Determinants of Thrombus Structure and Stability

血栓结构和稳定性的决定因素

• 批准号: 8903582
• 负责人: Alisa S. Wolberg
• 金额: $ 37.78万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8903582
• 关键词: Address; Affect; American; Binding; Biochemical Genetics; Biological; Biological Assay; Biological Models; Biophysical Process; Blood; Cell physiology; Characteristics; Coagulation Process; Data; Deep Vein Thrombosis; Development; Employee Strikes; Enzymes; Erythrocytes; Factor XIII; Fibrin; Fibrinogen; Figs - dietary; Goals; Health; Hemostatic function; Human; Immunohistochemistry; In Vitro; Individual; Maps; Mediating; Medical; Microscopy; Mission; Modeling; Molecular; Molecular Target; Mus; Outcome; Patients; Pharmaceutical Preparations; Pharmacia brand of estropipate; Phase; Plasma; Population; Production; Public Health; Pulmonary Embolism; Recording of previous events; Recurrence; Research; Resolution; Risk; Role; Solutions; Structure; Thrombelastography; Thrombin; Thromboembolism; Thrombosis; Thrombus; Ultrasonography; Venous; Venous Thrombosis; Whole Blood; Work; analytical method; blocking factor; clinically significant; crosslink; human disease; in vivo; inhibitor/antagonist; innovation; intravital microscopy; knowledge base; novel; prevent; reconstitution; thrombolysis; tool

DESCRIPTION (provided by applicant): Venous thrombosis/thromboembolism affects over 1 million Americans per year. Thrombus burden (size) predicts long-term outcome. Efforts to reduce venous thrombus burden with the development of new drugs could have a considerable impact on the US population. The defining characteristic of venous "red" thrombi is their high red blood cell (RBC) content, which was traditionally thought to result from passive trapping of RBCs in the static fibrin clot. However, our new preliminary data indicate that RBCs are retained in clots via an active mechanism that requires the fibrin cross-linking enzyme, factor XIII (FXIII) The goal of this application is to determine the molecular mechanisms by which FXIII, fibrin(ogen), FXIII-mediated fibrin cross-linking, and RBCs contribute to venous thrombosis. Our overall hypotheses are that FXIII activity is required to retain RBCs in venous thrombi, and that blocking or reducing FXIII activity will reduce venous thrombus size. This hypothesis will be addressed in three Specific Aims: 1) Map the fibrinogen residues that mediate FXIII binding and determine the impact of FXIII activation and activity on fibrin network structure and stability, 2) Determine the contributions of blood components (FXIII, RBCs, and fibrinogen) to clot size in a whole blood milieu, and 3) Determine the effect of FXIII inhibitors on the formation and stability of venous thrombi. We will employ biochemical, genetic and pharmacologic tools in vitro and in vivo to define the FXIII-fibrinogen axis and determine the role of FXIII activity and RBC retention in venous thrombosis. Fibrin(ogen)-FXIII interactions will be examined using solution phase binding assays, microscopy, innovative fibrin analytical methods, and thromboelastography. Clot formation will be examined in reconstituted whole blood models using plasmas from healthy individuals and patients with a history of venous thrombosis. Thrombus formation and stability will be examined using venous thrombosis and thrombolysis models, immunohistochemistry, ultrasound imaging, and intravital microscopy. These studies will elucidate the biological role of FXIII activity in venous thrombosis, and define novel roles for fibrin(ogen) and RBCs in venous thrombus formation and stability. The study is highly innovative because it challenges the current paradigm that RBCs are passively trapped in static, fibrin- rich venous thrombi. The proposed research is clinically significant because it may reveal new strategies to reduce venous thrombosis in the US population.

描述（由申请人提供）：静脉血栓形成/血栓栓塞每年影响超过100万美国人。血栓负荷（大小）可预测长期结局。通过开发新药减少静脉血栓负担的努力可能对美国人群产生相当大的影响。静脉“红色”血栓的定义特征是它们的高红细胞（RBC）含量，传统上认为这是由于RBC在静态纤维蛋白凝块中的被动捕获。然而，我们新的初步数据表明，红细胞通过一种需要纤维蛋白交联酶，因子XIII（FXIII）的活性机制保留在凝块中。本申请的目的是确定FXIII，纤维蛋白（原），FXIII介导的纤维蛋白交联和红细胞有助于静脉血栓形成的分子机制。我们的总体假设是，需要FXIII活性来保留静脉血栓中的RBC，并且阻断或降低FXIII活性将减小静脉血栓大小。该假设将在三个特定目标中得到解决：1）绘制介导FXIII结合的纤维蛋白原残基并确定FXIII活化和活性对纤维蛋白网络结构和稳定性的影响，2） 确定血液成分（FXIII、RBC和纤维蛋白原）对全血环境中凝块大小的贡献，和3）确定FXIII抑制剂对静脉血栓形成和稳定性的影响。我们将在体外和体内使用生物化学、遗传学和药理学工具来定义FXIII-纤维蛋白原轴，并确定FXIII活性和RBC保留的作用。 静脉血栓形成将使用溶液相结合试验、显微镜、创新的纤维蛋白分析方法和血栓弹力图检查纤维蛋白（原）-FXIII相互作用。将使用来自健康个体和有静脉血栓形成史的患者的血浆在重建的全血模型中检查凝块形成。将使用静脉血栓形成和血栓溶解模型、免疫组织化学、超声成像和活体显微镜检查血栓形成和稳定性。这些研究将阐明FXIII活性在静脉血栓形成中的生物学作用，并确定纤维蛋白（原）和RBC在静脉血栓形成和稳定性中的新作用。这项研究是高度创新的，因为它挑战了目前的范式，即红细胞被动地被困在静态的、富含纤维蛋白的静脉血栓中。这项拟议中的研究具有临床意义，因为它可能揭示减少美国人群静脉血栓形成的新策略。

项目成果

Primed to Understand Fibrinogen in Cardiovascular Disease.

准备了解心血管疾病中的纤维蛋白原。

• DOI: 10.1161/atvbaha.115.306754
• 发表时间: 2016
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Wolberg,AlisaS

Editorial Comment: Factor XIII: One More Critical Factor for Hemostasis.

编辑点评：第十三因素：止血的又一关键因素。

• DOI: 10.1213/xaa.0000000000000154
• 发表时间: 2015
• 期刊: A & A case reports
• 作者: Wolberg,AlisaS;Levy,JerroldH
• 通讯作者: Levy,JerroldH

Fibrinogen, red blood cells, and factor XIII in venous thrombosis.

• DOI: 10.1111/jth.12918
• 发表时间: 2015-06
• 期刊: Journal of thrombosis and haemostasis : JTH
• 作者: Walton BL;Byrnes JR;Wolberg AS
• 通讯作者: Wolberg AS