Extracellular vesicles, small RNAs, and intercellular communication in Entamoeba histolytica

溶组织内阿米巴的细胞外囊泡、小 RNA 和细胞间通讯

• 批准号: 9165169
• 负责人: UPINDER SINGH
• 金额: $ 19.75万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-10 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9165169
• 关键词: Amoeba genus; Biochemical; Bioinformatics; Biological; Biological Process; Biology; Cause of Death; Cell Communication; Cell Line; Cell physiology; Cells; Cellular biology; Communication; Communities; Complex; Cyst; Data; Data Set; Disease; Dysentery; Enabling Factors; Entamoeba; Entamoeba histolytica; Environment; Exploratory/Developmental Grant; Gene Expression Regulation; Gene Silencing; Gene Silencing Pathway; Generations; Grant; Health; Human; Immune response; Infection; Knowledge; Libraries; Life Cycle Stages; Light; Liver Abscess; Maintenance; Mass Spectrum Analysis; Mediating; MicroRNAs; Molecular; Nematoda; Organism; Parasites; Pathogenesis; Pathway interactions; Plasmodium; Polyphosphates; Polyps; Population; Predisposition; Process; Proteins; RNA Interference; RNA library; RNA-Binding Proteins; Reagent; Regulation; Role; Small RNA; Staging; Structure; System; Tissues; Trichomonas; Virulence; Work; exosome; experience; extracellular; extracellular vesicles; genome-wide; high risk; improved; intercellular communication; interest; member; novel; pathogen; pressure; success; tool; trafficking

Entamoeba histolytica, a protozoan parasite, is an important human pathogen. Diseases caused by E. histolytica include dysentery and liver abscesses, and the organism is a leading parasitic cause of death on a global scale. Regulation of gene expression is a key factor that enables the parasite to adapt to the host environment during tissue invasion and to convert to the cyst stage and propagate disease outside the host. One mechanism of gene regulation in Entamoeba is a robust and complex endogenous RNAi pathway. Over the last several years we have made important observations about this pathway including that small RNAs associate with amebic Argonaute protein to mediate transcriptional gene silencing and have highly unusual 5'- polyP termini. However, despite extensive efforts we have not identified which aspects of amebic biology are regulated by small RNAs, although given that the pathway is maintained in related Entamoeba, a strong evolutionary pressure and clear biological role must exist. We have recently identified that ameba secrete extracellular vesicles that contain small RNAs and RNAi effector proteins. We now seek to determine whether these extracellular vesicles containing small RNAs serve as a means of host-parasite or parasite-parasite intercellular communication. Thus, we will (i) identify small RNA repertoire of amebic extracellular vesicles, (ii) determine if they participate in intercellular communication, and (iii) determine protein components of EVs with focus on RNA binding proteins. These data will improve our understanding of the molecular mechanisms of RNAi in Entamoeba and will also broaden our understanding of the roles of small RNAs in intercellular communication. Our work is at the intersection of the basic cellular process of RNA- interference and amebic biology. Data that emerge will contribute to both understanding amebic pathogenesis but also to expanding the knowledge about the fundamental process of RNAi.

溶组织内阿米巴是一种原生动物寄生虫，是人类重要的病原体。由E. 溶组织性疾病包括痢疾和肝脓肿，这种有机体是一种主要的寄生性死亡原因 全球范围。基因表达的调节是使寄生虫适应宿主的关键因素 在组织入侵期间的环境中，并转换到囊肿期，并在宿主外传播疾病。 内阿米巴基因调控的一个机制是一个强大而复杂的内源RNAi途径。完毕 在过去的几年里，我们对这一途径进行了重要的观察，包括小RNA 与阿米巴ArgAerte蛋白结合，介导转录基因沉默，并具有极不寻常的5‘- 息肉终末。然而，尽管我们做了大量的努力，我们还没有确定阿米巴生物学的哪些方面 受小RNA调控，尽管该途径是在相关的内阿米巴中维持的，但 进化压力和明确的生物学作用必须存在。我们最近发现阿米巴能分泌 含有小RNA和RNAi效应蛋白的细胞外小泡。我们现在寻求确定是否 这些含有小RNA的胞外小泡是寄主-寄生虫或寄生虫-寄生虫的一种手段 细胞间通信。因此，我们将(I)鉴定阿米巴胞外的小RNA谱系 囊泡，(Ii)确定它们是否参与细胞间通讯，以及(Iii)确定蛋白质 EVS的成分，重点是RNA结合蛋白。这些数据将提高我们对 RNAi在内阿米巴中的分子机制，也将拓宽我们对Small作用的理解 细胞间通讯中的RNA。我们的工作是在RNA的基本细胞过程的交叉点上- 干扰和阿米巴生物学。出现的数据将有助于理解阿米巴的发病机制 而且还有助于扩大对RNAi基本过程的了解。