Role of oxidative stress for neuro-vascular function

活性氧簇(ROS)在脑缺血再灌流中起着有害的作用，而脑缺血是目前治疗缺血性中风的标准疗法。然而，这种“再灌注损伤”的确切血管和细胞机制仍不清楚，这主要是因为缺乏体内ROS的测量方法，以及缺乏能够研究ROS在时间和空间上的受控诱导的动物模型。目前的项目将使用新的化学遗传学技术在体外和体内以细胞和细胞器特异性的方式测量和诱导ROS。这些化学发生工具将与体内双光子显微镜、单细胞转录本和基于细胞的药物筛选平台结合使用，以1)测量小鼠中风模型神经血管单位(NVU)细胞中的ROS，以确定脑缺血再灌注期间ROS产生的细胞来源，2)诱导健康动物NVU中ROS的产生，以确定ROS对神经血管功能的特定作用，3)确定由细胞特异性ROS产生诱导的基因，以及4)确定防止或抑制ROS产生的药物。本项目的结果将确定脑缺血后ROS产生的时间和细胞特征，破译潜在的基因表达，并识别有效的抗氧化化合物，从而确定新的分子和细胞靶点和潜在的治疗中风患者的药物。

Reactive oxygen species (ROS) play a detrimental role upon reperfusion from cerebral ischemia, the current standard therapy for ischemic stroke. However, the exact vascular and cellular mechanisms of this "reperfusion injury" remain unknown mainly due to the lack of methodology to measure ROS in vivo and the lack of animal models that allow to investigate the controlled temporal and spatial induction of ROS. The current project will use novel chemogenetic technology to measure and induce ROS in a cell and organelle-specific manner in vitro and in vivo. These chemogenetic tools will be used in combination with in vivo 2-photon microscopy, single-cell transcriptomics, and a cell-based drug screening platform to 1) measure ROS in a mouse stroke model in cells of the neurovascular unit (NVU) to identify the cellular source of ROS production during reperfusion from cerebral ischemia, 2) to induce ROS production in the NVU of healthy animals to identify the specific role of ROS for neurovascular function, 3) to identify genes induced by cell-specific ROS production, and 4) to identify drugs which prevent or suppress ROS production. The results of the current project will identify the temporal and cellular profile of ROS production after cerebral ischemia, decipher the underlying gene expression, and identify effective antioxidative compounds thereby defining novel molecular and cellular targets and potential drugs for the treatment of stroke patients.