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。血栓负担（大小）预测了长期结果。通过开发新药来减轻静脉血栓负担的努力可能会对美国人口产生重大影响。静脉“红色”血栓的定义特征是它们的高红细胞（RBC）含量，传统上被认为是由于静态纤维蛋白血块中RBC的被动捕获而引起的。 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.我们的总体假设是，将FXIII活性保留在静脉血栓中，而阻塞或减少FXIII活性将减少静脉血栓大小。该假设将以三个具体目的解决：1）绘制介导FXIII结合的纤维蛋白原残基，并确定FXIII激活和活动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）对整个血液中的凝块大小的贡献，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化，遗传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 在静脉血栓形成中。将使用溶液相结合测定，显微镜，创新的纤维蛋白分析方法和血栓纤维射击检查纤维蛋白（OGEN）-FXIII相互作用。使用具有静脉血栓形成史的健康个体和患者的血浆中，将在重构的全血模型中检查凝块的形成。血栓形成和稳定性将使用静脉血栓形成和溶栓模型，免疫组织化学，超声成像和浸润显微镜检查。这些研究将阐明FXIII活性在静脉血栓形成中的生物学作用，并定义纤维蛋白（OGEN）和RBC在静脉血栓形成和稳定性中的新作用。这项研究具有高度创新性，因为它挑战了当前RBC被动地陷入静态，纤维蛋白丰富的静脉血栓的范例。拟议的研究具有临床意义，因为它可能揭示了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

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

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