Epstein-Barr Virus LMP1-Mediated Oncogenecity

描述（由申请人提供）：爱泼斯坦-巴尔病毒是霍奇金病、淋巴瘤和恶性增殖性疾病的重要病因，特别是在HIV感染和其他免疫受损状态的人群中。EBV癌蛋白潜伏膜蛋白1通过B细胞生长的2个必需信号传导结构域转化B细胞：转化效应位点1，其激活非典型NF κ B;和位点2，其激活典型NF κ B。两种NFkB途径对于感染细胞的生长和存活都是必需的。为了发现LMP 1影响NFkB通路的关键组分，我们的目标是：（1）表征LMP 1 TES 2典型NFkB活化所需的关键B细胞蛋白，以确定它们在TRAF 6和IKK活化、NEMO泛素化和核RelA磷酸化中的作用。缺失激酶、磷酸酶、E3连接酶和支架的候选物已经通过293细胞中的全基因组siRNA筛选鉴定，并将在B细胞中进行评价。(2)将鉴定LMP 1 TES 1非典型NFkB活化所需的关键B细胞蛋白。这一目标将集中在独特的LMP 1通过TRAF的影响，将采用LMP 1和TRAF遗传分析，并将确定新的细胞蛋白的关键LMP 1介导的非经典NF κ B激活。(3)识别 AIM 1和2靶蛋白敲低的组合，当两者都从EBV转化的B细胞中耗尽时，其产生合成的致死效应。AIM 3实验利用了我们在AIMS 1和2中所学到的知识来识别EBV相关淋巴瘤的阿基里斯之踵。 对LMP 1介导的NFkB活化不具有特异性的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应答。这些研究通过确定EBV影响肿瘤促进和进展的机制，通过发现用于治疗患有AIDS定义的恶性肿瘤的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药物，专门解决PA 10 -290中概述的NCI目标。我们的发现将阐明NFkB激活的新关键靶点，并更广泛地推进基于靶点的优先工具化合物发现。

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.