Discovering Novel Drug Targets for CNS Edema by In Vitro Genetics

摘要： 脑肿胀是创伤性脑损伤（TBI）、缺血性脑卒中等严重脑损伤的严重并发症 条件仅在美国，每年就有140万人和70万人遭受TBI和中风的折磨。这是一种昂贵的健康 社会负担和沉重的社会负担。目前脑肿胀的治疗方法有限， 无效，突出了对更好的治疗方法的巨大未满足的需求。更好地理解 分子途径和细胞机制是迫切需要的，以确定新的药物靶点和预测 生物标志物可以对患者进行分层，用于临床治疗决策。本项目的目标是确定 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。我们将使用一本小说， 我们开发的创新技术， 脑细胞肿胀的原因我们的方法使用了大量基因多样的星形胶质细胞 线，以确定基因和途径，机械基础细胞毒性水肿。在第一阶段，我们 开发了一种用于星形胶质细胞肿胀的高通量动力学测定，该测定对于筛选是稳健的且可扩展的 引起或阻止肿胀的化合物。在第二阶段，我们将筛选300个基因多样的星形胶质细胞 然后绘制并鉴定介导对细胞肿胀诱导剂和阻断剂反应的基因。 候选靶基因的验证将在人和小鼠星形胶质细胞中进行。人类基因 其也是治疗上可获得新药靶基因 发现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.