Regulation of Latent-Lytic Switch in EBV by ZEB

描述(申请人提供)：爱泼斯坦-巴尔病毒(EBV)是一种人类伽玛疱疹病毒，成年后感染大多数人。感染与传染性单核细胞增多症、伯基特氏和非霍奇金淋巴瘤、霍奇金氏病、鼻咽癌、口腔毛状白斑和一些胃癌有关。这项研究的长期目标是(I)阐明EBV潜伏期与裂解复制的控制机制，以及(Ii)将这一知识应用于开发治疗某些EBV相关疾病的新疗法。Mertz博士和她的同事们最近发现了一种新的顺式作用元件ZV及其反式作用因子ZEB1，它们在潜伏期调控EBV的潜伏裂解开关基因BZLF1，并通过转化生长因子-β和抗免疫球蛋白等诱导剂重新激活它，发挥着核心作用。具体目的如下：(I)通过与BZLF1启动子中的ZV元件结合来确定ZEB1在调节EB病毒生命周期中的重要性，方法是(A)确定ZV元件突变对建立潜伏期和重新激活裂解复制的影响，(B)确认ZEB1通过结合ZV元件调节BZLF1启动子的表达，(C)利用可诱导ZEB1表达的细胞系确定ZEB1的S丰度对潜伏期和再激活的建立和维持的影响，以及(D)通过测量各种人类B淋巴细胞和上皮性正常和EBV相关肿瘤组织中ZEB1的mRNA和蛋白的丰度，寻找ZEB1的存在与EBV感染后果之间的相关性；以及(Ii)阐明细胞信号通路的效应者通过以下方式部分通过调节ZEB1来诱导EBVBZLF1潜伏或重新激活的机制：(A)利用可诱导表达EBVLMP1和EBVLMP1突变体的细胞，确定EBVLMP1对ZEB1‘S丰度、活性和对EBV生活史的影响；以及(B)通过寻找ZEB1在抗免疫球蛋白和转化生长因子-β激活细胞后的表达和变化之间的关系，来测试ZEB1激活BZLF1基因表达的模型。这些研究将证实我们关于ZEB1和ZV元件在调节EBV生命周期中的中心作用的假设，并可能导致针对EBV的免疫和某些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.