A Diversity Outbred ES Cell Panel for In Vitro Genetics of Toxicology

用于毒理学体外遗传学的多样性远交 ES 细胞组

• 批准号: 8920575
• 负责人: Ted Choi
• 金额: $ 49.13万
• 依托单位: PREDICTIVE BIOLOGY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-24 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8920575
• 关键词: Adoption; Aneuploidy; Animal Model; Animals; Biological Assay; Biology; Candidate Disease Gene; Cell Line; Cells; Chromosome Mapping; Complex; Computer software; Data; Data Set; Dose; ES Cell Line; Equilibrium; Evaluation; Female; Genes; Genetic; Genetic Variation; Genome; Genotype; Government; Haplotypes; Head; Health; Human; In Vitro; Laboratories; Maps; Mediating; Modeling; Mus; Outcome; Persons; Phase; Pluripotent Stem Cells; Predisposition; QTL Genes; Quantitative Trait Loci; Resources; Risk; Role; Technology; Testing; Time; Toxic Environmental Substances; Toxic effect; Toxicant exposure; Toxicity Tests; Toxicogenetics; Toxicology; Variant; base; cell type; cost; cost effective; cytotoxicity; embryonic stem cell; genetic analysis; improved; in vivo; induced pluripotent stem cell; innovation; interest; male; novel; response; screening; sex; tool; toxicant; trait

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 over 90% of all sequence variants present in laboratory strains and harbor . They are ideally suited for complex trait mapping. ES cells offer tremendous flexibililty, 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 demonstrated feasibility by deriving a small set of 100 ES cell lines and identifying a cytotoxicity QTL by in vitro genetic analysis. Since then, we have derived an additional 1,002 DO ES cell lines, and in Phase II we will validate and define screening panels of male and female DO ES cell lines that are well powered for QTL mapping. We will biologically test candidate genes to identify at least 10 mouse QTL genes, and then determine the fraction of those genes that are also modifers of cytotoxicity in human IPS cells. The key challenge to broad adoption of our technology is a demostration of the relevance of mouse QTL genes to human toxicological response. The aims of this project are not primarily directed toward building the DO ES cell lines. They are directed to validating their utility toward understanding mechanisms underlying variable susceptibility to toxicants in human cells. While there are many cell types and toxicities that can be approached using our technology, we focused this project on cytotoxicity in pluripotent stem cells to efficiently generate a well powered dataset to validat our technology. The Phase II project will establish our in vitro genetics platform as an important and novel advance in understanding human toxicological response.

描述（由申请人提供）：应变背景可以强烈影响动物毒性测试的结果。时间和成本的限制排除了体内方法，无法长期担心动物模型中缺乏遗传多样性。体外平台对于理解遗传背景在毒理学测试中的作用和不牺牲动物的作用更为有效。在这个项目中，我们建议建立一个大量的遗传多样性小组 来自小鼠多样性露出线的ES细胞。 DO MICE是用于协作交叉的8个创始人菌株的高级间变。 DO菌株捕获了实验室应变和港口中所有序列变体的90％以上。它们非常适合复杂的特质映射。 ES细胞提供了巨大的屈肌，也是体外遗传学平台的理想选择。 ES细胞系使DO基因组永生，创建可再生资源。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访问。在这个 方式，Do ES面板作为永久可再生资源非常通用。在第一阶段，我们通过得出一小部分100 ES细胞系并通过体外遗传分析鉴定细胞毒性QTL来证明可行性。从那以后，我们得出了ES细胞系的另外1,002个，在第二阶段，我们将验证和定义男性和雌性的筛选面板，可用于QTL映射的功能很好。我们将在生物学上测试候选基因，以鉴定至少10个小鼠QTL基因，然后确定那些在人IPS细胞中也是细胞毒性修饰剂的基因的分数。广泛采用我们技术的关键挑战是将小鼠QTL基因与人毒理学反应的相关性的解释。该项目的目的不是主要用于构建Do ES细胞系。他们指示验证其对人类细胞中有毒物质易感性基础机制的实用性。尽管可以使用我们的技术进行许多细胞类型和毒性，但我们将该项目集中在多能干细胞中的细胞毒性上，以有效地生成功能良好的数据集以验证我们的技术。 II期项目将建立我们的体外遗传学平台，作为了解人类毒理学反应的重要而新颖的进步。