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

用于对神经元细胞的可变毒性作用进行遗传分析的干细胞组

• 批准号: 8620470
• 负责人: Ted Choi
• 金额: $ 22.49万
• 依托单位: PREDICTIVE BIOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8620470
• 关键词: Affect; Animal Model; Biological Assay; Biology; Cell Line; Cells; Chromosome Mapping; Colony-Forming Units Assay; Data; Dose; ES Cell Line; Evaluation; Exhibits; Exposure to; Follow-Up Studies; Genes; Genetic; Genetic Screening; Government; Head; Health; Human; In Vitro; Life; Luciferases; Maps; Measures; Mediating; Methods; Modeling; Mus; Neuronal Differentiation; Neurons; Neurotoxins; Pathway interactions; Persons; Phase; Reporter; Resources; Risk; Risk Estimate; Severities; Stem cells; Technology; Toxic effect; Toxicant exposure; assay development; cellular engineering; cost; design; developmental neurotoxicity; embryonic stem cell; genetic analysis; homologous recombination; improved; in vitro Assay; in vivo; innovation; nerve stem cell; novel; public health relevance; response; screening; toxicant; vector

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. !

我们建议开发高通量的体外测定法以鉴定遗传因素 影响毒性对神经元分化的影响。这个项目使用一种新颖的遗传 在预测生物学上开发的映射资源，多样性杂种（DO）鼠标 ES细胞面板。使用强大的ES细胞工程方法，我们将合并神经元 分化记者做ES线。神经元干细胞（NSC）系将得出 从修改后的DO ES线的面板中，NSC面板将用于量化 毒性暴露后的神经元分化。这些数据将用于映射基因座和 鉴定介导对神经毒性反应的可变反应的基因。项目设计 优先考虑快速，敏感且高度可扩展的测定技术，并具有盒式磁带 矢量设计方法以促进构建多个测定面板以跟踪 毒性对特定类型或神经元和神经元细胞类别的影响。这 技术将使大量化合物以HTS量表进行初级筛选， 并且对于在体外和体内进行后续研究的化合物的优先级将非常有用 使用小鼠和人类细胞以及活动物模型。基因和 使用该技术确定的毒理学途径将最终改善人类 通过提供对人类发展的更详细的机械理解来进行健康 神经毒性，并将有助于估计风险的遗传知情方法 与暴露于神经毒性剂有关。 呢