"Project 3" Defining the in vivo function of ncRNAs during MHV68 latency and lymphomagenesis

“项目 3”定义 ncRNA 在 MHV68 潜伏期和淋巴瘤发生过程中的体内功能

基本信息

批准号：
10646240
负责人：
Scott A. Tibbetts
金额：
$ 29.99万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-09 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646240
关键词：
Acceleration Acquired Immunodeficiency Syndrome Animal Model B-Cell Lymphomas B-Lymphocytes Biological Biological Models Biology Cell physiology Chronic Code Collaborations Comparative Study Data Development Disease Epstein-Barr virus encoded RNA 1 Epstein-Barr virus encoded RNA 2 Event Foundations Genes Genetic Transcription Goals HIV Hematogenous Herpesviridae Human Human Herpesvirus 4 Human Herpesvirus 8 Hyperplasia Immunocompromised Host Immunologics Infection Lesion Link Lymphoma Lymphomagenesis Lymphoproliferative Disorders Malignant - descriptor Malignant Neoplasms Messenger RNA MicroRNAs Molecular Mus Nature Outcome Pathogenesis Pathway interactions Peripheral Phenotype Play Proteins RNA Regulation Reporting Repression Role Shapes Signal Pathway Site Small RNA Surface System Technology Testing Transcript Transfer RNA Translations Untranslated RNA Viral Viral Physiology Viral reservoir Virus Work chronic infection defined contribution digital experimental study gammaherpesvirus in vivo infected B cell innovation latent infection mouse model mutant novel prevent programs rational design recombinant virus small hairpin RNA tumor tumor progression tumorigenesis

项目摘要

Project Summary The transforming human gammaherpesviruses EBV and KSHV establish stable latent infections in B cells, providing a lifelong reservoir of virus that can contribute to the development of malignant disease. Thus, defining the mechanisms that govern long-term latency and tumorigenesis is critical for designing rational strategies to prevent disease. In vivo studies of gammaherpesviruses in humans have been severely limited by the difficulties of working in the natural host. Murine gammaherpesvirus 68 (MHV68) is related to EBV and KSHV and causes lymphomas and lymphoproliferative disease in mice, providing a readily manipulatable small animal model for mechanistic studies of the virus/host relationship in vivo. Like EBV and KSHV, MHV68 expresses multiple forms of ncRNAs whose functions during infection and pathogenesis are largely unknown. In contrast to most gammaherpesvirus protein-coding genes, gammaherpesvirus ncRNAs are abundantly expressed in vivo during chronic infection and in hyperplastic lesions during lymphoproliferative disease, suggesting that these ncRNAs may play key conserved roles in latency and tumorigenesis. In support of this, we have demonstrated that MHV68 TMER4 and EBV EBER1 share a conserved function in hematogenous dissemination, and that in vivo suppression of a conserved miRNA target promotes B cell latency. Further, our new preliminary findings implicate both small ncRNAs and miRNAs in the genesis and progression of virus-induced B cell lymphoma. Thus, we hypothesize that gammaherpesviruses utilize a broad array of ncRNAs to shape critical B cell signaling pathways and thereby promote virus dissemination, latency and tumorigenesis. In Aim 1 (in collaboration with Project 2 and with support from Cores C and D), we will define the mechanisms by which gammaherpesvirus small RNAs promote the egress and dissemination of infected B cells, facilitate B cell latency at peripheral sites, and drive key stages of lymphomagenesis. In Aim 2 (in collaboration with Projects 2 and 3, and with support from Cores B, C and D) we will define the contribution of gammaherpesvirus miRNA repression of host mRNAs and lncRNAs to B cell latency and lymphomagenesis. The highly collaborative nature of this program, along with the systematic in vivo analyses of ncRNA mutants and recombinant viruses carrying EBV or KSHV genes, provides an extremely powerful means to determine the specific molecular mechanisms by which ncRNAs contribute to gammaherpesvirus latency and tumorigenesis.

项目概要转化的人类伽马疱疹病毒 EBV 和 KSHV 在 B 细胞中建立稳定的潜伏感染，提供可能导致恶性疾病发展的终生病毒库。因此，定义控制长期潜伏期和肿瘤发生的机制对于设计合理的策略至关重要预防疾病。人类伽玛疱疹病毒的体内研究因困难而受到严重限制在自然宿主中工作。鼠伽马疱疹病毒 68 (MHV68) 与 EBV 和 KSHV 相关并引起小鼠淋巴瘤和淋巴增殖性疾病，为研究提供了易于操作的小动物模型体内病毒/宿主关系的机制研究。与 EBV 和 KSHV 一样，MHV68 表达多种形式 ncRNA 在感染和发病机制中的功能很大程度上未知。与大多数人相比伽马疱疹病毒蛋白编码基因、伽马疱疹病毒 ncRNA 在体内大量表达慢性感染和淋巴增殖性疾病期间的增生性病变，表明这些 ncRNA 可能在潜伏期和肿瘤发生中发挥关键的保守作用。为了支持这一点，我们已经证明了 MHV68 TMER4 和 EBV EBER1 在血行传播中具有保守的功能，并且在体内抑制保守的 miRNA 靶点会促进 B 细胞潜伏期。此外，我们的新初步发现表明小 ncRNA 和 miRNA 在病毒诱导的 B 细胞淋巴瘤的发生和进展中的作用。因此，我们假设伽马疱疹病毒利用广泛的 ncRNA 来塑造关键的 B 细胞信号通路从而促进病毒传播、潜伏和肿瘤发生。目标 1（与项目 2 合作在核心 C 和 D 的支持下，我们将定义伽玛疱疹病毒小 RNA 的机制促进受感染 B 细胞的流出和传播，促进 B 细胞在周围部位的潜伏期，并驱动淋巴瘤发生的关键阶段。目标 2（与项目 2 和 3 合作，并得到 Cores 的支持） B、C 和 D) 我们将定义伽马疱疹病毒 miRNA 对宿主 mRNA 和 lncRNA 抑制的贡献 B 细胞潜伏期和淋巴瘤发生。该计划的高度协作性以及系统性对携带 EBV 或 KSHV 基因的 ncRNA 突变体和重组病毒进行体内分析，提供了确定 ncRNA 贡献的特定分子机制的极其强大的方法伽马疱疹病毒潜伏期和肿瘤发生。