Astrocytic LRP-1 Modulates Blood-Brain Barrier Function

星形胶质细胞 LRP-1 调节血脑屏障功能

• 批准号: 9898289
• 负责人: Manuel Salvador Yepes
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9898289
• 关键词: Acute; Adhesions; Admission activity; Alteplase; Animal Model; Astrocytes; Basement membrane; Binding; Biological Assay; Blood - brain barrier anatomy; Brain; Caring; Cause of Death; Cell Adhesion; Cell Culture Techniques; Cell surface; Cerebral Edema; Cerebral Ischemia; Cerebral hemisphere hemorrhage; Cleaved cell; Co-Immunoprecipitations; Collagen Type IV; Complication; Confocal Microscopy; Cytoplasmic Tail; Data; Development; Edema; Endothelial Cells; Engineering; Extracellular Domain; Extracellular Matrix; Family; Fibronectins; Foot Process; Gelatinase B; Goals; Guidelines; Hemorrhage; Hospitals; Inflammatory; Integrin Binding; Integrins; Intravenous; Ischemic Stroke; LDL-Receptor Related Protein 1; Laminin; Ligands; Light; Lipoprotein Binding; Lipoprotein Receptor; Low Density Lipoprotein Receptor; Membrane; Metalloproteases; Methods; Morbidity - disease rate; Patients; Pharmacology; Physiological; Plasminogen Activator; Proteins; Receptor Signaling; Recombinants; Research; Serine Protease; Signal Pathway; Signal Transduction; Stroke; Structure; Testing; Therapeutic; Tight Junctions; Time; Tissues; adhesion receptor; blood-brain barrier function; blood-brain barrier permeabilization; cytokine; improved; in vitro Model; inhibitor/antagonist; laminin-1; member; mortality; nanocarrier; preservation; prevent; proteinase In; receptor; recruit; response; stroke patient

Objective: The main goal of this application is to study the mechanism whereby the low density lipoprotein receptor-related protein 1 (LRP1) modulates the permeability of the BBB under physiological conditions and after an ischemic stroke. Methods In Aim I we will test the hypothesis that under non-ischemic conditions astrocytic LRP1 preserves the integrity of the basement membrane-astrocyte (BM-A) interface by promoting the recruitment of integrins to the membrane of perivascular astrocytic end- feet processes and their binding to its ligands in the basement membrane (BM). In Aim II we propose that this function is lost under ischemic conditions when the cleavage of the extracellular domain of LRP1 from perivascular astrocytes activates a cell-signaling pathway that increases the permeability of the blood-brain barrier (BBB). In Aim III we will study in an animal model of ischemic stroke whether inhibition of LRP1 cleavage from perivascular astrocytes with the receptor-associated protein (RAP) prevents the harmful effects of endogenous tPA and/or rtPA on the permeability of the BBB. Research Plan In the first Aim of this application we will use astrocytic cell cultures and a cell adhesion assay. In the second Aim we will use astrocytic cultures, an in vitro model of the blood- brain barrier, real-time PCR, confocal microscopy and co-immunoprecipitation assays. In the third Aim we will use a nanocarrier that we engineered to selectively deliver its content to the BBB in the ischemic tissue and an animal model of cerebral ischemia.

客观的： 该应用的主要目标是研究低密度的机制 脂蛋白受体相关蛋白 1 (LRP1) 调节 BBB 的通透性 在生理条件下和缺血性中风后。 方法 在目标 I 中，我们将检验以下假设：在非缺血条件下星形胶质细胞 LRP1 通过以下方式保持基底膜-星形胶质细胞 (BM-A) 界面的完整性 促进整合素募集至血管周围星形细胞终末膜 脚突及其与基底膜 (BM) 中配体的结合。瞄准II 我们认为，当缺血条件下，当 血管周围星形胶质细胞 LRP1 的胞外结构域激活细胞信号传导 增加血脑屏障（BBB）通透性的途径。在目标 III 中，我们 将在缺血性中风动物模型中研究是否抑制 LRP1 裂解 来自血管周围星形胶质细胞的受体相关蛋白（RAP）可防止 内源性 tPA 和/或 rtPA 对 BBB 通透性的有害影响。 研究计划 在此应用的第一个目标中，我们将使用星形胶质细胞培养物和细胞粘附 化验。在第二个目标中，我们将使用星形细胞培养物，这是一种血液的体外模型 脑屏障、实时 PCR、共聚焦显微镜和免疫共沉淀分析。在 第三个目标，我们将使用我们设计的纳米载体来选择性地传递其 缺血组织中血脑屏障的含量及脑缺血动物模型。