Functional Genomic Approach to Macrofilaricide Discovery

描述（由申请人提供）： 河盲症和淋巴丝虫病是两种由丝虫引起的疾病，属于最重要的热带疾病。线虫盘尾丝虫（Onchocercus volvulus）的黑蝇传播导致河盲症（盘尾丝虫病），而蚊子传播的丝虫（包括班氏丝虫（Wucheria bancrofti）和马来丝虫（Brugia malayi））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。为控制这些疾病而进行的全球协调努力集中于病媒控制和使用现有药物疗法。然而，目前的药物主要针对微丝蚴阶段，对长寿的成虫线虫无效。此外，最近还记录了对当前疗法的耐药性。尽管目前作出了努力，但根除这些疾病仍将极具挑战性。需要针对寄生虫的微丝蚴（成虫）阶段的新类别的驱虫药。在这项研究中，最近产生的线虫基因组数据将筛选与生物信息学过滤器中的马来丝虫线虫的潜在靶基因的识别。B。malayi C.将选择具有严重RNAi表型的线虫基因，以及寄生线虫特有的许多基因。将从B克隆指定基因。Malayi，并测试在成年雌性中的表达。在确认成虫表达后，将选择10 - 15个基因靶标并通过RNAi测试对成虫B生物学的影响。马来雌虫体外培养。将监测微丝蚴的存活、形态和释放，以及通过RT-PCR监测靶基因转录水平。这项工作的结果将是选择和排名的数量有限的高优先级的目标macrofilaricide药物发现。

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.