Genetically Diverse Embryonic Stem Cell Lines For Reproductive Toxicology

说明(申请人提供)：确定化合物的生殖毒性风险对于确保普通人群的健康至关重要。这是一项困难而复杂的任务，涉及到动物试验。虽然众所周知，小鼠品系背景可以对表型产生深远的影响，但理解品系背景在毒性测试中的作用仍然是一个巨大的挑战。体外基因测试系统通过提供更低的成本、更高的吞吐量和在测试中不牺牲动物的优势为这一目标做出贡献。体外遗传系统可以作为“第一层”测试平台，指导对小鼠品系的体内测试，从而最大限度地增加信息量，减少动物使用。我们建议开发一种基于ES细胞的体外系统，以评估毒性测试中遗传背景的影响。ES细胞对于这一目的特别有吸引力，因为它们可以在其全能状态下无限繁殖，同时保留对动物所有组织做出贡献的能力[3]。在第一阶段，我们将通过从少数遗传差异的小鼠品系建立ES细胞系，并使用参考化合物在胚胎干细胞测试(EST)中对它们进行评估，来确定该系统的可行性。在第二阶段，将建立更多的ES品系，并用更大的参考化合物小组进行测试，以开发一个系统来定义EST中毒性的遗传成分。该系统将通过提供一个平台来调查遗传背景对毒性风险的作用，从而导致更具预测性的生殖毒性测试。该测试平台将向制药和化工行业以及学术机构提供商业使用。我们建议开发一种系统，帮助阐明遗传学在生殖毒性测试中的作用。拟议检测系统中广泛而多样的遗传背景更好地反映了美国人口的遗传多样性，并可能导致更具预测性的测试，从而降低对人口的环境健康风险。

DESCRIPTION (provided by applicant): Determining the reproductive toxicity risk of compounds is vitally important to ensure the health of the general population. It is a difficult and complex task involving testing in animals. While it is well known that mouse strain background can have a profound effect on phenotype, understanding the role of strain background in toxicity testing remains a tremendous challenge. An in vitro genetic testing system contributes toward this end by offering the advantages of lower cost, higher throughput and no sacrifice of animals in testing. An in vitro genetic system can be a "first tier" testing platform, directing in vivo testing to mouse strains that maximize informativeness and minimize animal use. We propose to develop an in vitro, ES cell based system to assess the impact of genetic background in toxicity testing. ES cells are particularly attractive for this purpose since they can be propagated indefinitely in their pluropotent state while retaining the ability to contribute to all tissues of an animal [3]. In Phase I we will determine the feasibility of this system by establishing ES cell lines from a small number of genetically distinct mouse strains and evaluating them in the Embryonic Stem Cell Test (EST) with a reference compound. In Phase II, additional ES lines will be established and tested with a larger panel of reference compounds, toward the development of a system to define the genetic components of toxicity in the EST. This system will lead to more predictive reproductive toxicity testing by providing a platform to investigate the role of genetic background on toxicity risk. The testing platform will be made commercially available to the pharmaceutical and chemical industries, as well as to academic institutions. We propose to develop a system that can help elucidate the role of genetics in reproductive toxicity testing. The broad and varied genetic backgrounds in the proposed testing system better reflects the genetic diversity of the US population, and may lead to more predictive testing that reduces environmental health risk to the population.