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.

项目概要/摘要 科学前提和先前研究：刺伤，无论是意外、故意还是医疗造成的 实践，例如手术，可能是重大创伤事件的常规，而且药物加重出血是主要的。 当前抗血小板药物的局限性，这些药物构成了心血管患者治疗的基石，并且具有潜在的潜力 我们认为对血小板中心反应的详细结构理解是任何手术的主要责任。 发生在实际穿刺孔内的现象对于填补先前研究的空白至关重要，并产生重要的结果 处理创伤性损伤和常见出血副作用的框架。 知识差距：以前的血栓形成方法主要集中在以下例子 暴露的血管基质保持完整，因此几乎没有出血或没有出血 2P 显微镜或 SEM 无法在个体血小板水平评估血小板活化。 从这个有限的实验角度来看，血栓结构的“核和壳”模型，即 基质锚定的水平核心，高度活化的血小板，被弱活化的血小板外壳覆盖 核心内的信号传导依赖于凝血酶，而壳内的信号传导依赖于 ADP/血栓烷 我们认为该模型的应用很可能仅限于实际出血。 伤口是一个开放的穿刺孔。 问题的解决方案：采用替代可视化方法来确定个体的激活状态 3D 空间中的血小板，我们的初步研究已经获得了重新定义正常出血停止的数据 正如穿刺伤模型所示，我们的数据揭示了空间分散的血小板激活模式和 强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 从外部堵住漏洞而不是填补漏洞 这促使我们制定了新的“上限和构建”。 以血小板为中心的事件和凝血因子级联事件如何整合到系统中。 为了通过实验理解这些多重输入，范式现在是一个重要的问题。 提出的目标首先关注信号事件，无论是自分泌（血小板分泌，具体目标 1）还是 外分泌（根据抗血小板药物或 DOAC 的作用推断，具体目标 2），以及外分泌的重要性 我们还建议测试 Cap 和 Build 范例的效果。 适用于较小的、较少的外伤。

项目成果

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

束缚血小板捕获提供了一种限制伤口部位循环血小板活化的机制。

• 发表时间: 2023-02
• 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

Ferric Chloride-Induced Arterial Thrombosis and Sample Collection for 3D Electron Microscopy Analysis.

氯化铁诱导的动脉血栓形成和样品采集用于 3D 电子显微镜分析。

• 发表时间: 2023-03-17
• 作者: Joshi, Smita;Smith, Alexis N;Prakhya, Kanakanagavalli Shravani;Alfar, Hammodah R;Lykins, Joshua;Zhang, Ming;Pokrovskaya, Irina;Aronova, Maria;Leapman, Richard D;Storrie, Brian;Whiteheart, Sidney W
• 通讯作者: Whiteheart, Sidney W