Targeting Platelet Endocytosis and Exocytosis to Control Thrombosis

靶向血小板胞吞作用和胞吐作用来控制血栓形成

• 批准号: 10046272
• 负责人: SIDNEY Waldo WHITEHEART
• 金额: --
• 依托单位: VA MEDICAL CENTER - LEXINGTON, KY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-10-01 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046272
• 关键词: Acute; Address; Adhesions; Affect; Age; Aging; Amino Acid Sequence; Autophagocytosis; Biochemical; Biological Assay; Blood Platelets; Blood Vessels; Cardiovascular Diseases; Caring; Carotid Artery Injuries; Cause of Death; Cell physiology; Cellular biology; Cessation of life; Clot retraction; Coagulation Process; Communicable Diseases; Complex; Cytoplasmic Granules; Data; Disease; Endocytosis; Equilibrium; Event; Exocytosis; Female; Fibrinogen; Gene Deletion; Goals; Growth; Guanosine Triphosphate Phosphohydrolases; Health; Hemorrhage; Hemostatic function; Hyperactivity; In Vitro; Infarction; Integrins; Knock-out; Knockout Mice; Knowledge; Mediating; Membrane; Membrane Fusion; Mission; Modeling; Molecular; Molecular Chaperones; Molecular Probes; Morbidity - disease rate; Mouse Strains; Mus; Myocardial Infarction; Pathology; Patients; Physicians; Precision Medicine Initiative; Process; Protein Biochemistry; Proteins; Publishing; RNA Splicing; Reaction; Risk; Risk Factors; Role; S-nitro-N-acetylpenicillamine; SNAP receptor; Sampling; Stroke; Tail; Therapeutic Intervention; Thrombosis; Thrombus; Transgenic Mice; Translations; Veterans; Work; base; clinically significant; experimental study; familial hemophagocytic lymphohistiocytosis; genome wide association study; glycosylation; granuphilin; health management; improved; in vitro Assay; in vivo; insight; male; military veteran; mortality; platelet function; protein protein interaction; recruit; response; restenosis; risk variant; synaptotagmin; syntaxin; syntaxin 11; syntaxin A; syntaxin-2; target SNARE proteins; targeted treatment; therapeutic target; thrombotic; trafficking; treatment strategy; vesicular SNARE proteins

World-wide, spurious thrombosis accounts for 1 of 4 non-communicable disease deaths. Cardiovascular disease is a leading killer of aging US veterans and is the major cause of death in older female veterans. Understanding how to modulate thrombosis will significantly aide the VA's mission to improve veterans' health. Normally platelets respond to vascular damage and secrete granule cargo that are essential for recruiting more platelets and for generating a thrombus. This releasate promotes normal sequelae but can also contribute to occlusive pathologies such as strokes and heart attacks. In platelets, VAMPs, SNAP-23, and Syntaxin-11 form a membrane-spanning complex that mediates exocytosis. Formation of this complex requires a host of SNARE-regulators, e.g., Munc18b, STXBP5/tomosyn-1, and granuphilin/SLP4; however, the mechanisms by which these proteins control the complexity of the platelet release reaction (its rate, extent, and content) is uncertain. Platelets are also capable of other cellular processes (i.e., RNA splicing, translation, glycosylation, autophagy); however, their effects on platelet function are still unknown. Our data suggest that endocytosis affects thrombus growth by modulating platelet spreading and platelet-platelet contacts. Our goal is to manipulate the membrane trafficking in platelets, both endocytosis and exocytosis, in order to modulate occlusive thrombosis with only modest effects on hemostasis. To reach this goal, we must probe the molecular mechanisms of exocytosis. Our specific focus will be on Syntaxin-11 regulators. We will also use an endocytosis defective mouse strain to define the roles of endocytosis in thrombosis and hemostasis. Two aims are proposed: 1) Define the network of protein-protein interactions that affect Syntaxin-11-mediated membrane fusion and granule cargo release; and 2) Determine the roles of platelet endocytosis in hemostasis. To complete these aims, we will employ biochemical assays to define the interactions between the SNARE regulators and in vitro and in vivo functional assays, using transgenic mice, to define the roles of the specific Syntaxin-11 regulators and of endocytosis in thrombosis and hemostasis. Our results will expand the understanding of the molecular requirements and the sequence of protein-protein interactions controlling platelet exocytosis. We will also expand the mechanistic understanding of what cellular processes platelets can perform (i.e., endocytosis) and why they are important. Our results will be significant to the field since they will provide the needed mechanistic insights to identify potential targets for therapeutic intervention and to evaluate the relevance of the increasing volume of gene/risk associations that are guiding patient treatment strategies. This will enhance the anti-thrombotic treatment options available for the care of aging veterans.

在世界范围内，假性血栓形成占 4 例非传染性疾病死亡中的 1 例。 心血管疾病是美国老年退伍军人的主要杀手，也是老年人死亡的主要原因 女退伍军人。了解如何调节血栓形成将极大地帮助 VA 的使命 改善退伍军人的健康。通常血小板会对血管损伤做出反应并分泌颗粒物质 这对于招募更多血小板和形成血栓至关重要。此次发布宣扬 正常的后遗症，但也可能导致闭塞性疾病，如中风和心脏病发作。在 血小板、VAMP、SNAP-23 和 Syntaxin-11 形成跨膜复合物，介导 胞吐作用。该复合物的形成需要大量 SNARE 调节因子，例如 Munc18b、 STXBP5/tomosyn-1 和 Granuphilin/SLP4；然而，这些蛋白质控制的机制 血小板释放反应的复杂性（其速率、程度和内容）是不确定的。血小板是 还能够进行其他细胞过程（即RNA剪接、翻译、糖基化、自噬）； 然而，它们对血小板功能的影响仍不清楚。我们的数据表明内吞作用影响 通过调节血小板扩散和血小板与血小板接触来抑制血栓生长。我们的目标是 操纵血小板的膜运输，包括胞吞作用和胞吐作用，以便 调节闭塞性血栓形成，对止血作用有限。为了实现这一目标，我们 必须探究胞吐作用的分子机制。我们的具体重点是 Syntaxin-11 监管机构。我们还将使用内吞作用缺陷小鼠品系来定义内吞作用在 血栓形成和止血。提出了两个目标：1）定义蛋白质-蛋白质网络 影响 Syntaxin-11 介导的膜融合和颗粒货物释放的相互作用；和 2)确定血小板内吞作用在止血中的作用。为了完成这些目标，我们将 采用生化测定来定义 SNARE 调节剂与体外和体外之间的相互作用 使用转基因小鼠进行体内功能测定，以确定特定 Syntaxin-11 的作用 血栓形成和止血中的调节剂和内吞作用。我们的结果将扩大理解 控制血小板的分子要求和蛋白质-蛋白质相互作用的顺序 胞吐作用。我们还将扩大对血小板可以做什么的细胞过程的机制理解 执行（即内吞作用）以及它们为何重要。我们的成果对该领域具有重要意义，因为 他们将提供所需的机制见解，以确定治疗干预的潜在目标 并评估指导患者的不断增加的基因/风险关联的相关性 治疗策略。这将增强可用于护理的抗血栓治疗选择 年迈的退伍军人。