Discovering Novel Drug Targets for CNS Edema by In Vitro Genetics

摘要： 脑肿胀是创伤性脑损伤(TBI)、缺血性中风等严重并发症的一种 条件。仅在美国，每年就有140万人和70万人遭受脑外伤和中风的折磨。他们是一种昂贵的健康 护理负担和毁灭性的社会负担。目前脑肿胀的治疗方法是有限的，通常 无效，突出了对更好的治疗方法的戏剧性的未得到满足的需求。更好地理解 迫切需要分子途径和细胞机制来识别新的药物靶点和预测 可以对患者进行分层以做出临床治疗决策的生物标志物。该项目的目标是确定 星形胶质细胞细胞毒性水肿反应的新药物靶点和生物标志物。我们将用一部小说和 我们开发的创新技术，采用不偏不倚的方法在功能识别 脑细胞肿胀的原因介体。我们的方法使用了大量遗传多样性的星形胶质细胞 以确定细胞毒性水肿的机械基础上的基因和途径。在第一阶段，我们 建立了一种高通量星形胶质细胞肿胀动力学分析方法，该方法具有较强的稳定性和可扩展性，可用于筛选 引起或阻止肿胀的化合物。在第二阶段，我们将筛选300个遗传多样性的星形胶质细胞 然后绘制和识别对细胞肿胀的诱导剂和阻断剂的中介反应的基因。 候选目标基因的验证将在人类和小鼠星形胶质细胞中进行。人类基因 其改变了人类星形胶质细胞反应，也是新药的治疗可及的靶基因 发现号是这个项目的最终目标和产品。

Abstract: Brain swelling is a serious complication of traumatic brain injury (TBI), ischemic stroke and other serious conditions. TBI and stroke afflict 1.4M and 700K persons per year in the US alone. They are a costly health care burden and a devastating social burden. Current treatments for brain swelling are limited and generally ineffective, highlighting the dramatic unmet need for better therapeutics. A better understanding of the molecular pathways and cellular mechanisms is sorely needed to identify new drug targets and predictive biomarkers that can stratify patients for clinical treatment decisions. The goal of this project is to identify new drug targets and biomarkers of response for cytotoxic edema of astrocytes. We will use a novel and innovative technology that we have developed that takes an unbiased approach to functionally identifying the causal mediators of brain cell swelling. Our approach uses a large panel of genetically diverse astrocyte lines to identify the genes and pathways that mechanistically underlie cytotoxic edema. In Phase I, we developed a high throughput kinetic assay for astrocyte swelling that is robust and scalable for screening compounds that induce or block swelling. In Phase II we will screen 300 genetically diverse astrocyte cell lines and then map and identify the genes that mediate response to inducers and blockers of cell swelling. Validation of candidate target genes will be conducted in both human and mouse astrocytes. Human genes that modify human astrocyte response that are also therapeutically accessible target genes for novel drug discovery are the ultimate aims and products of this project.