Regulation of the Epstein-Barr Virus Lytic Switch

Epstein-Barr 病毒裂解开关的调节

基本信息

批准号：
10317642
负责人：
Benjamin Elison Gewurz
金额：
$ 50.75万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-06 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10317642
关键词：
3-Dimensional Adult Antigens Architecture Auxins B-Lymphocytes BZLF1 gene Binding Binding Sites CRISPR screen Cell Compartmentation Cell Nucleus Cells ChIP-seq Chromatin Conformation Capture and Sequencing Chromatin Loop Clustered Regularly Interspaced Short Palindromic Repeats Complex Cues DNA Polymerase II EBV-associated disease Engineering Epigenetic Process Epithelial Epstein-Barr Virus latency Gene Expression Gene Expression Regulation Genetic Transcription Genome Hairy Leukoplakia Human Genome Human Herpesvirus 4 Immediate-Early Genes Immune response Immunoglobulins Infection Infectious Mononucleosis Integration Host Factors Knowledge Lead Location Lymphoma Lytic Lytic Phase Lytic Virus Malignant Neoplasms Malignant neoplasm of nasopharynx Maps Mediator of activation protein Memory B-Lymphocyte Methylation Molecular Conformation Nasopharynx Carcinoma Oral cavity PRDM1 gene Pathway interactions Phosphorylation Physiological Proto-Oncogene Proteins c-myc RNA Regulation Resolution Role Saliva Series Stomach Carcinoma Structure System Therapeutic Tonsil Transcription Factor AP-1 Transcriptional Regulation Viral Viral Genome Viral Proteins Virion Withdrawal Xenograft procedure cohesin crosslink ds-DNA genome-wide in vivo insight lytic replication malignant stomach neoplasm novel therapeutic intervention plasma cell differentiation programs promoter release factor response single molecule transcription factor tumorigenesis viral genomics

项目摘要

Abstract Epstein-Barr virus (EBV) establishes lifelong infection in 95% of adults worldwide. EBV causes infectious mononucleosis, lymphomas, nasopharyngeal and gastric cancer, and oral hairy leukoplakia. EBV is transmitted between hosts through saliva, from which it translocates across the oral cavity and tonsillar epithelium to reach the B-cell compartment. Upon B-cell infection, the ~170 kilobase linear double-stranded DNA (dsDNA) EBV genome is delivered to the nucleus. EBV then uses a series of latency programs to navigate the B-cell compartment, and colonize the memory B-cell compartment. Ultimately, to spread between cells and to the tonsillar epithelium for transfer to a new host, EBV must undergo lytic replication, in which nearly 80 viral proteins are expressed and infectious virion are produced. Lytic replication is increasingly implicated in oncogenesis. Yet, much remains to be learned about how the viral lytic switch is controlled. We therefore used a human genome-wide CRISPR/Cas9 screen to identify host factors that control the viral lytic switch. Our analyses a network of host factors that repress lytic reactivation, centered on the transcription factor MYC, including cohesins, FACT, STAGA, and Mediator. Depletion of MYC or factors important for MYC expression, reactivated the lytic cycle, including in Burkitt xenografts. MYC bound the EBV genome origin of lytic replication and suppressed its looping to the lytic cycle initiator BZLF1 promoter. Our central hypothesis is that the MYC:MAX control the EBV lytic switch through regulation of viral genome loop extrusion. Our Aims are to (1) Define key MYC roles in regulation of oriLyt loop extrusion to control the EBV lytic switch; (2) Define MYC-gated cohesin roles in regulation of oriLyt loop extrusion in EBV lytic switch control (3) Define MYC-gated CTCF roles in regulation of oriLyt loop extrusion to control the EBV lytic switch. Collectively, these studies are expected to identify how EBV subverts host DNA loop extrusion pathways to control the viral lytic switch. Our studies may therefore support strategies to develop rational lytic induction therapeutic therapies for EBV-associated diseases.

抽象的爱泼斯坦-巴尔病毒 (EBV) 导致全球 95% 的成年人终身感染。 EB病毒引起传染性单核细胞增多症、淋巴瘤、鼻咽癌、胃癌以及口腔癌毛状白斑。 EBV 通过唾液在宿主之间传播，并从唾液中转移穿过口腔和扁桃体上皮到达 B 细胞室。 B细胞上感染后，约 170 KB 的线性双链 DNA (dsDNA) EBV 基因组被传递到核。然后 EBV 使用一系列延迟程序来导航 B 细胞室，并定殖于记忆 B 细胞区室。最终，在细胞之间传播并传播到为了转移到新宿主，EBV 必须经历扁桃体上皮的裂解复制，其中近 80 种病毒蛋白被表达并产生感染性病毒颗粒。裂解复制是越来越多地参与肿瘤发生。然而，关于病毒如何传播，还有很多东西有待了解。控制溶解开关。因此，我们使用人类全基因组 CRISPR/Cas9 筛选来确定控制病毒裂解开关的宿主因素。我们分析了一系列宿主因素抑制裂解再激活，以转录因子 MYC 为中心，包括粘连蛋白、FACT、 STAGA 和调解员。 MYC 或对 MYC 表达重要的因子耗尽，重新激活裂解周期，包括伯基特异种移植物。 MYC结合EBV基因组裂解起源复制并抑制其与裂解循环启动子 BZLF1 启动子的循环。我们的中央假设 MYC:MAX 通过调节病毒基因组来控制 EBV 裂解开关循环挤出。我们的目标是 (1) 定义 MYC 在 oriLyt 环挤出调节中的关键作用控制 EBV 裂解开关； (2) 定义 MYC 门控粘连蛋白在 oriLyt 环调节中的作用 EBV 裂解开关控制中的挤出 (3) 定义 MYC 门控 CTCF 在 oriLyt 调节中的作用循环挤出来控制 EBV 裂解开关。总的来说，这些研究预计将确定 EBV 如何破坏宿主 DNA 环挤出途径来控制病毒裂解开关。因此，我们的研究可能支持开发合理的裂解诱导治疗的策略 EBV 相关疾病的治疗。