Targeting Epstein-Barr Virus Super-Enhancer

EB病毒可引起传染性单核细胞增多症、淋巴瘤和淋巴增生性疾病 艾滋病毒感染者的疾病和免疫抑制，并与自身免疫性疾病有关。 EB病毒将静息B淋巴细胞转化为持续增殖的淋巴母细胞 表达EB病毒核抗原和潜伏膜蛋白1的细胞系(LCL) 从而激活核因子-kB。由于LCL表达与某些EBV癌相同EBV蛋白，因此EBV 因此，RBL到LCLS的转换是一个相关的模型，可以通过基因操作来实现 研究EBV在增长转型中的作用。LCL的增长依赖于EBNA2、EBNALP、 EBNA3A、EBNA3C和LMP1。最近，我们发现所有必需的EBNAs和LMP1 激活的核因子-kB亚基会聚为EBV超级增强子(ESE)，具有非凡的 H3K27ac信号。ESES控制推动LCL生长的关键癌基因的表达，并 对平均增强剂的扰动更加敏感。为了进一步确定分子的特征 ESES的组成及其功能作用，我们将(1)通过以下方式确定其机制 (2)确定ESE的蛋白质组组成并鉴定 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)。我们将使用聚簇规则间隔短回文重复(CRISPR) 基于鉴定ESE与其直接靶基因环路所必需的DNA元件的分析 和ESE活性所必需的蛋白质。我们将测试Erna击倒对主机的影响 转录和环状因子DNA结合。这里的实验使用的是综合方法 阐明ESES激活关键致癌潜水器的分子机制。这些 研究将确定治疗干预的机会。

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.