Fibronectin alternative splicing in thrombosis and inflammation

纤连蛋白选择性剪接在血栓形成和炎症中的作用

• 批准号: 8787784
• 负责人: Anil Kumar Chauhan
• 金额: $ 37.18万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8787784
• 关键词: Accounting; Acute; Alteplase; Alternative Splicing; Apolipoprotein E; Arteries; Atherosclerosis; Binding; Blood Circulation; Blood Platelets; Blood Vessels; Blood flow; Bone Marrow; Brain Injuries; Carotid Artery Thrombosis; Cell Adhesion Molecules; Cells; Cerebrum; Cessation of life; Chimera organism; Clinical; Coagulation Process; Diabetes Mellitus; Diagnosis; Disease; Endothelial Cells; Endothelium; Exons; Experimental Animal Model; Fab Immunoglobulins; Fibrinolysis; Fibronectins; Functional disorder; Future; Genes; Goals; Health; Health Care Costs; Human; Hypertension; Infarction; Inflammation; Inflammatory; Injury; Integrins; Ischemia; Ischemic Penumbra; Ischemic Stroke; Knockout Mice; Lead; Life; Ligands; Liver; Measures; Mediating; Modeling; Monoclonal Antibodies; Mouse Strains; Mus; Neurological outcome; Neurons; Outcome; Pathway interactions; Patients; Pilot Projects; Plasma; Platelet aggregation; Process; RNA Splicing; Reagent; Recruitment Activity; Reperfusion Injury; Reperfusion Therapy; Research Personnel; Risk Factors; Role; Signal Pathway; Signal Transduction; Source; Stroke; Testing; Thrombosis; Thrombus; Time; Translating; United States; Variant; Vent; Work; acute stroke; clinically significant; common treatment; defined contribution; effective therapy; endothelial dysfunction; high risk; hypercholesterolemia; improved; in vivo; inflammatory marker; insight; intravital microscopy; knowledge of results; macrophage; mouse toll-like receptor 4; mutant; neutrophil; novel; novel therapeutic intervention; overexpression; pre-clinical; restoration; tegrin; toll-like receptor 4; translational approach

DESCRIPTION (provided by applicant): Identifying novel endogenous ligands that promote thrombosis and inflammatory processes in pathological settings, such as, diabetes, hypercholesterolemia and vascular hypertension may lead to new therapies that improve outcomes in patients at high risk for stroke. A variant of fibronectin (FN) containing the alternatively-spliced extra domain A (EDA+-FN), which is absent in the arteries or circulation of healthy humans and mice, is specifically expressed in the endothelium of atherosclerotic arteries and elevated in circulation during pathological settings, such as diabetes, atherosclerosis and vascular hypertension, most likely due to endothelial dysfunction. Recently, we have discovered that EDA+-FN promotes thrombosis and inflammatory processes. The underlying mechanisms by which EDA+-FN contributes to thrombosis and inflammation are not well understood. EDA+-FN is known to activate the toll-like- receptor 4 (TLR4) signaling pathway. Additionally, EDA is a ligand for integrin ¿9¿1, which is expressed on inflammatory cells, such as, neutrophils and macrophages. Hypercholesterolemia is one of the major risk factors for acute stroke in humans. Therefore, in hypercholesterolemic apolipoprotein E-deficient (ApoE-/-, atherosclerosis prone) mice, we propose to test the central hypothesis that EDA+-FN contributes to ischemia/reperfusion (I/R) brain injury in pathological setting, and that it does so by enhancing thrombosis and inflammatory processes via parallel ¿9¿1and TLR4-mediated pathways. In Aim1, we will define the role of TLR4 in EDA+-FN-mediated thrombosis and inflammatory I/R brain injury. In Aim 2, we will determine the role of integrin ¿9¿1 in EDA+-FN -mediated thrombosis and inflammatory I/R brain injury. In Aim 3, we will define the role of plasma versus endothelial cell EDA+-FN in I/R brain injury. Furthermore, we will determine the source of EDA+-FN in the plasma of the aforementioned pathological conditions. As a translational approach, we will test the hypothesis that blocking EDA+-FN with specific monoclonal antibodies will reduce stroke injury in the context of hypercholesterolemia. To achieve our specific experimental goals, we have developed novel genetically modified mice strains and reagents that we will share with other researchers in the field. The contribution of th proposed studies is highly clinically significant as it determine the mechanistic insights by which EDA+-FN promotes thrombosis and inflammatory brain injury in disease context of atherosclerosis. The proposal has future translational potential, as it may have significant impact on the diagnosis and treatment of common thrombo-inflammatory diseases including acute stroke.

描述（由申请人提供）：鉴定在糖尿病、高胆固醇血症和血管性高血压等病理环境中促进血栓形成和炎症过程的新型内源性配体可能会带来改善中风高风险患者预后的新疗法。 (FN)含有选择性剪接的额外结构域A(EDA+-FN)，健康人和小鼠的动脉或循环中不存在该结构域，特异表达于动脉粥样硬化的内皮细胞和在糖尿病、动脉粥样硬化和血管性高血压等病理情况下血液循环升高，很可能是由于内皮功能障碍。最近，我们发现 EDA+-FN 促进血栓形成和炎症过程。 FN 会导致血栓形成，而 EDA+-FN 会激活 Toll 样受体 4 (TLR4) 信号通路，目前尚不清楚。 EDA 是整合素的配体 ¿ 9¿1，在炎症细胞（例如中性粒细胞和巨噬细胞）上表达，是人类急性中风的主要危险因素之一，因此，在高胆固醇血症载脂蛋白 E 缺陷（ApoE-/-，易发生动脉粥样硬化）小鼠中。 ，我们建议检验 EDA+-FN 导致病理环境下缺血/再灌注 (I/R) 脑损伤的中心假设，以及那它 通过并行 ¿ 增强血栓形成和炎症过程来实现这一点9¿1 和 TLR4 介导的途径 在 Aim1 中，我们将定义 TLR4 在 EDA+-FN 介导的血栓形成和炎症 I/R 脑损伤中的作用。在 Aim 2 中，我们将确定整合素 ¿ 9¿1 在 EDA+-FN 介导的血栓形成和炎症性 I/R 脑损伤中的作用 在目标 3 中，我们将定义血浆与内皮细胞 EDA+-FN 在 I/R 脑损伤中的作用。作为一种转化方法，我们将测试用特定单克隆抗体阻断 EDA+-FN 将减少中风损伤的假设。为了实现我们的具体实验目标，我们开发了新型转基因小鼠品系和试剂，我们将与该领域的其他研究人员分享这些研究的贡献具有高度的临床意义，因为它确定了其机制的见解。 EDA+-FN 在动脉粥样硬化疾病中促进血栓形成和炎症性脑损伤。该提案具有未来的转化潜力，因为它可能会产生重大影响。 擅长急性脑卒中等常见血栓炎症性疾病的诊断和治疗。