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: 8630620
• 负责人: Anil Kumar Chauhan
• 金额: $ 37.75万
• 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/8630620
• 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; public health relevance; restoration; tegrin; toll-like receptor 4; translational approach; 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; public health relevance; 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（由适用提供）：鉴定新型内源配体，这些配体在病理环境中促进血栓形成和炎症过程，例如糖尿病，高胆固醇血症和血管高血压，可能会导致新的疗法，从而改善中风风险高的患者。含有替代性的额外结构域A（EDA+-FN）的纤维蛋白（FN）的变体在健康人类和小鼠的动脉或循环中不存在，在糖尿病动脉的森林中特异性表达，并且在糖尿病上的疾病中，在疾病中的循环中升高，可能会升高，这可能是糖尿病的肿瘤，并且有可能催眠效率。最近，我们发现EDA+-FN促进血栓形成和炎症过程。 EDA+-FN有助于血栓形成和炎症的基本机制尚不清楚。已知EDA+-FN会激活Toll-Like-Thepepor 4（TLR4）信号通路。此外，EDA是整联蛋白€9¿1的配体，该配体在炎症细胞（例如中性粒细胞和巨噬细胞）上表达。高胆固醇血症是人类急性中风的主要危险因素之一。因此，在高胆固醇血症的载脂蛋白E缺乏（APOE - / - ，动脉粥样硬化容易发生）小鼠中，我们建议测试EDA+-FN的中心假设，即EDA+-FN有助于缺血/再生灌注（I/R）在病理环境中及其在病理环境中及其有助于 通过平行€9和TLR4介导的途径来增强血栓形成和炎症过程。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，我们将确定整联蛋白»9€1在EDA+-FN介导的血栓形成和炎症I/R脑损伤中的作用。在AIM 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。此外，我们将确定优先条件的等离子体中EDA+-FN的来源。作为一种翻译方法，我们将检验以下假设：用特定的单克隆抗体阻断EDA+-FN将在高胆固醇血症的背景下减少中风损伤。为了实现我们的特定实验目标，我们开发了新型的转基因小鼠菌株和试剂，我们将与该领域的其他研究人员共享。提出的研究的贡献在临床上具有很高的临床意义，因为它决定了机械见解 EDA+-FN在动脉粥样硬化的疾病背景下促进血栓形成和炎症性脑损伤。该提案具有未来的翻译潜力，因为它可能会产生重大影响 关于诊断和治疗常见的血栓炎性疾病，包括急性中风。