Regulation of Herpesvirus Infection by Viral miRNAs

病毒 miRNA 对疱疹病毒感染的调控

基本信息

批准号：
8535928
负责人：
Linda F. Van Dyk
金额：
$ 36.8万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-15 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8535928
关键词：
Acute Address Animal Model Animals Antigen Presentation Apoptosis Attenuated B-Lymphocytes Benign Bioinformatics Biological Assay Biological Models Cell Cycle Cells Chronic Chronic Disease Defect Disease Disease Outcome Gene Expression Generations Genetic Genetic screening method Health Herpesviridae Infections Human Human Herpesvirus 4 Human Herpesvirus 8 Immune Immune system In Vitro Individual Infection Inflammatory Investigation Measures Mediating MicroRNAs Modeling Molecular Mus Mutagenesis Nucleic Acids Outcome Pathogenesis Pathway interactions Phenotype Physiologic pulse Process Protein Family Proteins Proteome Proteomics Reagent Recombinants Regulation Research Risk Factors Role Series Signal Transduction Site Small RNA Staging Support System System Testing Therapeutic Vaccines Validation Viral Viral Genes Virus Virus Diseases Virus Replication Work base cell type defined contribution disorder risk gammaherpesvirus immune function in vitro Model in vivo innovation insight latent infection mutant pathogen reactivation from latency recombinant virus restoration tumor virus identification

项目摘要

DESCRIPTION (provided by applicant): Gammaherpesviruses, including the human gHV Epstein Barr virus and Kaposi's sarcoma associated herpesvirus, infect cells of the immune system and establish lifelong, chronic infection that can exist in a benign, latent state. Decreased immune function (from congenital, acquired or iatrogenic immunedeficiencies) is a risk factor for gHV-associated diseases including tumors and chronic inflammatory disease. Important advances in our understanding of these human pathogens can be made from animal models of infection and disease. We hypothesize that these viral regulatory RNAs serve similar functions in these related gHV, much as divergent viral genes in these viruses target common molecular pathways (such as antigen presentation, cell cycle and apoptosis). We will 1) Define the activity of and requirement for the gHV68 miRNAs in vitro, 2) Test the genetic requirement of viral miRNAs to gHV68 infection and pathogenesis in vivo, and 3) Investigate and identify conserved targets of gHV miRNAs. To accomplish this, we will complete the generation and validation of several mutant viruses, lacking viral miRNAs or lacking a viral target sequence, we will make use of an enzymatically marked virus for identification and purification of infected cells, we will determine the integrated protein changes attributable to the viral miRNAs, and most importantly, we will conduct these studies in the context of whole animal infection. These studies will allow us to definitively address the genetic contribution of virally-encoded miRNAs to in vivo pathogenesis, a question uniquely addressed by the gHV68 model of gHV pathogenesis. These studies are highly likely to provide new insights into gHV miRNAs (e.g. virus alterations suitable for vaccines, identification of targeted pathways for therapeutic restoration, or identification of pathways for small RNA regulation that could be useful in other disease states).

描述（由申请人提供）：γ疱疹病毒，包括人类gHV Epstein Barr病毒和卡波西肉瘤相关疱疹病毒，感染免疫系统的细胞并建立可以以良性、潜伏状态存在的终生慢性感染。免疫功能下降（先天性、后天性或医源性免疫缺陷）是 gHV 相关疾病（包括肿瘤和慢性炎症性疾病）的危险因素。我们对这些人类病原体的理解可以从感染和疾病的动物模型中取得重要进展。我们假设这些病毒调节RNA在这些相关的gHV中发挥相似的功能，就像这些病毒中不同的病毒基因针对共同的分子途径（例如抗原呈递、细胞周期和细胞凋亡）一样。我们将1）在体外定义gHV68 miRNA的活性和需求，2）在体内测试病毒miRNA对gHV68感染和发病机制的遗传需求，以及3）研究和鉴定gHV miRNA的保守靶标。为了实现这一目标，我们将完成几种突变病毒的生成和验证，这些突变病毒缺乏病毒miRNA或缺乏病毒靶序列，我们将利用酶标记病毒来识别和纯化受感染的细胞，我们将确定整合的蛋白质变化归因于病毒 miRNA，最重要的是，我们将在整个动物感染的背景下进行这些研究。这些研究将使我们能够明确地解决病毒编码的 miRNA 对体内发病机制的遗传贡献，这是 gHV 发病机制的 gHV68 模型独特解决的问题。这些研究很可能为 gHV miRNA 提供新的见解（例如，适合疫苗的病毒改变、治疗恢复的靶向途径的鉴定，或可用于其他疾病状态的小 RNA 调节途径的鉴定）。