Targeting Epstein-Barr Virus Super-Enhancer

针对 Epstein-Barr 病毒超级增强子

基本信息

批准号：
9970995
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
项目类别：
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/9970995
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 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/antagonist insight knock-down lymphoblast lymphoblastoid cell line novel therapeutics post-transplant programs small hairpin RNA transcription factor tumorigenesis virus related cancer young adult

项目摘要

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) 导致传染性单核细胞增多症、淋巴瘤和淋巴增殖性疾病艾滋病毒感染者和免疫抑制人群的疾病，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细胞表达 EBV 核抗原 (ENBA) 和潜伏膜蛋白 1 (LMP1) 的细胞系 (LCL) 激活 NF-kB。由于 LCL 表达与某些 EBV 癌症相同的 EBV 蛋白，因此 EBV 因此，RBL 到 LCL 的转换是一个相关模型，可以通过基因操作来实现研究 EBV 在生长转化中的作用。拼箱增长取决于 EBNA2、EBNALP、 EBNA3A、EBNA3C 和 LMP1。最近，我们发现所有必需的 EBNA 和 LMP1 激活的 NF-kB 亚基会聚至 EBV 超级增强子 (ESE)，该增强子具有非凡的作用 H3K27ac 信号。 ESE 控制驱动 LCL 生长的关键癌基因的表达，并且比平均增强子对扰动更敏感。为了进一步表征分子组成及其在 ESE 中的功能作用，我们将 (1) 通过以下方式确定机制：哪些 ESE 循环至其靶基因，(2) 确定 ESE 蛋白质组组成并识别决定 ESE 形成的蛋白质，以及 (3) 确定 ESE 增强子的功能作用 RNA（eRNA）。我们将使用成簇规则间隔短回文重复序列 (CRISPR) 基于检测来识别 ESE 循环至其直接目标基因所必需的 DNA 元件以及 ESE 活性必需的蛋白质。我们将测试eRNA敲低对宿主的影响转录和环因子 DNA 结合。这里的实验使用综合方法阐明 ESE 激活关键致癌因素的分子机制。这些研究将确定治疗干预的机会。