Epstein-Barr Virus LMP1-Mediated Oncogenecity

描述(申请人提供)：爱泼斯坦-巴尔病毒是霍奇金氏病、淋巴瘤和淋巴增殖性疾病的重要病因，特别是在艾滋病毒感染和其他免疫受损状态的人中。EB病毒癌蛋白潜伏膜蛋白1通过B细胞生长所必需的两个信号域来转化B细胞：转化效应位点1，激活非典型性NFkB；和位点2，激活典型性NFkB。这两条NFkB通路对于感染细胞的生长和存活都是必要的。为了发现LMP1影响NFkB通路的关键成分，我们的目标是：(1)鉴定LMP1 TES2规范的NFkB激活所需的关键B细胞蛋白在TRAF6和IKK激活、Nemo泛素化和核RelA磷酸化中的作用。在293细胞中，通过全基因组siRNA筛选已经确定了缺失的激酶、磷酸酶、E3连接酶和支架的候选基因，并将在B细胞中进行评估。(2)将鉴定LMP1 TES1非规范NFkB激活所需的关键B细胞蛋白。这一目标将集中于通过TRAF的独特的LMP1效应，将使用LMP1和TRAF的遗传分析，并将识别对LMP1介导的非规范NFkB激活至关重要的新的细胞蛋白质。(3)识别 当两者从EBV转化的B细胞中被耗尽时，AIM 1和2靶蛋白的组合被击倒，从而产生合成的致死效应。AIM3实验利用我们在AIMS 1和2中学到的东西来识别EBV相关淋巴瘤的跟腱。对于LMP1介导的NFkB激活没有特异性的NFkB小分子抑制剂可能会受到副作用的限制，包括抑制关键的免疫反应。这些研究特别针对PA10-290中概述的NCI目标，通过确定EBV影响肿瘤促进和进展的机制，通过发现治疗艾滋病定义的恶性肿瘤患者的合理药物发现的新靶点，以及通过使用组合基因组方法进一步开发治疗剂。我们的发现将阐明NFkB激活的新的关键靶点，并更广泛地推进基于优先靶点的工具化合物的发现。

DESCRIPTION (provided by applicant): Epstein - Barr virus is an important cause Hodgkin's Disease, Lymphomas, and Lymphoproliferative Diseases, particularly in people with HIV infection and other immune- compromised states. The EBV oncoprotein Latent Membrane Protein 1 transforms B-cells through 2 essential signaling domains for B cell growth: Transformation effect site 1, which activates non- canonical NFkB and site 2, which activates canonical NFkB. Both NFkB pathways are necessary for infected cell growth and survival. To discover key components of LMP1 affected NFkB pathways, our AIMS are to: (1) Characterize key B-cell proteins required for LMP1 TES2 canonical NFkB activation for their role in TRAF6 and IKK activation, NEMO ubiquitination, and nuclear RelA phosphorylation. Candidates for missing kinases, phosphatases, E3 ligases, and scaffolds have been identified through a genome wide siRNA screen in 293 cells and will be evaluated in B cells. (2) Key B-cell proteins required for LMP1 TES1 non-canonical NFkB activation will be identified. This aim will focus on unique LMP1 effects through TRAFs, will employ LMP1 and TRAF genetic analyses and will identify novel cell proteins critical for LMP1-mediated non-canonical NFkB activation. (3) Identify combinations of AIM 1 and 2 target proteins knockdowns that create synthetic lethal effects, when both are depleted from EBV-transformed B cells. AIM3 experiments exploit what we learn in AIMS 1 and 2 to identify the Achilles' Heel of EBV-associated lymphomas. NFkB small molecule inhibitors that are not specific for LMP1-mediated NFkB activation will likely be limited by side-effects, including inhibition of critical immune responses. These studies specifically address NCI goals outlined in PA10-290 by determining the mechanism by which EBV affects tumor promotion and progression, by discovering novel targets for rational drug discovery for the treatment of persons afflicted with AIDS-defining malignancies, and by using combinatorial genomic methodologies to further development of therapeutic agents. Our discoveries will elucidate new key targets in NFkB activation and more broadly advance priority target-based tool compound discovery.