Targeting the Hippo signaling at the Blood-brain barrier for therapy of ischemic stroke

摘要 虽然已经做了相当大的努力，集中在神经元/胶质细胞损伤，以寻找新的靶点， 中风治疗，没有取得成功。中风的治疗仍然局限于预防性治疗， 急性血栓溶解和颅内压症状控制。大脑层面的病理变化 内皮细胞在脑疾病的进展中起重要作用。然而，调节的方法 用于治疗BBB功能仍然非常有限。最近，研究表明，与YES相关的 蛋白（雅普）及其辅因子TAZ（具有PDZ结合基序的转录共激活因子），转录 作为Hippo信号通路的辅活化剂，促进内皮细胞增殖、迁移和血管生成。 雅普/TAZ还对紧密连接的形成和完整性起调节作用，这表明其可能是一种重要的调节蛋白。 在BBB紧密连接中的作用。我们的中心假设是雅普/TAZ促进脑血管生成， 在维持血脑屏障完整性方面起重要作用，雅普/TAZ-外泌体具有治疗作用 通过促进BBB的修复和BBB完整性的恢复来对抗脑缺血/再灌注损伤。 本研究旨在阐明雅普/TAZ在调节血脑屏障功能中的作用， 研究雅普/TAZ对脑缺血再灌注损伤的脑EC和BBB的保护作用， 雅普/TAZ-外泌体治疗缺血性卒中的新方法我们将使用缺血性中风模型， 体外和体内来验证我们的假设 我们预期雅普/TAZ能促进脑内ECs增殖、迁移和血管生成，并能过度抑制脑内ECs的增殖、迁移和血管生成。 雅普/TAZ和雅普/TAZ-Exosomes的表达促进缺血性损伤后的BBB修复，这导致缺血性损伤后的BBB修复。 缺血性卒中结局改善，表现为行为结局改善，炎症反应减少 以及脑中的BBB渗漏等。这项研究具有高度的意义和创新性，因为它将 进一步了解雅普/TAZ在调节血脑屏障功能和脑血管病病理生理学中的作用。 缺血性卒中下的BBB。该项目将探索雅普/TAZ作为一种新的治疗靶点， 缺血性中风治疗，外泌体作为一种新的递送载体，以及脑EC作为一种新的脑血管病治疗位点。 疾病治疗和脑部药物输送。此外，所提出的治疗方法可以广泛地 用于治疗影响血脑屏障和中枢神经系统的疾病。因此，总体影响 该项目在推进针对脑部疾病的脑内皮药物发现方面具有重要意义 疗法

Abstract Although considerable efforts have been made focusing on the neuronal/glial injury to find new targets for stroke therapy, no success has been achieved. Therapy of stroke is still limited to preventive treatments and to acute thrombolysis and symptomatic control of intracranial pressure. Pathological changes at the level of brain endothelium plays an important role in progression of brain diseases. However, the approaches to modulate the BBB function for therapy are still very limited. Recently, studies have shown that the YES-associated protein (YAP) and its paralog TAZ (transcriptional co-activator with PDZ-binding motif), the transcriptional coactivators of the Hippo signaling pathway, promote endothelial cell proliferation, migration and angiogenesis. YAP/TAZ also plays a role in regulating formation and integrity of tight-junction, which indicates its possible role in tight-junction of the BBB. Our central hypothesis is that YAP/TAZ promotes brain angiogenesis and plays an important role in maintaining the BBB integrity, and YAP/TAZ-exosome has a therapeutic effect against brain ischemia/reperfusion injury by promoting repair of the BBB and restoration of the BBB integrity. The present proposal aims to elucidate the role of YAP/TAZ in regulating the BBB function, to evaluate protective effect of YAP/TAZ on brain ECs and the BBB against ischemia/reperfusion injury, and to develop a novel approach, YAP/TAZ-exosome, for treatment of ischemic stroke. We will use the ischemic stroke model in vitro and in vivo to test our hypothesis We expect that YAP/TAZ promotes brain ECs proliferation, migration and angiogenesis, and over- expression of YAP/TAZ and YAP/TAZ-Exosomes promote the BBB repair after ischemic injury, which leads to improvement of ischemic stroke outcomes shown as improved behavior outcome, less inflammatory responses and the BBB leakage in the brain etc. The proposed study is highly significant and innovative because it will advance our understanding of the role of YAP/TAZ in regulating the BBB function and pathophysiology of the BBB under ischemic stroke. The proposed project will explore YAP/TAZ as a novel therapeutic target for ischemic stroke therapy, exosome as a novel delivery carrier, and the brain EC as a novel site for brain disease therapy and brain drug delivery. Furthermore, the proposed therapeutic approach could be broadly applied for therapy of diseases affecting the BBB and central nervous system. Therefore, the overall impact of the project is highly significant in advancing drug discovery that targets the brain endothelium for brain disease therapy.