Eating worms: an unusual nicotinic acetylcholine receptor from the nematode pharynx as a potential drug target

吃蠕虫：线虫咽部的一种不寻常的烟碱乙酰胆碱受体作为潜在的药物靶点

基本信息

批准号：
9018496
负责人：
Alan P Robertson
金额：
$ 21.04万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-08 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9018496
关键词：
Adult Affect Age Agonist Animals Anthelmintics Ascariasis Ascaris Ascaris suum Caenorhabditis elegans Child Cholinergic Agents Cholinergic Receptors Cholinomimetics Development Disease Drosophila acetylcholine receptor alpha-subunit Drug Targeting Drug resistance Drug usage Eating Genes Growth Helminthiasis Hookworms Human Infant Infection Integral Membrane Protein Intestines Ion Channel Knowledge Lead Levamisole Life Methods Muscle Nematoda Nicotine Nicotinic Receptors Orthologous Gene Paralysed Parasite Control Parasites Parasitic infection Parasitic nematode Pharmaceutical Preparations Pharmacology Pharyngeal structure Population Protein Subunits Proteins Pyrantel Reporting Resistance Resistance development Site Soil System Testing Transmembrane Domain Trichuris Trichuris trichiura infection Veterinary Medicine Work Xenopus oocyte avermectin chemotherapy cholinergic gene cloning interest man neglected tropical diseases novel novel therapeutic intervention public health relevance receptor receptor expression response spastic paralysis

项目摘要

DESCRIPTION (provided by applicant): The Neglected Tropical Diseases (NTDs) include the soil-transmitted helminthiases (STHs) which are caused by diverse groups of intestinal nematodes. The parasites include Ascaris, Trichuris and hookworms. These infections are common. Ascariasis for example, affects 1.4 billion people worldwide and is most common in children between the ages of 3 and 8. Control of these nematode parasites relies on an effective supply of anthelmintics. Taking veterinary medicine as an example, we know that continued use of anthelmintic compounds for mass chemotherapy will lead to drug resistance. There is a need to identify novel target sites for anthelmintic development. Once such site is the nicotinic acetylcholine receptor on the nematode pharynx. We have discovered this receptor does not respond to currently used anrthelmintic drugs. Approach: The specific aims of this application are: 1. Optimize expression of C. elegans eat-2 and eat 18 and pharmacologically characterize the resulting receptor. We will investigate optimal conditions for receptor expression and characterize the resulting receptor responses using a range of agonists, antagonists and potential allosteric modulators. 2. Clone and express A. suum eat-2 and eat-18. Are other subunits required for the pharyngeal receptor? We will identify orthologs of eat-2 and eat-18 from Ascaris suum, clone, and express them in Xenopus oocytes for characterization. Additionally we will test the hypothesis that other subunit genes are required to form the receptor. 3. Do eat-2 and eat 18 combine to form the mature receptor? We will test the novel hypothesis that eat-2 and eat-18 combine to form a functional nAChR. On completion of this project we will have expressed in a heterologous system an important new potential drug target from a parasitic nematode. We will have characterized an extremely novel nAChR (one that lacks any alpha subunits). We will have identified the genes that encode these receptors in the parasitic nematode Ascaris suum. Finally, we will have determined whether the small (71 a.a.) protein eat-18 is required in the mature ion channel receptor.

描述（由申请人提供）：被忽视的热带病（NTD）包括由多种肠道线虫引起的土壤传播的蠕虫病（STH），寄生虫包括蛔虫、鞭虫和钩虫，这些感染很常见。，影响全世界 14 亿人，最常见于 3 至 8 岁的儿童。控制这些线虫以兽医学为例，我们知道，持续使用驱虫药进行大规模化疗会导致耐药性，一旦出现这种情况，就需要确定新的驱虫药靶点。线虫咽部的烟碱乙酰胆碱受体我们发现该受体对目前使用的驱虫药没有反应。 1. 优化线虫 eat-2 和 eat 18 的表达，并对所得受体进行药理学表征。我们将研究受体表达的最佳条件，并使用一系列激动剂、拮抗剂和潜在的变构调节剂表征所得受体反应。并表达 A. suum eat-2 和 eat-18 咽部受体是否需要其他亚基？我们将从中鉴定 eat-2 和 eat-18 的直系同源物？猪蛔虫、克隆并在爪蟾卵母细胞中表达它们以进行表征。此外，我们将测试形成受体所需的其他亚基基因的假设。 3. eat-2 和 eat 18 是否结合形成成熟受体？ eat-2 和 eat-18 结合形成功能性 nAChR 的新假设在该项目完成后，我们将在异源系统中表达来自寄生线虫的重要新潜在药物靶点。我们将鉴定出寄生线虫蛔虫中编码这些受体的基因。最后，我们将确定小蛋白 eat-18 是否是必需的。成熟的离子通道受体。