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（由申请方提供）：线虫寄生于约25%的人群。蠕虫靶向药物或驱虫剂已用于控制寄生线虫数十年，许多物种正在发展多药耐药性。在不同的生物体中，多药耐药性是由增加的表达和活性的酶，解毒异生物质介导的。靶向外源性解毒途径的药物化合物将为研究多药耐药性提供急需的工具，并可大大延长当前和未来驱虫剂的使用寿命。很少有药理学化合物可用于研究和靶向多药耐药性，并且那些可用的化合物不是线虫特异性的，并且仅靶向单一酶或一类酶。 转录因子SKN-1激活线虫中异生物质解毒基因的表达。SKN-1基因的抑制使C. elegans和C.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剂的抗性。SKN-1对胚胎发育也至关重要。我们最近的研究已经确定了一个主要的途径调节SKN-1是高度不同的途径，调节外源性解毒哺乳动物。因此，SKN-1是一个很有前途的目标，用于开发药物，破坏胚胎发育，降低应激抵抗力，并抑制线虫中的异生物质解毒，而不影响人类中的类似途径。该菌体积小，培养简单，遗传易控制。线虫使其成为筛选异生物质解毒基因抑制剂的理想系统。 本项目的第一个目标是开发和优化一种基于荧光的检测C.优雅的该项目的第二个目标是开发四个二级和反筛选，以快速优先考虑命中化合物，并在2000种化合物的中试筛选中测试该测定法。完成这些目标后，将提交优化检测的详细描述，以进入分子库探针生产中心网络（MLPCN）。 公共卫生相关性：线虫寄生于约25%的人类，许多菌株对目前使用的所有抗寄生虫药物都具有耐药性。在许多生物体中，耐药性是由过度活跃的药物解毒途径引起的。本申请中提出的研究将开发、优化和验证线虫药物解毒的药理学抑制剂的筛选。