的治疗靶标 用于预防/治疗VT。