Identification of compounds that reverse cellular toxicity of A beta peptide in a

描述(由申请人提供)：我们的实验室已经使用酵母来模拟与神经退行性疾病有关的人类蛋白引起的细胞缺陷。我们提出了一种高通量的化学筛选，使用一种表达人A2的酵母菌株，人类A2是一种与阿尔茨海默病相关的蛋白质。来自中试筛选的数据表明，我们的酵母菌A2检测是稳健和可重复性的。拟议的屏幕将使我们能够在多种浓度下探测更大的文库，并覆盖更广阔的化学空间。来自这一筛选的HIT化合物将针对其他神经退行性疾病的酵母模型进行反筛选，以确定选择性，并将进一步筛选对哺乳动物细胞的毒性和对哺乳动物神经元的有效性。理想的候选化合物应该在我们的初级酵母和次级神经元毒性试验中都具有活性，并且应该具有适合在转基因阿尔茨海默病小鼠模型中研究的药代动力学曲线。这些化学探针将提供一种补充方法来破译我们已经开始通过基因筛查发现的错综复杂的毒性和保护机制。 公共卫生相关性：A2肽在阿尔茨海默病患者的大脑中积聚，可能导致神经退化，但其作用机制尚不清楚。我们建议鉴定可以减轻A2的细胞毒性的化合物，并在神经元模型中验证这些化合物。这些探针将是解剖A2病理的有价值的工具，并可能成为未来临床前开发的重点。

DESCRIPTION (provided by applicant): Our laboratory has used yeast to model the cellular defects caused by the human proteins implicated in neurodegenerative diseases. We propose a high- throughput chemical screen using a yeast strain expressing human A2, a protein linked to Alzheimer's disease. Data from a pilot screen demonstrate that our yeast A2 assay is robust and reproducible. The proposed screen will enable us to probe a larger library at multiple concentrations and cover a broader chemical space. Hit compounds from this screen will be counter-screened against yeast models of other neurodegenerative diseases to determine selectivity, and will be further screened for toxicity in mammalian cells and efficacy in mammalian neurons. The ideal chemical candidates would have activity in both our primary yeast and secondary neuronal toxicity assays and would have a pharmacokinetic profile suitable for studies in transgenic mouse models of AD. These chemical probes will provide a complementary approach to deciphering the tangled mechanisms of toxicity and protection that we have begun to uncover through our genetic screens. PUBLIC HEALTH RELEVANCE: A2 peptide accumulates in the brains of patients with Alzheimer disease and may cause neurodegeneration, but its mechanism of action is not known. We propose to identify compounds that can alleviate the cellular toxicity of A2 and validate these in neuronal models. These probes will be valuable tools for dissecting A2 pathology and could potentially become the focus for future pre- clinical development.