Kindlin-3 Signaling in Blood Cells

血细胞中的 Kindlin-3 信号转导

• 批准号: 9193133
• 负责人: Yan-Qing Ma
• 金额: $ 41.75万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9193133
• 关键词: Affect; Animals; Applications Grants; Binding; Binding Proteins; Biochemical; Biological Models; Blood Cells; Blood Platelets; Blood coagulation; Bone Marrow; Cells; Coagulation Process; Complement; Complex; Cytoplasmic Tail; Data; Deep Vein Thrombosis; Defect; Deposition; Development; Etiology; Evaluation; Event; Functional disorder; Generations; Goals; Growth; HL60; Hemorrhage; Hereditary Disease; Immune system; Infection; Inferior vena cava structure; Infiltration; Inflammation; Inflammatory Response; Integrin beta Chains; Integrins; Kinetics; Knock-in Mouse; Lead; Leukocyte Adhesion Deficiency; Leukocytes; Ligature; Maps; Mediating; Mediator of activation protein; Modeling; Molecular; Morbidity - disease rate; Mus; Mutation; Neutrophil Activation; Neutrophil Infiltration; Patients; Play; Prevention strategy; Production; Pulmonary Embolism; Reactive Oxygen Species; Recruitment Activity; Recurrence; Reporter; Research Project Grants; Resolution; Risk Factors; Role; Signal Pathway; Signal Transduction; Site; Testing; Therapeutic; Thromboembolism; Thromboplastin; Thrombosis; Thrombus; Venous; Venous Thrombosis; base; deep vein; design; extracellular; in vivo; insight; intravital microscopy; mortality; mutant; neutrophil; new therapeutic target; novel; prevent; treatment strategy

Deep vein thrombosis (DVT) is one of the major causes of morbidity and mortality worldwide. DVT is associated with multiple risk factors and occurs when a blood clot forms in the deep veins. Significantly, recent studies have revealed that neutrophils are essentially involved in DVT formation, indicating that DVT is a complex, coordinated dysfunction of both the coagulation cascade and the innate immune system. Studies in animal DVT models have demonstrated that initial neutrophil recruitment and subsequent release of neutrophil extracellular traps (NETs) promote thrombus growth; in addition, later neutrophil infiltration facilitates thrombus resolution, suggesting that neutrophils may play dual functional roles in DVT. Kindlin-3 is a recently identified integrin β cytoplasmic tail binding protein in blood cells and required for supporting integrin-mediated leukocyte recruitment and infiltration. Patients with kindlin-3 deficiency suffer from Type-III leukocyte adhesion deficiency, a rare genetic disorder characterized by recurrent infections and severe bleeding due to integrin dysfunction in both leukocytes and platelets. Importantly, our preliminary observations have suggested that kindlin-3 also carries out a number of integrin-independent functions in neutrophils, including suppressing NET release. Therefore, we hypothesize that kindlin-3 plays multiple roles in regulating the kinetics of DVT. The overall goal of this competitive application is to examine both integrin-dependent and integrin-independent functions of kindlin-3 in neutrophils in DVT formation, and to mechanistically interpret the novel function of kindlin-3 that regulates NET release. Two Specific Aims are proposed: (1) Specific Aim 1 will determine the importance of kindlin-3/integrin interactions in neutrophils in DVT formation. We will examine the kinetics of DVT formation in a standard inferior vena cava (IVC) ligature model in mice that carry an integrin-interaction disrupting mutation in kindlin-3 or a kindlin-3 deletion specifically in neutrophils. The kinetics and extent of neutrophil recruitment, infiltration in the IVC, NET release, thrombus growth and resolution will be quantified in these mice using a combination of both intravital microscopy and histological analysis. We expect that kindlin-3 plays important roles in both initial neutrophil recruitment to inflamed IVC where venous thromboemboli develop and later neutrophil infiltration into the formed thrombi that promotes DVT resolution. (2) Specific Aim 2 will determine the molecular mechanism by which neutrophil kindlin-3 regulates NET release in DVT. The key subdomain(s) and residue(s) in kindlin-3 responsible for regulating NET release will be mapped in informative model systems. The signaling intermediates between kindlin-3 and NET release will be identified by established biochemical approaches. Further, the contribution of kindlin-3/NETs signaling pathway to DVT formation will be evaluated in mice. Taken together, these studies will redefine our understanding of the complex mechanisms by which neutrophil kindlin-3 regulates the kinetics of DVT, thus providing opportunities for designing novel and targeted therapeutic strategies for prevention and treatment of DVT.

深静脉血栓形成（DVT）是全球发病率和死亡率的主要原因之一。 DVT是 与多种危险因素相关，并在深静脉中形成血液时发生。重要的是，最近 研究表明，中性粒细胞基本上与DVT形成有关，表明DVT是一个 凝血级联和先天免疫系统的复杂，协调的功能障碍。研究 动物DVT模型已经表明，初始嗜中性粒细胞募集和随后释放中性粒细胞 细胞外陷阱（网）促进血栓生长；此外，后来的中性粒细胞浸润有助于血栓 分辨率，表明中性粒细胞可能在DVT中扮演双重功能。 Kindlin-3是最近确定的 整联蛋白β细胞质尾巴结合蛋白在血细胞中，支撑整联蛋白介导的白细胞所必需 招募和渗透。 III型白细胞粘附缺乏症患者，患有Kindlin-3缺乏症患者， 一种罕见的遗传疾病，其特征是复发性感染和因整联蛋白功能障碍引起的严重出血 白细胞和血小板。重要的是，我们的初步观察表明，Kindlin-3也 在中性粒细胞中执行许多独立的整联蛋白功能，包括抑制净释放。 因此，我们假设Kindlin-3在调节DVT的动力学中起多种作用。总体目标 这种竞争性应用程序是检查整联蛋白依赖性依赖性和整合素独立的功能 DVT形成中嗜中性粒细胞中的Kindlin-3，并机械地解释了Kindlin-3的新功能 调节净释放。提出了两个具体目的：（1）具体目标1将决定 DVT形成中嗜中性粒细胞中的Kindlin-3/整合素相互作用。我们将研究DVT形成的动力学 小鼠中标准的下腔静脉（IVC）结扎模型，该模型携带整联蛋白相互作用破坏突变 在Kindlin-3或Kindlin-3缺失中，专门在中性粒细胞中。中性粒细胞的动力学和程度， IVC，净释放，血栓生长和分辨率的浸润将在这些小鼠中使用A量化 插入显微镜和组织学分析的结合。我们希望Kindlin-3扮演很重要 在初始中性粒细胞募集到发炎的IVC中的作用 中性粒细胞浸润到促进DVT分辨率的形成的血栓中。 （2）特定目标2将决定 中性粒细胞Kindlin-3调节DVT中的净释放的分子机制。关键子域 andlin-3中负责控制净释放的住宅将在信息性模型系统中映射。 Kindlin-3和净释放之间的信号传导中间体将通过已建立的生化识别 方法。此外，将评估Kindlin-3/Nets信号通路对DVT形成的贡献 老鼠。综上所述，这些研究将重新定义我们对复杂机制的理解 中性粒细胞Kindlin-3调节DVT的动力学，从而提供了设计和针对性的机会 DVT预防和治疗的治疗策略。