Defining the Mechanisms of Epstein-Barr Virus Persistence and Recurrence

定义 Epstein-Barr 病毒持续存在和复发的机制

基本信息

批准号：
10599350
负责人：
Micah A. Luftig
金额：
$ 46.71万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-04 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10599350
关键词：
Adolescence Adult Apoptotic Architecture B lymphocyte immortalization B-Cell Activation B-Lymphocytes BAG3 gene Benign Biochemical Biological Biology Cell Differentiation process Cell Fate Control Cell Lineage Cell Maturation Cell Survival Cells Cellular Metabolic Process Chromatin Data Epithelial Cells Epstein-Barr Virus latency Gene Expression Genes Genetic Transcription Goals Herpesviridae Human Human Herpesvirus 4 Immune Immunoglobulin Class Switching Immunoglobulin-Secreting Cells Individual Infection Infectious Mononucleosis Latent virus infection phase Life Cycle Stages Link Lymphoid Tissue Lytic Lytic Virus MCL1 gene Malignant Neoplasms Mediating Memory Memory B-Lymphocyte Mitochondria Modality Modeling Molecular Molecular Chaperones Molecular Conformation NFKB2 gene Oral Oral cavity PRDM1 gene Phenotype Plasma Cells Plasmablast Primary Infection Productivity Proliferating Proteins Reaction Recurrence Regulation Research Rest Saliva Sampling Signal Transduction Structure of germinal center of lymph node Testing Therapeutic Tonsil Viral Viral Genes Viral Load result Virion Virus XBP1 gene YY1 Transcription Factor cell immortalization in vivo insight latent infection mimicry novel peripheral blood plasma cell differentiation programs reactivation from latency recruit single-cell RNA sequencing transcription factor transmission process virtual

项目摘要

ABSTRACT Our ultimate goal to define the molecular mechanisms for EBV latency establishment and reactivation in the oral cavity. In this proposal, we aim to characterize how EBV usurps B-cell maturation programs for cell survival and the establishment of a continuum of cell states balancing latency-driven proliferation, differentiation, and lytic reactivation. It is our central hypothesis that EBV establishes B-cell latent infection through mimicry of the germinal center (GC) reaction and subsequently promotes a continuum of activation and differentiation that is balanced to enable access to a cell state supporting lytic reactivation. We have formulated our central hypothesis based on preliminary data including single-cell gene expression, chromatin conformation of tonsillar B cells and EBV-immortalized cells as well as characterization of new spontaneously lytic strains of EBV. We found that EBV latent infection promotes GC mimicry through temporal regulation of anti-apoptotic MCL-1 and BFL-1. Using scRNA-seq of LCLs, we discovered a continuum of gene expression where NFB signaling/activation (e.g. NFKB2, MYC, IRF8) is strongly anti-correlated with plasmablast differentiation (e.g. CD38, PRDM1, XBP1). Furthermore, EBV lytic genes were expressed in cells most similar to the high differentiation state suggesting a model whereby EBV-infected cells constantly sample this differentiated state to enable a switch to lytic reactivation. Finally, using new strains of EBV we describe a paradigm of a persistent, spontaneous switch to productive infection that is transient and reversible. Therefore, the rationale for this proposed research is that understanding how EBV establishes latency and reactivates to productive infection provides insight into therapeutic modalities to eliminate EBV-infected cells from the oral cavity. The underlying molecular circuitry controlling these cell fate decisions may also provide important new information regarding the plasticity of B-cell maturation states. We plan to test our central hypothesis and complete the objectives in this proposal through the following three specific aims: i) to determine the molecular mechanisms by which EBV promotes B-cell survival mimicking tonsillar B-cell maturation, ii) to define the underlying molecular circuitry supporting a novel activation/differentiation continuum within individual EBV-immortalized B cells, and iii) to define the biochemical and cell biological features of a newly described EBV recurrence phenotype in which latently infected cells produce virions and return to their basal latent state.

抽象的我们的最终目标是定义口腔中 EBV 潜伏期建立和重新激活的分子机制在本提案中，我们旨在描述 EBV 如何篡夺 B 细胞成熟程序以促进细胞存活和发育。建立平衡潜伏驱动的增殖、分化和裂解的细胞状态连续体我们的中心假设是 EBV 通过模仿 B 细胞建立潜伏感染。生发中心 (GC) 反应，随后促进连续的激活和分化，即平衡以实现支持裂解重新激活的细胞状态。基于初步数据，包括单细胞基因表达、扁桃体 B 细胞染色质构象和 EBV 永生化细胞以及新的 EBV 自发裂解株的表征我们发现。 EBV 潜伏感染通过抗凋亡 MCL-1 和 BFL-1 的时间调节促进 GC 拟态。 LCL 的 scRNA-seq，我们发现了基因表达的连续体，其中 NF+B 信号/激活（例如 NFKB2、MYC、IRF8）与浆母细胞分化（例如 CD38、PRDM1、XBP1）强烈反相关。此外，EBV 裂解基因在与高分化状态最相似的细胞中表达，表明 EBV 感染细胞不断采样这种分化状态以实现裂解的模型最后，使用新的 EBV 毒株，我们描述了一种持续、自发转换的范例。生产性感染是短暂且可逆的，因此，这项研究的基本原理是：了解 EBV 如何建立潜伏期并重新激活产生性感染，可以深入了解从口腔中消除 EBV 感染细胞的治疗方式潜在的分子回路。控制这些细胞命运决定也可能提供有关 B 细胞可塑性的重要新信息我们计划通过以下方式测试我们的中心假设并完成本提案中的目标。以下三个具体目标： i) 确定 EBV 促进 B 细胞的分子机制模仿扁桃体 B 细胞成熟的生存，ii) 定义支持新颖的潜在分子电路个体 EBV 永生化 B 细胞内的激活/分化连续体，以及 iii) 定义生化以及新描述的 EBV 复发表型的细胞生物学特征，其中潜伏感染的细胞产生病毒体并返回其基础潜伏状态。