Regulation of Latent-Lytic Switch in EBV by ZEB

描述（由申请人提供）：EB病毒（EBV）是一种人γ疱疹病毒，在成年期感染大多数人。感染与传染性单核细胞增多症、伯基特和非霍奇金淋巴瘤、霍奇金病、鼻咽癌、口腔毛状白斑和某些胃癌有关。这项研究的长期目标是（i）阐明控制潜伏期与EBV裂解性复制的机制，以及（ii）将这些知识应用于治疗某些EBV相关疾病的新疗法的开发。Mertz博士和她的同事最近发现了一种新的顺式作用元件ZV及其反式作用因子ZEB 1，它们在调节EBV潜伏裂解开关基因BZLF 1的潜伏期及其通过TGF-β和抗免疫球蛋白等诱导剂的重新激活中发挥核心作用。具体目标如下：（i）确定ZEB 1在通过结合BZLF 1启动子中的ZV元件调节EBV生命周期中的重要性，通过（a）确定ZV元件中的突变对建立潜伏期和裂解性复制的再激活的影响，（B）证实ZEB 1通过结合ZV元件调节BZLF 1启动子的表达，（c）使用可诱导表达ZEB 1的细胞系确定ZEB 1的丰度对潜伏期和再激活的建立和维持的影响，和（d）通过测量多种人B-细胞中ZEB 1 mRNA和蛋白质的丰度，寻找ZEB 1的存在与EBV感染后果之间的相关性。淋巴细胞和上皮正常和EBV相关肿瘤组织;和（ii）阐明细胞信号传导途径的效应物部分通过调节ZEB 1诱导EBV潜伏或再活化的机制，通过（a）使用可诱导表达EBV的LMP 1和LMP 1突变体的细胞，测定EBV编码的蛋白质LMP 1对ZEB 1丰度、活性的影响和对EBV生命周期的影响，和B）通过寻找表达与用抗免疫球蛋白和TGF-β激活细胞后ZEB 1的丰度、细胞定位、DNA结合活性、共纯化共调节因子和翻译后修饰的变化之间的相关性，测试通过ZEB 1激活BZLF 1基因表达的模型。这些研究证实了我们关于ZEB 1和ZV元件在调节EBV生命周期中的核心作用的假设，并可能导致针对EBV的免疫和一些EBV相关疾病的治疗的新疗法。

DESCRIPTION (provided by applicant): Epstein-Barr virus (EBV) is a human gamma herpesvirus that infects most people by adulthood. Infection is associated with infectious mononucleosis, Burkitt's and non-Hodgkin's lymphomas, Hodgkin's disease, nasopharyngeal carcinoma, oral hairy leukoplakia, and some gastric carcinomas. The long-term goals of this research are (i) to elucidate mechanisms controlling latency versus lytic replication of EBV, and (ii) to apply this knowledge toward the development of new therapies for treating some EBV-associated diseases. Dr. Mertz and her colleagues have recently identified a novel cis-acting element, ZV, and its trans-acting factor, ZEB1, that play central roles in regulating EBV's latent-lytic switch gene, BZLF1, during latency and its reactivation by inducers such as TGF-beta and anti-immunoglobulins. The specific aims are the following: (i) Determine importance of ZEB1 in regulating EBV's life cycle via binding the ZV element in the BZLF1 promoter by (a) determining effects of mutations in the ZV element on establishment of latency and reactivation to lytic replication, (b) confirming ZEB1 regulates expression of the BZLF1 promoter via binding the ZV element, (c) determining the effects of ZEB1's abundance on establishment and maintenance of latency and reactivation using cell lines inducible for expression of ZEB1, and (d) looking for correlations between presence of ZEB1 and consequences of infection by EBV by measuring the abundance of ZEB1 mRNA and protein in a variety of human B-lymphocytic and epithelial normal and EBV-associated tumor tissues; and (ii) Elucidate mechanisms by which effectors of cellular signaling pathways induce latency or reactivation of EBV in part via modulating ZEB1 by (a) determining the effects of the EBV-encoded protein LMP1 on ZEB1's abundance, activity, and effects on EBV's life cycle using cells inducible for expression of LMP1 and LMP1 mutants of EBV, and b) testing models for activation of expression of the BZLF1 gene via ZEB1 by looking for correlations between expression and changes in the abundance, cellular localization, DNA-binding activity, co-purifying co-regulators, and post-translational modifications of ZEB1 after activation of cells with anti-immunoglobulins and TGF-beta. These studies should confirm our hypothesis regarding the central roles of ZEB1 and the ZV element in regulating EBV's life cycle and, possibly, lead to novel therapies for immunization against EBV and treatment of some EBV-associated diseases.