ANEUGEN SCREENING IN OPTIMIZED C. ELEGANS STRAINS

虽然众所周知，非整倍体在许多人类健康问题中发挥着重要作用，如出生缺陷和癌症，但我们对许多化学品和药物可能的非整倍体知之甚少。这项建议的目的是开发一种快速、经济的线虫非整倍体检测方法，优化了毒性测试。我们的分析将利用蠕虫对X染色体不分离的利用来产生XO雄性。利用男性特异的GFP表达，我们应该能够快速和容易地评估不分离频率。在我们的实验中，我们将首先描述GFP表达和不分离之间的关系。然后，我们将测试20个已知的异常基因，以衡量我们基于线虫的测试与哺乳动物测试之间的相似性。最后，我们将尝试使用渗透剂、P-gp转运蛋白的抑制剂以及影响摄食行为和染色体分离的突变来提高检测的灵敏度。由于许多这些修饰将增加线虫对抗原以外的毒素的敏感性，我们预测我们的优化策略将促进基于线虫的各种毒性测试的发展。拟议的商业应用：这里提出的基于线虫的厌氧核素检测将通过1)应用于非常早期的药物开发，2)应用于具有非优生活性的药物的重新设计，以及3)用于调查保险公司潜在的基于非优生的责任暴露的化学组合，从而实现商业化。开发针对毒性测试进行优化的线虫菌株可能会产生更大的商业影响，因为我们计划使用这里研究的优化策略来开发广泛的毒性测试。

Although aneuploidy is known to play a significant role in many human health problems such as birth defects and cancer, we know little the about the possible aneugenicity of many chemicals and drugs. This proposal is aimed at developing a rapid, economical aneugenicity assay in C. elegans strains optimized for toxicity testing. Our assay will take advantage of the worm's use of X chromosome nondisjunction to produce XO males. Using male-specific GFP expression, we should be able to assess nondisjunction frequencies rapidly and easily. We will first characterize the relationship between GFP expression and nondisjunction in our assay. We will then test 20 known aneugens to gauge the similarity between our C. elegans-based assay and mammalian tests. Finally, we will attempt to increase the sensitivity of the assay using permeabilizing agents, inhibitors of P-gp transporters, and mutations that affect feeding behavior and chromosome segregation. Because many of these modifications will increase the sensitivity of C. elegans to toxins other than antigens, we predict that our optimization strategies will facilitate the development of a variety of C. elegans-based toxicity assays. PROPOSED COMMERCIAL APPLICATIONS: The C. elegans-based aneugen assays proposed here will be commercialized through 1) application to very early stage drug development, 2) application to the redesign of drugs with aneugenic activities, and 3) use in surveying chemical portfolios for potential aneugenicity-based liability exposure by insurers. The development of C. elegans strains optimized for toxicity testing may have an even larger commercial impact in that we plan to use the optimization strategies studied here to develop a wide range of toxicity tests.