ANEUGEN SCREENING IN OPTIMIZED C. ELEGANS STRAINS

虽然我们知道非整倍体在许多人类健康问题中起着重要作用，如出生缺陷和癌症，但我们对许多化学品和药物可能的非优生性知之甚少。本研究旨在建立一种快速、经济的秀丽隐杆线虫毒理检测方法。我们的实验将利用蠕虫使用X染色体不分离产生XO雄性。使用男性特异性GFP表达，我们应该能够快速和容易地评估非分离频率。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种已知的突变原，以衡量我们基于秀丽隐杆线虫的试验与哺乳动物试验之间的相似性。最后，我们将尝试使用渗透剂、P-gp转运蛋白抑制剂和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秀丽隐杆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各种秀丽隐杆线虫毒性试验的发展。拟议的商业应用：本文提出的基于秀丽隐杆线虫的非优生分析将通过以下途径实现商业化：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)用于调查化学组合，以确定保险公司基于潜在的非优生责任风险。秀丽隐杆线虫菌株的开发优化毒性测试可能有更大的商业影响，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.