Interactions between trichomonasviruses, their parasite host, and the human superhost

滴虫病毒、其寄生虫宿主和人类超级宿主之间的相互作用

基本信息

批准号：
10671988
负责人：
carrie ann Hetzel
金额：
$ 4.19万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10671988
关键词：
Adherence Biological Models Candidate Disease Gene Cells Cervical Clinical Coculture Techniques Daughter Disease Progression Double Stranded RNA Virus Double-Stranded RNA Epithelial Cells Exonuclease Gene Expression Genes Genetic High-Throughput RNA Sequencing Human IRF3 gene Infection Inflammation Inflammatory Inflammatory Response Interferons Life Cycle Stages Longitudinal Studies Malignant neoplasm of prostate Measures Mediating Morphology Mothers NF-kappa B Parasites Parents Pathway interactions Pelvis Predisposition Premature Birth Proliferating Proteins RNA RNA Viruses RNA-Binding Proteins Reporting Role Severity of illness Sexually Transmitted Diseases TLR3 gene Testing Tissue-Specific Gene Expression Trichomonas Infections Trichomonas vaginalis Vagina Viral Virion Virulence Virus Virus Diseases Virus Replication Work differential expression experimental study extracellular fitness helicase human disease human pathogen nucleoside analog response transcriptome sequencing transmission process urogenital tract virus host interaction

项目摘要

PROJECT SUMMARY Trichomonas vaginalis viruses (TVVs) are double stranded RNA viruses that persist in the obligate human parasite Trichomonas vaginalis (Tvag). Tvag is the causative agent of the human disease trichomoniasis, the most common nonviral sexually transmitted infection worldwide. Infection with TVV-containing parasites is thought to increase the severity of this disease and its complications by increasing the pro-inflammatory response by the human superhost. There are five known species in genus Trichomonasvirus (TVV1, TVV2, TVV3, TVV4, and TVV5), but most experiments to date have been performed using purified TVV1 virions or TVV1-containing isolates, due in part to the greater availability of Tvag isolates singly infected with TVV1. TVVs do not have an extracellular lifecycle and instead are transmitted vertically as the parasite divides. Further, multiple TVV species can coinfect a singular trichomonad. Thus, studying the effect of TVVs on the parasite and the human superhost has been difficult, as uninfected trichomonads cannot be infected to create isogenic TVV+ vs. TVV- strains. Using the nucleoside analog 2’-C-methylcytidine (2CMC), which has been recently reported to clear trichomonads of TVV-infection, we have generated isogenic pairs of cured and uncured strains generated from the same parent isolate. With these isogenic pairs, we propose to test our hypothesis that T. vaginalis maintains a persistent viral infection, which confers a fitness advantage to the protozoan and increases virulence to the human superhost. Thus far, we have performed high throughput RNA-sequencing with one such isogenic pair to measure differential gene expression. In doing so, we have identified several candidate genes that may be involved in limiting viral replication to tolerable levels in order to maintain long-term viral persistence. These differentially expressed genes have largely not been characterized, but some encode factors with homology to known RNA exonucleases and helicases, which we hypothesize might be involved in limiting viral replication. In Aim 1 of this project, we propose to further validate these results in additional isogenic pairs in order to define factors Tvag uses to maintain a stable trichomonasvirus infection. In Aim 2, we propose to determine impacts of TVV infection on Tvag proliferation, morphology, and adherence to human cells. We hypothesize that TVVs will increase parasite survival and attachment, providing an evolutionary explanation for long-term TVV persistence. Finally, in Aim 3, we propose to compare responses of human cells to infection with TVV+ and TVV- trichomonads across TVV species. We anticipate that TVV+ Tvag strains, regardless of virus species, will elicit stronger inflammatory responses than TVV- strains. Through this work, we hope to uncover valuable information about an important human pathogen and an intriguing model system to study virus–host interactions.

项目概要阴道毛滴虫病毒 (TVV) 是一种双链 RNA 病毒，持续存在于人类体内寄生虫阴道毛滴虫 (Tvag) 是人类疾病滴虫病的病原体。全世界最常见的非病毒性传播感染是含有 TVV 的寄生虫感染。人们认为通过增加促炎性物质来增加这种疾病及其并发症的严重程度人类超级宿主的反应毛滴虫病毒属有五种已知物种（TVV1、TVV2、 TVV3、TVV4 和 TVV5），但迄今为止大多数实验都是使用纯化的 TVV1 病毒粒子或含有 TVV1 的分离株，部分原因是单独感染 TVV1 的 Tvag 分离株更容易获得。没有细胞外生命周期，而是在寄生虫分裂时垂直传播。多种 TVV 物种可以共同感染单一滴虫，因此，研究 TVV 对寄生虫的影响。人类超级宿主一直很困难，因为未感染的毛滴虫无法被感染以产生同基因 TVV+ 与 TVV- 菌株相比，使用核苷类似物 2’-C-甲基胞苷 (2CMC)，最近有报道称为了清除 TVV 感染的滴虫，我们生成了已治愈和未治愈菌株的同基因对通过这些同基因对，我们建议检验阴道毛滴虫的假设。维持持续的病毒感染，这赋予原生动物适应性优势并增加迄今为止，我们已经用一种病毒进行了高通量 RNA 测序。这样的同基因对来测量差异基因表达在此过程中，我们已经确定了几个候选者。可能涉及将病毒复制限制在可耐受水平以维持长期病毒复制的基因这些差异表达的基因在很大程度上尚未被表征，但有一些编码因子。与已知的 RNA 核酸外切酶和解旋酶具有同源性，我们捕获的这些酶可能参与限制在该项目的目标 1 中，我们建议在其他同基因对中进一步验证这些结果。为了确定 Tvag 用于维持稳定滴虫病毒感染的因素，我们建议：确定 TVV 感染对 Tvag 增殖、形态和对人体细胞粘附的影响。保持 TVV 会增加寄生虫的存活率和依附性，这为以下现象提供了进化解释：最后，在目标 3 中，我们建议比较人类细胞对感染的反应。 TVV+ 和 TVV- 毛滴虫跨 TVV 物种，无论病毒如何，我们预计 TVV+ Tvag 菌株均存在。我们希望通过这项工作发现比 TVV 菌株更强烈的炎症反应。有关重要人类病原体的宝贵信息和研究病毒宿主的有趣模型系统互动。