Stem Cell Panels for Genetic Analysis of Variable Toxicant Effects on Neuronal Ce

我们建议开发一种高通量的体外检测方法来鉴定遗传因素 影响毒物对神经元分化的影响。这个项目使用了一种新的基因 预测生物学开发的映射资源，多样性远交（DO）小鼠 ES细胞板。使用强大的ES细胞工程方法，我们将整合神经元 将报告基因分化为DO ES系。神经干细胞（NSC）系将被衍生 从修改的DO ES线的面板，和NSC面板将用于量化 毒物暴露后的神经元分化。这些数据将用于绘制基因座， 鉴定介导对神经毒物的可变反应的基因。项目设计 优先考虑快速、灵敏和高度可扩展的检测技术， 载体设计的方法，以促进构建多个测定组来跟踪 对特定类型或类别的神经元和神经元细胞的毒性作用。这 技术将能够在HTS规模上对大量化合物进行初步筛选， 并且将用于优先选择化合物用于体外和体内的后续研究 使用小鼠和人类细胞以及活体动物模型。的基因和 使用这项技术确定的毒理学途径将最终改善人类 通过提供对人类发育的更详细的机械理解， 神经毒性，并将有助于遗传知情的方法来估计风险 与接触神经毒物有关 !

We propose to develop a high throughput in vitro assay to identify genetic factors affecting toxicant impact on neuronal differentiation. This project uses a novel genetic mapping resource developed at Predictive Biology, the Diversity Outbred (DO) Mouse ES Cell Panel. Using robust ES cell engineering methods, we will incorporate neuronal differentiation reporters into DO ES lines. Neuronal Stem Cell (NSC) lines will be derived from the panel of modified DO ES lines, and the NSC panel will be used to quantify neuronal differentiation after toxicant exposure. These data will be used to map loci and identify genes that mediate variable response to neurotoxicants. The project design prioritizes rapid, sensitive and highly scalable assay technology, and features a cassette approach to vector design to facilitate construction of multiple assay panels to track toxicant effects on specific types or classes of neurons and neuronal cells. This technology will enable primary screening of large numbers of compounds at HTS scale, and will be useful for prioritizing compounds for follow up studies in vitro and in vivo using both mouse and human cells as well as live animal models. The genes and toxicological pathways identified using this technology will ultimately improve human health by providing a more detailed mechanistic understanding of human developmental neurotoxicity, and will contribute to a genetically informed approach to estimating risk associated with exposure to neurotoxicants. !