Resubmission: Structure/Function Determinants of Puncture Wound Thrombus Formation

重新提交：刺伤血栓形成的结构/功能决定因素

• 批准号: 10625508
• 负责人: Brian Storrie
• 金额: $ 64.03万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10625508
• 关键词: 3-Dimensional; Accidents; Achievement; Adhesives; Affect; Alpha Granule; Antiplatelet Drugs; Arteries; Aspirin; Autocrine Communication; Blood Coagulation Factor; Blood Platelets; Blood Vessels; Cardiovascular system; Cartoons; Coagulation Process; Collagen Receptors; Complex; Cytoplasmic Granules; Data; Defect; Deposition; Diameter; Disease susceptibility; Dose; Drug Modulation; Drug Targeting; Embolism; Event; Experimental Models; Fibrin; Fibrinogen Receptors; Fibrinolytic Agents; Future; Goals; Growth; Health; Hemorrhage; Hemostatic function; Hermanski-Pudlak Syndrome; Human; Individual; Knowledge; Life; Light; Maintenance; Medical; Medicine; Microscopy; Modeling; Molecular; Morphology; Mus; Operative Surgical Procedures; Oranges; Outcome; Patients; Pattern; Penetration; Pharmaceutical Preparations; Pharmacotherapy; Physiological; Platelet Activation; Platelet aggregation; Predictive Value; Probability; Process; Puncture biopsy; Puncture procedure; Puncture wound; Reproducibility; Research; Resources; Risk; Role; SNAP receptor; Signal Transduction; Site; Stratification; Structure; Structure of jugular vein; Syndrome; System; Testing; Thrombin; Thrombosis; Thromboxanes; Thrombus; Time; Traumatic injury; VAMP-2; Vascular System; Venous; Visualization; Wound models; adhesion receptor; arteriole; autocrine; clopidogrel; comparative; driving force; drug action; experimental study; genetic manipulation; hazard; inhibitor; meter; model building; mouse genetics; mutant; novel; predictive modeling; public health relevance; receptor; response; side effect; traumatic event; venule; von Willebrand factor receptor; wound

PROJECT SUMMARY/ABSTRACT Scientific Premise and Prior Research: Puncture wounds, be they produced by accident, intent or medical practice, e.g., surgery, can be a routine to major traumatic event. Moreover, drug aggravated bleeding is a major limitation of current antiplatelet drugs that form a cornerstone of cardiovascular patient therapy and a potentially major liability to any surgery. We contend that detailed structure understanding of the platelet centric responses that occur within the actual puncture hole is essential to fill a void in prior research and yield an important framework for the management traumatic injuries and common bleeding side effects. Gap(s) in knowledge: Previous approaches to thrombus formation have focused mostly on examples in which the exposed vascular matrix remains intact and therefore little to no bleeding occurs and have suffered from the inability of 2P microscopy or SEM to assess platelet activation at the level of individual platelets. From this circumscribed experimental perspective, a “Core and Shell” model of thrombus structure, i.e., a horizontal core of matrix anchored, highly activated platelets covered by a shell of weakly activated platelets in which signaling within the Core is thrombin-dependent and within the Shell is ADP/thromboxane-dependent has emerged. We submit that this model may well be limited in its application to actual bleeding where the wound is an open puncture hole. Solution to problem: Taking alternate visualization approaches that determine the activation state of individual platelets in 3D space, our Preliminary Studies have already yielded data redefining normal, bleeding cessation as exemplified in a puncture wound model. Our data reveal a spatially dispersed, platelet activation pattern and strong indications that initial steps in thrombus formation are dependent on the capture of platelet aggregates that cap the hole from the outside rather than fill the hole. This has led us to formulate a new “Cap and Build” paradigm. How both platelet-centric and coagulation factor cascade events can be integrated within the paradigm is now an important question. To understand these multiple inputs through experimentation, we have proposed Aims that focus first on signaling events, be they autocrine (platelet secretion, Specific Aim 1) or exocrine (as inferred from the effects of anti-platelet drugs or DOACs, Specific Aim 2), and the importance of platelet adhesion receptors (Specific Aim 3). We also propose to test how well the Cap and Build paradigm apply to smaller, less traumatic injury.

项目摘要/摘要 科学前提和先前的研究：穿刺伤口，无论是偶然，意图还是医疗产生 练习，例如，手术可能是重大创伤事件的常规。而且，药物汇总出血是主要的 构成心血管疗法的基石的当前抗血小板药物的局限性 任何手术的重大责任。我们认为对血小板中心反应的详细结构理解 在实际的穿刺孔内发生的，对于在先前的研究中填充空隙至关重要，并产生重要的 管理创伤性损伤和常见出血副作用的框架。 知识的差距：先前的血栓形成方法主要集中在示例上 暴露的血管基质保持完好无损，因此几乎没有出血，并且遭受了 从2p显微镜或SEM的无能为力，以评估单个血小板水平的血小板激活。 从这个限制的实验角度来看，血栓结构的“核心和外壳”模型，即 基质的水平核心锚定，高度激活的血小板，被弱激活血小板的外壳覆盖 核心内的哪个信号取决于凝血酶依赖性，并且壳内是ADP/Thromboxane依赖性的 出现了。我们认为，该模型的应用可能会限制在实际出血中 伤口是一个开放的穿刺孔。 解决问题：采用确定个体激活状态的替代可视化方法 在3D空间中的血小板，我们的初步研究已经产生了数据重新定义正常，出血停止 如穿刺伤口模型所示。我们的数据揭示了空间分散的，血小板激活模式和 有力的迹象表明，血栓形成的初始步骤取决于血小板聚集体的捕获 那个孔从外部限制而不是填充孔。这使我们制定了新的“帽子和构建” 范例。如何将以血小板为中心和凝结因子级联事件集成 范式现在是一个重要的问题。为了通过实验了解这些多个输入，我们有 提出的目标是首先关注信号事件的目标，无论是自动分泌（血小板分泌，特定目的1）还是 外分泌（根据抗血小板药物或DOAC的影响，特定目标2），以及重要性 血小板粘附受体（特定目标3）。我们还建议测试盖帽和构建范式的效果 适用于较小的，减少创伤性的伤害。

项目成果

Tethered platelet capture provides a mechanism for restricting circulating platelet activation to the wound site.

• DOI: 10.1016/j.rpth.2023.100058
• 发表时间: 2023-02
• 期刊: RESEARCH AND PRACTICE IN THROMBOSIS AND HAEMOSTASIS
• 影响因子: 4.6
• 作者: Pokrovskaya, Irina D.;Rhee, Sung W.;Ball, Kelly K.;Kamykowski, Jeffrey A.;Zhao, Oliver S.;Cruz, Denzel R. D.;Cohen, Joshua;Aronova, Maria A.;Leapman, Richard D.;Storrie, Brian
• 通讯作者: Storrie, Brian