Multiplexed Drug Discovery Assay for Neurodegeneration

描述（由申请人提供）：亨廷顿氏病是由多聚谷氨酰胺（poly-Q）束在其他不相关的蛋白质中的扩张引起的神经变性疾病家族的一个成员。含有超过40个连续谷氨酰胺残基的蛋白质形成由致病蛋白质和其它富含谷氨酰胺的细胞蛋白质组成的细胞内聚集体。虽然含poly-Q的蛋白质广泛表达，但细胞毒性主要局限于中枢神经系统。poly-Q介导的毒性机制尚未完全了解。提出的机制包括蛋白质聚集体的直接毒性和神经元细胞健康所需的富含谷氨酰胺的转录因子的耗尽。我们建议在秀丽隐杆线虫中开发用于poly-Q介导的神经退行性疾病的多重药物发现测定。该测定将同时监测两种荧光报告蛋白的神经元表达，因此将允许鉴定在专注于单个参数的测定中可能遗漏的有用的先导化合物。该检测将允许药物发现研究人员将命中定义为影响一个或两个荧光报告分子的化合物，从而增加检测的特异性。荧光报告基因将模拟：1）多聚氰胺化的致病蛋白，和2）富含谷氨酰胺的转录因子的共凝集。我们的总体I期和II期目标是通过体内筛选疾病模型来发现药物的有效方法，该方法将被授权给生物制药合作伙伴。将该测定引入药物发现实验室，有望鉴定出适合进一步临床前和临床开发的新型生物活性化合物。

DESCRIPTION (provided by applicant): Huntington's Disease is one member of a family of neurodegenerative diseases caused by expansion of polyglutamine (poly-Q) tracts in otherwise unrelated proteins. Proteins containing more than 40 consecutive glutamine residues form intracellular aggregates comprised of the disease-causing protein and other glutamine rich cellular proteins. While the poly-Q containing proteins are widely expressed, cellular toxicity is localized primarily to the central nervous system. The mechanism of poly-Q mediated toxicity is not fully understood. Proposed mechanisms include direct toxicity of protein aggregates and depletion of glutamine-rich transcription factors required for neuronal cell health. We propose to develop a multiplexed drug discovery assay in Caenorhabditis elegans for poly-Q mediated neurodegenerative disease. This assay will monitor neuronal expression of two fluorescent reporter proteins simultaneously and will thus allow identification of useful lead compounds that might be missed in assays focused on a single parameter. The assay will allow drug discovery researchers to define a hit as a compound affecting either one or both of the fluorescent reporters thereby increasing the specificity of the assay. The fluorescent reporters will model: 1) polyglutaminated disease causing proteins, and 2) co-agglutination of glutamine-rich transcription factors. Our overall Phase I and II goal is a validated method of drug discovery by in vivo screening of disease models that will be licensed to a biopharmaceutical partner. Introduction of the assay into the drug discovery lab is expected to lead to identification of novel bioactive compounds that are suitable for further preclinical and clinical development.