Vascular Pannexin 1 in Ischemic Stroke

缺血性中风中的血管 Pannexin 1

基本信息

批准号：
9761559
负责人：
Miranda E Good
金额：
$ 10.32万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-15 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9761559
关键词：
Academia Acute Adhesions Area Arteries Blood Vessels Blood flow Bone Marrow Transplantation Cell Adhesion Cells Cerebral Ischemia Cerebrovascular Circulation Cerebrovascular system Cerebrum Cessation of life Data Development Disease Emigrations Endothelial Cells Endothelium Etiology Experimental Designs Flow Cytometry Fostering Foundations Future Genetic Goals Image Infarction Inflammation Inflammatory Injury Integrins Intervention Ischemia Ischemic Stroke Knockout Mice Knowledge Lead Leukocytes Mentors Mentorship Mesenteric Arteries Morbidity - disease rate Mus Neurologic Deficit Nucleotides Obstruction Peripheral Pharmaceutical Preparations Pharmacology Pharmacotherapy Phase Play Preparation Process Protein Isoforms Proteins Purinoceptor Receptor Signaling Recovery Regulation Reperfusion Injury Reperfusion Therapy Research Research Personnel Role Severities Signal Transduction Stroke TNF gene Techniques Testing Therapeutic Intervention Thrombolytic Therapy Time Tissues Training Transgenic Mice Venous Work autocrine brain endothelial cell career development cerebral artery cerebral vein cost experience experimental study improved in vivo in vivo imaging insight intravital microscopy mechanotransduction mortality new therapeutic target novel overexpression post stroke programs purinoceptor P2Y4 receptor response skills stroke outcome stroke patient targeted treatment therapeutic target venule

项目摘要

|| PROJECT SUMMARY/ABSTRACT Ischemic stroke is a significant disease that can lead to neurological deficits or death; however, only one pharmacological intervention drug is available and it predominately aims to eliminate the physical occlusion within the blood vessel. To explore potential therapeutic targets to improve recovery following an ischemic stroke, we began examining the role of Panx1 channels, which activate downstream purinergic receptors, within the cerebral vasculature. I have recently shown that deletion of endothelial Panx1 profoundly improves post-cerebral ischemia/reperfusion infarct volume. However, endothelial Panx1 could significant contribute to both arterial and venous endothelial functions. In this application, we demonstrate that arterial myogenic tone development (arterial), and inflammation of leukocytes following an ischemic stroke (venous), are significantly decreased in mice lacking endothelial Panx1. This application tests the hypothesis that endothelial Panx1 contributes to the development of ischemic stroke injury through both regulation of cerebral arterial myogenic tone and regulation of leukocyte adhesion and emigration in cerebral venous endothelial cells. Our preliminary data suggest that Panx1 activation during myogenic tone development is regulated through interaction with b3 integrins. Whereas, in cerebral veins, I find leukocyte adhesion in cerebral venules following an ischemic stroke is regulated through Panx1 channel function. These studies will provide insight into the differential roles and regulation of Panx1 within arterial and venous cerebral endothelial cells, their contribution to the etiology of ischemic stroke injury, and lay the foundation for future identification of therapeutic interventions for ischemic stroke patients. During the mentored phase of this application, I will develop novel transgenic mice to evaluate the overexpression of endothelial Panx1 and obtain necessary skills to evaluate the functional consequence of Panx1-b3 interaction and image cerebral inflammation following an ischemic stroke using in vivo intravital microscopy techniques. I will continue my professional and scientific development in preparation for the independent phase with continuous guidance from my mentoring committee. During the independent phase, I will develop a novel transgenic mouse that will delete or overexpress Panx1 specifically within arterial endothelial cells and determine the role of arterial verse venous endothelial Panx1 during an ischemic stroke. In addition, I will establish the downstream purinergic signaling involved in Panx1-dependent leukocyte adhesion, by in vivo imaging of cerebral venules, and emigration, using flow cytometry, and its impact of determining the severity of an ischemic stroke. This project will foster my continued scientific and professional training and facilitate my establishing an independent research program in academia.

||项目概要/摘要缺血性中风是一种严重的疾病，可导致神经功能缺损或死亡；然而，只有一个药物干预药物是可用的，其主要目的是消除物理闭塞血管内。探索潜在的治疗靶点以改善缺血后的恢复中风后，我们开始研究 Panx1 通道的作用，该通道激活下游嘌呤能受体，脑血管系统内。我最近发现内皮 Panx1 的缺失可以显着改善脑缺血/再灌注后梗塞体积。然而，内皮 Panx1 可能对动脉和静脉内皮功能。在此应用中，我们证明了动脉肌源性张力发育（动脉）和缺血性中风（静脉）后白细胞炎症显着在缺乏内皮 Panx1 的小鼠中，该值下降。该应用测试了内皮 Panx1 的假设通过调节脑动脉肌源性促进缺血性中风损伤的发展脑静脉内皮细胞中白细胞粘附和迁移的张力和调节。我们的初步数据表明，肌原性张力发育过程中 Panx1 的激活是通过与 b3 相互作用来调节的整合素。然而，在脑静脉中，我发现缺血后脑小静脉中的白细胞粘附行程通过 Panx1 通道功能进行调节。这些研究将深入了解不同的角色 Panx1 在动脉和静脉脑内皮细胞内的调控及其对病因学的贡献缺血性中风损伤，为未来确定缺血性中风治疗干预措施奠定基础中风患者。在本申请的指导阶段，我将开发新型转基因小鼠来评估内皮 Panx1 的过度表达并获得评估功能后果的必要技能 Panx1-b3 相互作用以及利用体内活体成像对缺血性中风后的脑炎症进行成像显微镜技术。我将继续我的专业和科学发展，为在我的指导委员会的持续指导下的独立阶段。在独立阶段，我将开发一种新型转基因小鼠，该小鼠将在动脉内特异性删除或过度表达 Panx1 内皮细胞并确定动脉与静脉内皮 Panx1 在缺血性中风期间的作用。此外，我将建立 Panx1 依赖性白细胞中涉及的下游嘌呤能信号传导通过脑小静脉的体内成像观察粘附性，并使用流式细胞术观察迁移性及其影响确定缺血性中风的严重程度。这个项目将培养我继续科学和专业培训并促进我在学术界建立独立的研究计划。