Multiplexed Drug Discovery Assay for Neurodegeneration

描述(由申请人提供)：亨廷顿病是一种神经退行性疾病家族的成员，由多聚谷氨酰胺(Poly-Q)束在其他无关的蛋白质中扩张引起。含有40多个连续谷氨酰胺残基的蛋白质形成细胞内聚集体，由致病蛋白和其他富含谷氨酰胺的细胞蛋白组成。虽然含有多聚Q的蛋白质被广泛表达，但细胞毒性主要局限于中枢神经系统。多聚Q介导的毒性机制尚不完全清楚。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。我们建议在秀丽线虫中建立一种针对多Q介导的神经退行性疾病的多重药物发现方法。这种分析将同时监测两种荧光报告蛋白的神经元表达，从而能够识别在专注于单一参数的分析中可能遗漏的有用的先导化合物。该分析将允许药物发现研究人员将HIT定义为影响其中一种或两种荧光报告的化合物，从而增加了分析的特异性。荧光记者将模拟：1)多谷氨酰胺引起疾病的蛋白质，以及2)富含谷氨酰胺的转录因子的共凝集。我们第一阶段和第二阶段的总体目标是通过体内筛选疾病模型来验证药物发现的方法，这些疾病模型将被授权给生物制药合作伙伴。将该分析引入药物发现实验室有望鉴定出适合于进一步临床前和临床开发的新的生物活性化合物。

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.