Targeting Epstein-Barr Virus Super-Enhancer

靶向 Epstein-Barr 病毒超级增强子

基本信息

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
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/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

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.

爱泼斯坦 - 巴尔病毒（EBV）引起感染性单核细胞增多症，淋巴瘤和淋巴增生性艾滋病毒感染和免疫抑制人的疾病与自身免疫性疾病有关。 EBV将静止的B淋巴细胞（RBL）转换为连续增殖的淋巴细胞细胞通过表达EBV核抗原（ENBAS）和潜在膜蛋白1（LMP1）的线（LCL）（LCLS）激活NF-KB。由于LCLS表达与某些EBV癌的同一EBV蛋白因此研究EBV在增长转化中的作用。 LCL的生长取决于EBNA2，EBNALP， EBNA3A，EBNA3C和LMP1。最近，我们发现所有必需的EBNA和LMP1 活化的NF-KB亚基汇聚为具有非凡的EBV超级增强剂（ESE） H3K27AC信号。 ESE控制着驱动LCL生长的关键致癌基因的表达，并且是对平均增强子的扰动更敏感。进一步表征分子组成及其在ESE中的功能作用，我们将（1）通过探测到其靶基因的循环，（2）确定ESE蛋白质组学组成并识别确定ESE形成的蛋白质，（3）确定ESE增强剂的功能作用 RNA（Erna）。我们将使用群集定期间隔短的短粒子重复序列（CRISPR）基于基于ESE循环至其直接靶基因必不可少的DNA元素的测定法和蛋白质对ESE活性必不可少的。我们将测试Erna敲击主机的效果转录和循环因子DNA结合。这里的实验使用了整合方法为了阐明分子机制激活关键的致癌潜水员。这些研究将确定治疗干预的机会。