Development of a Diversity Outcross ES Cell Panel for In Vitro Genetics of Toxico

描述（由申请方提供）：菌株背景可强烈影响动物毒性试验的结果。时间和成本的限制排除了在体内的方法，长期关注的缺乏遗传多样性的动物模型。体外平台对于理解遗传背景在毒理学测试中的作用要有效得多，并且不会牺牲任何动物。在这个项目中，我们建议建立一个大型的基因多样性小组， 来自小鼠的多样性异交系的ES细胞。DO小鼠是用于协作杂交的8个创始品系的高级杂交。DO菌株捕获实验室菌株中的几乎所有序列变体，并且比F2小鼠具有数百至数千个重组断点。它们非常适合于促进复杂的性状映射。ES细胞具有巨大的灵活性，也是体外遗传学平台的理想选择。 DO ES细胞系使DO基因组永生化，创造了可再生资源。原则上，ES细胞可以通过定向分化提供几乎任何细胞类型的途径。通过这种方式，DO ES板作为永久性的可再生资源具有非常多的用途。在第一阶段，我们将通过衍生90个独立的、经验证的DO ES系来确定可行性，并估计在第一阶段小组中观察到的15种模型毒物化合物的细胞毒性的遗传成分。在第二阶段，我们将完成600 ES线面板，确定具有显着的遗传成分的细胞毒性反应使用1400参考化合物集的化合物，并映射基因座介导的遗传成分在确定的化合物。 公共卫生相关性：人们对遗传学如何影响毒理学有着长期的兴趣，但没有成本效益高、高度可扩展的工具来研究这种关系。该项目将通过开发高通量体外平台来研究对有毒物质暴露的可变反应的遗传基础，从而改善人类健康并降低环境有毒物质的风险。

DESCRIPTION (provided by applicant): Strain background can strongly influence the outcome of toxicity tests in animals. Time and cost constraints preclude in vivo approaches to longstanding concerns about the lack of genetic diversity in animal models. An in vitro platform is vastly more efficient for understanding the role of genetic background in toxicology testing, and sacrifices no animals. In this project we propose to build a large panel of genetically diverse ES cells from the Diversity Outcross line of mice. The DO mice are an advanced intercross of the 8 founder strains used for the Collaborative Cross. The DO strains capture almost all the sequence variants in laboratory strains, and harbor hundreds to thousands more recombinational breakpoints than F2 mice. They ideally suited for facilitate complex trait mapping. ES cells offer tremendous flexibility and is also ideal for an in vitro genetics platform. DO ES cell lines immortalize DO genomes, creating a renewable resource. In principle, ES cells can provide access to almost any cell type by directed differentiation. In this way, the DO ES panel is extremely versatile as a permanent, renewable resource. In Phase I, we will establish feasibility by deriving 90 independent, validated DO ES lines and estimate the genetic component of cytotoxicity observed for 15 model toxicant compounds in the Phase I panel. In Phase II we will complete the 600 ES line panel, identify compounds with a significant genetic component to cytotoxic response using a 1400 reference compound set, and map the loci mediating that genetic component in the identified compounds. PUBLIC HEALTH RELEVANCE: There is a longstanding interest in how genetics impacts toxicology, but cost effective, highly scalable tools to investigate this relationship were not available. This project will improve human health and reduce the risk of environmental toxicants by developing a high throughput in vitro platform for investigating the genetic basis of variable response to toxicant exposure.