Molecular mechanisms controlling secretion in filarial nematode parasites

项目摘要 寄生线虫感染超过15亿人。控制这种与贫困相关的全球健康负担依赖于 几乎完全依赖于少量驱虫药的施用。驱虫药的发展前景 这些药物在许多线虫寄生虫中的耐药性和次优性质需要新的方法来 寄生虫治疗和控制。然而，开发新的抗寄生虫治疗选择的需要受到阻碍 由于我们对线虫生物学过程的基础知识存在很大的差距， 维持感染。寄生线虫释放到宿主体内的排泄分泌（ES）产物 环境对于宿主免疫调节和成功寄生是必不可少的。尽管总的来说- 认为ES系统是释放促进寄生虫的分子（蛋白质和囊泡）的管道， 生存，我们对ES装置的基本结构和功能在医学上的理解很差。 重要的寄生线虫。为了解决这一知识空白，本项目将确定分泌调节因子， 在马来丝虫（一种蚊子传播的丝虫线虫和人类淋巴膜的病原体）中发挥作用， （LF）. B.马来西亚人，包括关于伊维菌素作用方式的人，支持以下假设： ES装置是新治疗靶点的一个有利可图的和未开发的来源。我们的首要假设是 是细胞表面受体定位于B。马来ES系统直接或间接控制寄生虫分泌 功能，并且它们可以被靶向以干扰ES衍生分子的释放。我们将奉行 三个目标，由初步的受体线索驱动，并通过我们优化的创新方法变得可行 解析马来B.malayi ES系统的转录组状态，并分析与寄生虫自身相关的受体， cretory函数我们专注于G蛋白偶联受体（GPCRs），这是一种重要的药物靶点 并且已知在线虫ES细胞和可作用于ES系统的邻近细胞类型中表达。在 目的1，我们将使用创新的空间转录组学方法来解析B的转录组。马来丝虫 ES区域跨宿主内阶段，并确定候选GPCR调节ES功能。在目标2中，我们 使用反向遗传学和化学方法来评估ES定位的GPCR在调节 B。马来人分泌功能在目标3中，我们将使用全生物体模型线虫和哺乳动物单细胞 异源表达平台，以定义ES定位的GPCR的药理学，并建立功能性 用于GPCR筛选的分析。该项目的完成将产生有关财务的基本新知识， 线虫ES系统，并提供新的先导目标和验证筛选新的抗疟药物发现。

Project Summary Parasitic nematodes infect over 1.5 billion humans. Control of this poverty-associated global health burden relies almost entirely on the administration of a small number of anthelmintic drugs. The prospects of anthelmintic resistance and the sub-optimal nature of these drugs in many nematode parasites demand new approaches to parasite treatment and control. However, the need to develop new antiparasitic treatment options is hampered by large gaps in our basic knowledge of the nematode biological processes that promote the establishment and maintenance of infection. Excretory-secretory (ES) products released by parasitic nematodes into their host environments are essential for host immune modulation and successful parasitism. Despite the general under- standing that the ES system is a conduit for the release of molecules (proteins and vesicles) that promote parasite survival, we have a poor understanding of the underlying structure and function of the ES apparatus in medically important parasitic nematodes. To address this gap in knowledge, this project will identify regulators of secretory function in Brugia malayi, a mosquito-transmitted filarial nematode and causative agent of human lymphatic filar- iasis (LF). Recent studies in B. malayi, including on the mode of action of ivermectin, support the premise that the ES apparatus is a lucrative and unexploited source of new therapeutic targets. Our overarching hypothesis is that cell-surface receptors localized to the B. malayi ES system directly or indirectly control parasite secretory function, and that they can be targeted to interfere with the release of ES-derived molecules. We will pursue three aims, motivated by preliminary receptor leads and made feasible by innovative methods we have optimized to resolve the transcriptomic state of the B.malayi ES system and to profile receptors implicated in parasite se- cretory function. We focus our efforts on G protein-coupled receptors (GPCRs), which are prolific drug targets and known to be expressed in nematode ES cells and adjacent cell types that may act on the ES system. In Aim 1, we will use innovative spatial transcriptomics approaches to resolve the transcriptome of the B. malayi ES region across intra-host stages and to identify candidate GPCRs that regulate ES function. In Aim 2, we will use reverse genetics and chemical approaches to assess the role of ES-localized GPCRs in the regulation of B. malayi secretory function. In Aim 3, we will use whole-organism model nematode and mammalian single-cell heterologous expression platforms to define the pharmacology of ES-localized GPCRs and to establish functional assays for GPCR screening. Completion of this project will produce fundamental new knowledge about the filarial nematode ES system and deliver new lead targets and validated screens for novel anti-filarial drug discovery.