Functional Genomic Approach to Macrofilaricide Discovery

发现大丝虫杀剂的功能基因组方法

• 批准号: 6882175
• 负责人: James P. McCarter
• 金额: $ 10万
• 依托单位: DIVERGENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-15 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882175
• 关键词: Caenorhabditis elegans; Filarioidae; RNA interference; Wuchereria; anthelmintics; antiparasitic agents; bioinformatics; drug design /synthesis /production; drug discovery /isolation; drug resistance; filariasis; functional /structural genomics; helminth genetics; parasite infection mechanism; parasitic disease diagnosis; polymerase chain reaction; transfection /expression vector

DESCRIPTION (provided by applicant): River blindness and lymphatic filariasis, two disease caused by filarial nematodes, are among the most important tropical diseases. Blackfly transmission of the nematode Onchocercus volvulus leads to river blindness (onchocerciasis), while mosquito-transmitted filarial worms including Wucheria bancrofti and Brugia malayi are responsible for lymphatic filariasis. Adult worms survive in infected individuals for many years, continuously releasing microfilarial progeny into the blood. Coordinated global efforts to control these diseases have concentrated on vector control and the use of existing drug therapies. However, the current drugs primarily target the microfilarial stages and are not effective against the long-lived adult nematodes. Additionally, drug resistance to current therapies has been recently documented. Despite current efforts, eradication of these diseases will be extremely challenging. New classes of anthelmintic drugs are needed which target the macrofilaria (adult) stages of parasitic worms. In this research, recently generated nematode genomic data will be screened with bioinformatic filters for the identification of potential target genes in the filarial nematode Brugia malayi. B. malayi orthologs of C. elegans genes with severe RNAi phenotypes will be chosen, as well as a number of genes unique to parasitic nematodes. Nominated genes will be cloned from B. malayi, and tested for expression in adult females. Following confirmation of adult expression, 10-15 gene targets will be selected and tested by RNAi for effects on the biology of adult B. malayi female worms in vitro. Survival, morphology, and release of microfilaria will be monitored, as will target gene transcript levels by RT-PCR. The outcome of this work will be the selection and ranking of a limited number of high priority targets for macrofilaricide drug discovery.

描述（由申请人提供）： 河盲症和淋巴丝虫病这两种由丝虫线虫引起的疾病是最重要的热带疾病。盘尾丝虫线虫的黑蝇传播会导致河盲症（盘尾丝虫病），而蚊子传播的丝虫（包括班氏丝虫和马来丝虫）则导致淋巴丝虫病。成虫在感染者体内存活多年，不断将微丝蚴后代释放到血液中。控制这些疾病的全球协调努力集中在病媒控制和现有药物疗法的使用上。然而，目前的药物主要针对微丝蚴阶段，对长寿命线虫成虫无效。此外，最近还记录了对当前疗法的耐药性。尽管目前做出了努力，根除这些疾病仍将极具挑战性。需要针对寄生虫的大丝虫（成虫）阶段的新型驱虫药。在这项研究中，将使用生物信息过滤器筛选最近生成的线虫基因组数据，以鉴定丝虫线虫马来丝虫中的潜在靶基因。将选择具有严重RNAi表型的线虫基因的马来芽孢杆菌直系同源物，以及寄生线虫特有的许多基因。提名基因将从马来蝽中克隆，并测试其在成年雌性中的表达。确认成虫表达后，将选择 10-15 个基因靶标，并通过 RNAi 测试其对马来双歧杆菌雌虫体外生物学的影响。将监测微丝蚴的存活、形态和释放，并通过 RT-PCR 检测目标基因转录水平。这项工作的结果将是对有限数量的杀大丝虫药物发现的高优先目标进行选择和排序。