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.

抽象的我们定义EBV潜伏期建立和重新激活的分子机制的最终目标缓存。在此提案中，我们旨在表征EBV如何篡夺B细胞成熟计划，以用于细胞生存和建立一个细胞状态的连续体平衡潜伏期驱动的增殖，分化和裂解重新激活。我们的核心假设是，EBV通过模仿模仿建立B细胞潜在感染生发中心（GC）反应，随后促进了激活和分化的连续性平衡以使能够访问支持裂解重新激活的细胞状态。我们已经提出了中心假设基于初步数据，包括单细胞基因表达，扁桃体B细胞的染色质构象和 EBV侵蚀性细胞以及EBV的新赞助商裂解菌株的表征。我们发现 EBV潜在感染通过抗凋亡MCL-1和BFL-1的临时调节促进GC模拟。使用 LCLS的SCRNA-SEQ，我们发现了基因表达的连续体，其中NFB信号传导/激活（例如， NFKB2，MYC，IRF8）与浆膜分化非常抗相关（例如CD38，PRDM1，XBP1）。此外，EBV裂解基因在与高分化状态最相似的细胞中表达模型，通过该模型，EBV感染的单元不断采样了这种差异化状态以使转换为裂解重新激活。最后，使用新的EBV菌株，我们描述了一个持续的，赞的范式瞬时和可逆的生产感染。因此，这项拟议研究的理由是了解EBV如何建立潜伏期并重新激活产品感染从口腔中消除EBV感染的细胞的治疗方式。基础分子电路控制这些细胞脂肪决策也可能提供有关B细胞可塑性的重要新信息成熟状态。我们计划通过以下三个特定目的：i）确定EBV促进B细胞的分子机制生存模仿扁桃体B细胞的成熟，ii）定义支持新颖的基本分子电路激活/分化在单个EBV侵蚀B细胞中继续进行，并在iii中定义生化以及新描述的EBV复发表型的细胞生物学特征，其中潜在感染细胞产生病毒并返回其基本潜在状态。