A high throughput screen for inhibitors of nematode detoxification genes

线虫寄生在约25%的人类中。蠕虫靶向药物，或驱虫药， 几十年来，人们一直在用它来控制寄生线虫，许多物种正在进化出多药耐药性。在 在多种生物体中，多药耐药性是由酶的表达和活性增加介导的， 使异生物质解毒。靶向异生物质解毒途径的药物化合物将提供 研究多药耐药性所急需的工具，可以大大延长现有的 未来的驱虫药很少有药理学化合物可用于研究和靶向多药 抗性和可用的抗性不是线虫特异性的，并且仅针对单一酶或类别 酶。 转录因子SKN-1激活外源性解毒基因的表达， 线虫线虫SKN-1基因的抑制使C.到各种异生物质， 和C.线虫可以通过增加基因的表达来获得对驱虫剂的抗性， 由SKN-1调节。SKN-1对胚胎发育也至关重要。我们最近的研究 确定了一个调节SKN-1的主要途径，该途径与调节外源性物质的途径高度不同。 哺乳动物的解毒因此，SKN-1是一个很有前途的靶点，可以用来开发药物， 胚胎发育，降低胁迫抗性，并抑制线虫的异生物质解毒， 影响人类的类似途径。其个体小、养殖简单、遗传多样性强， C.易处理性elegans使其成为筛选异生物质解毒基因抑制剂的理想系统。 该项目的第一个目标是开发和优化SKN-1活性的荧光检测方法， C.优雅该项目的第二个目标是开发四个二级和计数器屏幕， 对命中化合物进行优先级排序，并在2000种化合物的中试筛选中测试该测定法。在完成这些 目标，将提交优化测定的详细描述以输入分子库 探针生产中心网络（MLPCN）。

Nematodes parasitize ~25% of the human population. Helminth targeting drugs, or anthelmintics, have been used to control parasitic nematodes for decades and many species are evolving multidrug resistance. In diverse organisms, multidrug resistance is mediated by increased expression and activity of enzymes that detoxify xenobiotics. Pharmacological compounds that target xenobiotic detoxification pathways would provide much needed tools for studying multidrug resistance and could greatly increase the useful life of current and future anthelmintics. Few pharmacological compounds are available for studying and targeting multidrug resistance and those that are available are not specific for nematodes and only target a single enzyme or class of enzymes. The transcription factor SKN-1 activates the expression of xenobiotic detoxification genes in the nematode Caenorhabditis elegans. Genetic inhibition of SKN-1 sensitizes C. elegans to diverse xenobiotics, and C. elegans can acquire resistance to anthelmintics by increasing the expression of genes that are regulated by SKN-1. SKN-1 is also essential for the development of embryos. Our recent studies have identified a principal pathway regulating SKN-1 that is highly divergent from pathways that regulate xenobiotic detoxification in mammals. Therefore, SKN-1 is a promising target for the development of drugs that disrupt embryonic development, decrease stress resistance, and inhibit xenobiotic detoxification in nematodes without affecting analogous pathways in humans. The small size, simple culturing characteristics, and genetic tractability of C. elegans makes it an ideal system to screen for inhibitors of xenobiotic detoxification genes. The first goal of this project is to develop and optimize a fluorescence-based assay of SKN-1 activity in C. elegans. The second goal of this project is to develop four secondary and counter screens to rapidly prioritize hit compounds and to test the assay in a pilot screen of 2000 compounds. After completion of these goals, a detailed description of the optimized assay will be submitted for entry into the Molecular Libraries Probe Production Centers Network (MLPCN).