Exploring the Structure and Function of Parasitic Nematode Exosomes

淋巴丝虫病（LF）是一种由蠕虫引起的蚊媒被忽视的热带疾病， 马来丝虫;全世界有超过1.2亿人受到感染，在70个地方性流行病中有超过14亿人处于危险之中。 国家目前采用大规模药物管理的控制战略在许多国家降低了流行率， LF仍然是一个重大的全球健康问题。人们认识到需要制定新的战略， 防治LF等寄生线虫引起的病害。这个项目的重点是宿主-寄生虫 界面寄生虫感染，发展和持续的LF，并代表了一个令人兴奋的新方向 来调查这个领域我们提出了一种新的机制，其中B。马来人调节宿主免疫 系统，通过小的，调节RNA和蛋白质通过一种特定类型的细胞外囊泡传递，称为 外来体我们的初步数据表明，感染期幼虫的B。马来人分泌外泌体， 这些外来体含有不同的miRNA和蛋白质货物，不同的寄生虫miRNA可能靶向 宿主基因此外，这些外泌体被宿主巨噬细胞内化，并引起特异性调节。 表型该提案的总体目标是定义跨膜分泌的寄生虫外泌体的货物。 哺乳动物内的布氏丝虫的生命周期，并探讨其生物活性的机制基础。 在具体目标1中，我们建议分析分泌的外泌体的小RNA和蛋白质货物， 哺乳动物内B.使用小RNA深度测序（RNA-Seq）和 蛋白质组分析，以确定由寄生虫外泌体递送的宿主操纵的分子介质。在 具体目标2，我们将定义外泌体生物活性对宿主免疫的细胞介质的机制， 反应我们将研究宿主巨噬细胞对寄生虫外泌体的摄取，以揭示这些囊泡如何 被宿主细胞内化，然后通过使用基因敲除来利用小鼠模型的遗传能力， 外泌体在宿主巨噬细胞中引起的调节表型的机制 内化 该项目的长期影响将是B的新知识。马来西亚生物学和新的 分子，可以利用在新的LF控制策略。此外，我们在这里描述的机制可能 在动物、人类和植物寄生线虫中是保守的，并且可以用于广谱的 控制应用。

Lymphatic filariasis (LF) is a mosquito-borne Neglected Tropical Disease caused by filarial worms including Brugia malayi; over 120 million people worldwide are infected, with over 1.4 billion at risk in 70 endemic countries. Current control strategies employing mass drug administration have reduced prevalence in many areas, but LF remains a significant global health concern. There is a recognized need for new strategies to control LF and other diseases caused by parasitic nematodes. This project focuses on the host-parasite interface during parasite infection, development and persistence of LF and represents an exciting new direction for investigating this field. We propose a novel mechanism by which B. malayi modulates the host immune system, through small, regulatory RNAs and proteins delivered via a specific type of extracellular vesicle called exosomes. Our preliminary data demonstrate that infective stage larvae of B. malayi secrete exosomes, that these exosomes contain a diverse miRNA and protein cargo and that distinct parasite miRNA potentially target host genes. Further, these exosomes are internalized by host macrophages and elicit a specific modulatory phenotype. The overall goals of this proposal are to define the cargo of parasite exosomes secreted across the intra-mammalian Brugia life cycle and probe the mechanistic basis for their bioactivity. In Specific Aim 1, we propose to profile the small RNA and protein cargo of secreted exosomes across the intra-mammalian B. malayi life cycle using a combination of small RNA deep-sequencing (RNA-Seq) and proteomic profiling to identify the molecular mediators of host manipulation delivered by parasite exosomes. In Specific Aim 2, we will define the mechanisms of exosome bioactivity on cellular mediators of the host immune response. We will examine uptake of parasite exosomes by host macrophages to reveal how these vesicles are internalized by host cells, then leverage the genetic capacity of the murine model by using genetic knock outs to generate mechanistic insight into the modulatory phenotype elicited in host macrophages by exosome internalization. The long-term impact of the project will be new knowledge of B. malayi biology and the exposure of new molecules that may be exploited in novel LF control strategies. Further, the mechanisms we describe here may be conserved across animal, human and plant parasitic nematodes and could be utilized for broad-spectrum control applications.