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

描述（由申请人提供）：我们的实验室使用酵母来模拟由涉及神经退行性疾病的人类蛋白质引起的细胞缺陷。我们提出了一个高通量的化学筛选，使用酵母菌株表达人类A2，一种与阿尔茨海默氏病有关的蛋白质。来自中试筛选的数据表明，我们的酵母A2测定是稳健的和可重现的。拟议的屏幕将使我们能够探测更大的图书馆在多个浓度和覆盖更广泛的化学空间。来自该筛选的命中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，并将进一步筛选哺乳动物细胞中的毒性和哺乳动物神经元中的功效。理想的化学候选物将在我们的初级酵母和次级神经元毒性测定中具有活性，并且将具有适合于在AD转基因小鼠模型中研究的药代动力学特征。这些化学探针将提供一种互补的方法来破译我们已经开始通过基因筛选发现的毒性和保护的复杂机制。 公共卫生相关性：A2肽在阿尔茨海默病患者的大脑中积累，可能导致神经退行性变，但其作用机制尚不清楚。我们建议确定化合物，可以减轻A2的细胞毒性，并验证这些神经元模型。这些探头将成为解剖A2病理学的宝贵工具，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.