Targeting Epstein-Barr Virus Super-Enhancer

靶向 Epstein-Barr 病毒超级增强子

• 批准号: 10379876
• 负责人: Bo Zhao
• 金额: $ 44.75万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10379876
• 关键词: AIDS-Related Lymphoma; Affect; Autoimmune Diseases; B-Lymphocytes; Binding; Binding Sites; Biological Assay; Biological Models; Bromodomain; CRISPR screen; CSPG6 gene; Carcinoma; Cell Line; Cells; ChIP-seq; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; DNA Binding; DNA Methylation; Development; Disease; EBV-associated disease; Elements; Enhancers; Epstein-Barr Virus Infections; Epstein-Barr Virus Nuclear Antigens; Epstein-Barr Virus-Related Malignant Neoplasm; Epstein-Barr pathogenesis; Gene Expression; Gene Proteins; Genes; Genetic Transcription; Growth; HIV; Health; Human; Human Herpesvirus 4; Immune; Immunoprecipitation; In Vitro; Infectious Mononucleosis; Intervention; Knock-out; Link; Lymphoma; Lymphoproliferative Disorders; Malignant Neoplasms; Membrane Proteins; Modeling; Molecular; NF-kappa B; Oncogenes; Oncogenic; Oncoproteins; Persons; Proliferating; Proteins; Proteomics; Quantitative Reverse Transcriptase PCR; RNA; RUNX3 gene; Reporter; Rest; Rheumatoid Arthritis; Role; SPI1 gene; Signal Transduction; Site; Testing; Therapeutic; Therapeutic Intervention; Viral Genes; Viral Proteins; base; cohesin; deep sequencing; experimental study; genetic manipulation; genome-wide; inhibitor; insight; knock-down; lymphoblast; lymphoblastoid cell line; novel therapeutics; post-transplant; programs; small hairpin RNA; transcription factor; tumorigenesis; virus related cancer; young adult

Epstein-Barr Virus (EBV) causes infectious mononucleosis, lymphomas and lymphoproliferative diseases in HIV infected and immune suppressed people and is linked to autoimmune diseases. EBV converts Resting B Lymphocytes (RBLs) to continuously proliferating Lymphoblasts Cell Lines (LCLs) by expressing EBV nuclear antigens (ENBAs) and latent membrane protein 1 (LMP1) that activates NF-kB. Since LCLs express that same EBV proteins as some EBV cancers, EBV conversion of RBLs to LCLs is therefore a relevant model that can be genetically manipulated to investigate EBV's role in growth transformation. LCL growth depends on EBNA2, EBNALP, EBNA3A, EBNA3C and LMP1. Recently, we found that all the essential EBNAs and LMP1 activated NF-kB subunits converge to EBV super-enhancers (ESE) that have extraordinary H3K27ac signals. ESEs govern the expression of key oncogenes that drive LCL growth and are more sensitive to perturbations that average enhancers. To further characterize the molecular composition and their functional roles in ESEs, we will (1) Determine the mechanisms through which ESEs loop to their target genes, (2) Determine the ESE proteomic composition and identify proteins that determine ESE formation, and (3) Determine the functional roles of ESE enhancer RNA (eRNA). We will use Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) based assays to identify the DNA elements essential for ESE to loop to their direct target genes and proteins essential for ESE activity. We will test the effect of eRNA knock down on host transcription and looping factor DNA binding. The experiments here in use integrative approaches to elucidate the molecular mechanism by which ESEs activate key oncogenic divers. These studies will identify opportunities for therapeutic intervention.

EB病毒（EBV）引起传染性单核细胞增多症、淋巴瘤和淋巴增生性淋巴瘤。 艾滋病毒感染者和免疫抑制者的疾病，并与自身免疫性疾病有关。 EBV将静息B淋巴细胞（RBL）转化为持续增殖的淋巴母细胞 通过表达EBV核抗原（ENBA）和潜伏膜蛋白1（LMP 1） 能激活NF-κ B由于LCL表达与某些EBV癌症相同的EBV蛋白， 因此，将RBL转化为LCL是一种相关的模型，可以通过遗传操作， 研究EBV在生长转化中的作用。LCL增长取决于EBNA 2，EBNALP， EBNA 3A、EBNA 3C和LMP 1。最近，我们发现所有必需的EBNA和LMP 1 激活的NF-κ B亚单位会聚到EBV超级增强子（ESE）， H3 K27 ac信号。ESE控制着驱动LCL生长的关键癌基因的表达， 对平均增强子的扰动更敏感。为了进一步表征 组成和他们的职能作用，在环境和社会经济，我们将（1）确定机制，通过 （2）确定ESE蛋白质组组成，并鉴定 决定ESE形成的蛋白质，和（3）确定ESE增强子的功能作用 RNA（eRNA）。我们将使用重复的规则间隔短回文重复序列（CRISPR） 用于鉴定ESE与其直接靶基因形成环所必需的DNA元件 和ESE活性所必需的蛋白质。我们将测试eRNA敲低对宿主的影响， 转录和成环因子DNA结合。这里的实验采用综合方法 阐明ESE激活关键致癌因子的分子机制。这些 研究将确定治疗干预的机会。