A high throughput screen for inhibitors of nematode detoxification genes

线虫解毒基因抑制剂的高通量筛选

• 批准号: 8000247
• 负责人: KEVIN STRANGE
• 金额: $ 11.78万
• 依托单位: MOUNT DESERT ISLAND BIOLOGICAL LAB
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8000247
• 关键词: Drug Metabolic Detoxication; Genes; Nematoda; high throughput screening; inhibitor/antagonist

DESCRIPTION (provided by applicant): 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 make 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). PUBLIC HEALTH RELEVANCE: Nematodes parasitize ~25% of humans and many strains are resistant to all currently used antiparasitic drugs. In many organisms, drug resistance is caused by over active drug detoxification pathways. The studies proposed in this application will develop, optimize, and validate a screen for pharmacological inhibitors of drug detoxification in nematodes.

描述（由申请人提供）：线虫寄生约25％的人口。数十年来，用于控制寄生线虫的蠕虫靶向药物或驱虫药已被用于控制寄生线虫，许多物种正在发展多药的耐药性。在各种生物体中，多药耐药性是由对异种生物的酶的表达和活性的增加和活性介导的。靶向异种生物排毒途径的药理学化合物将提供急需研究多药耐药性的工具，并可以大大提高当前和未来的驱虫药的使用寿命。很少有药理学化合物可用于研究和靶向多药耐药性，而可获得的化合物并非针对线虫，仅针对单个酶或一类酶。 转录因子SKN-1激活了线虫秀丽隐杆线虫中异种生物解毒基因的表达。 SKN-1的遗传抑制作用使秀丽隐杆线虫对多种异生物生物敏感，秀丽隐杆线虫可以通过增加受SKN-1调节的基因的表达来获得对驱虫药的抗性。 SKN-1对于胚胎的发展也是必不可少的。我们最近的研究确定了调节SKN-1的主要途径，该途径与调节哺乳动物中异种生物排毒的途径高度不同。因此，SKN-1是破坏胚胎发育，降低压力抗性并抑制线虫中的非生物生物解毒的药物开发的有希望的目标，而不会影响人类类似的途径。秀丽隐杆线虫的小尺寸，简单的培养特性和遗传性障碍性使其成为筛选异种生物学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秀丽隐杆线虫中SKN-1活性的基于荧光的测定法。该项目的第二个目标是开发四个二次和计数器屏幕，以快速优先考虑命中化合物，并在2000种化合物的试验屏幕中测试测定法。完成这些目标后，将提交对优化测定的详细描述，以进入分子库探测生产中心网络（MLPCN）。 公共卫生相关性：线虫寄生约25％的人类，许多菌株对当前使用的抗寄生虫药物具有抗性。在许多生物体中，耐药性是由活性药物排毒途径过多引起的。本应用中提出的研究将开发，优化和验证线虫中药物解毒的药理抑制剂的筛选。