Alpha5Beta1 Integrin inhibition as a Profound Blood-Brain Barrier Stabilizing Neuroprotective Stroke Therapy

Alpha5Beta1 整合素抑制作为深层血脑屏障稳定神经保护性中风治疗

• 批准号: 9058305
• 负责人: Gregory Jaye Bix
• 金额: $ 2万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9058305
• 关键词: Anatomy; Antineoplastic Agents; Apoptosis; Biochemical; Biological Assay; Blood - brain barrier anatomy; Blood coagulation; Blood flow; Brain; Brain Injuries; Cause of Death; Clinical Trials; Dose; Edema; Endothelial Cells; Extracellular Matrix; Genetic; Glucose; Goals; Histology; Human; Immunohistochemistry; In Vitro; Infarction; Inflammation; Injury; Integrin Inhibition; Integrins; Ischemia; Ischemic Brain Injury; Ischemic Stroke; Knock-out; Knockout Mice; Label; Link; Magnetic Resonance Imaging; Malignant Neoplasms; Mediating; Methods; Middle Cerebral Artery Occlusion; Morbidity - disease rate; Motor; Mus; Oxygen; Peptides; Permeability; Protein Fragment; Protein Isoforms; Proteins; Proteoglycan; Receptor Inhibition; Recovery of Function; Resistance; Role; Signal Transduction; Stroke; Testing; Therapeutic; Tight Junctions; Time; Treatment Efficacy; Vascular Endothelial Cell; Wild Type Mouse; analog; cell injury; cell type; cerebrovascular; deprivation; experience; in vitro Model; in vivo; inhibitor/antagonist; innovation; mouse model; novel; novel therapeutic intervention; perlecan; post stroke; prevent; public health relevance; receptor; repaired; response to injury; stroke therapy; therapeutic target; thrombolysis; treatment strategy

 DESCRIPTION (provided by applicant): Although stroke is the fourth leading cause of death in the U.S., the current treatment strategy of thrombolysis for the majority of strokes due to a blood clot (ischemic) has mixed results. Thus, there is an urgent need for new and better stroke therapies. We have previously demonstrated that the domain V protein fragment of the brain extracellular matrix component perlecan is both neuroprotective and neuroreparative after ischemic stroke by interaction with the brain endothelial cell receptor α5β1 integrin. Intriguingy, preliminary results now suggest that mice with an endothelial selective deletion of α5β1 integrin are profoundly resistant to experimental ischemic stroke; they show little to no signs of brain injury. Furthermore, inhibition of the α5β1 integrin with ATN-161 or its more potent isoform Ac-PhScN-NH2 conveys nearly identical resistance to stroke injury in wild type mice. This may occur via stabilization of the blood-brain barrier through increased function of the brain endothelial cell tight junction protein claudin-5, which in turn minimizes vasogenic edema, inflammation, and injury. Therefore, we hypothesize that endothelial cell α5β1 integrin could be a particularly effective therapeutic target for stroke. In this application, we propose the followig specific aims: 1. Determine the effect of endothelial cell selective α5β1 integrin deletion on experimental ischemic stroke, 2. Determine the potential of the α5β1 integrin as a therapeutic target in experimental ischemic stroke and 3. Determine, in mechanistic detail, the role of α5β1 integrin in modulating blood-brain barrier integrity and subsequent resistance to ischemic stroke. We will use several novel genetically modified mice (α5 integrin endothelial cell specific knockdown mice, claudin5 eGFP tagged mice) and newly characterized α5β1 integrin inhibitors in experimental stroke models and in vitro endothelial cell barrier assays. We expect to demonstrate that suppression or inhibition of α5β1 integrin in endothelial cells affords significnt blood-brain barrier-mediated resistance to experimental ischemic stroke, supporting our long-term goal of developing α5β1 integrin as a novel human stroke therapeutic target.

 描述（由申请人提供）：尽管中风是美国第四大死因，但目前对大多数因血栓（缺血性）引起的中风进行溶栓治疗的结果好坏参半。因此，迫切需要新的、更好的中风疗法。我们之前已经证明，脑细胞外基质成分基底蛋白聚糖的结构域 V 蛋白片段通过与脑内皮细胞受体 α5β1 整合素相互作用，在缺血性中风后具有神经保护和神经修复作用。有趣的是，初步结果表明，内皮细胞选择性删除 α5β1 整合素的小鼠 对实验性缺血性中风有很强的抵抗力；他们几乎没有任何脑损伤的迹象。此外，用 ATN-161 或其更有效的亚型 Ac-PhScN-NH2 抑制 α5β1 整合素，可以在野生型小鼠中产生几乎相同的对中风损伤的抵抗力。这可能是通过增加脑内皮细胞紧密连接蛋白claudin-5的功能来稳定血脑屏障来实现的，从而最大限度地减少血管源性水肿、炎症和损伤。因此，我们假设内皮细胞α5β1整合素可能是中风特别有效的治疗靶点。在本申请中，我们提出了以下具体目标：1.确定内皮细胞选择性α5β1整合素缺失对实验性缺血性中风的影响，2.确定α5β1整合素作为实验性缺血性中风治疗靶点的潜力，3.确定α5β1整合素在调节血脑屏障完整性和随后的缺血性抵抗力中的作用。 中风。我们将在实验性中风模型和体外内皮细胞屏障测定中使用几种新型转基因小鼠（α5整合素内皮细胞特异性敲除小鼠、claudin5 eGFP标记小鼠）和新表征的α5β1整合素抑制剂。我们希望证明抑制或抑制内皮细胞中的α5β1整合素可以对实验性缺血性中风提供显着的血脑屏障介导的抵抗力，支持我们将α5β1整合素开发为新型人类中风治疗靶点的长期目标。