EPSTEIN BARR VIRUS LMP1 MEDIATED ONCOGENICITY

Epstein Barr 病毒 LMP1 介导的致癌性

• 批准号: 6776477
• 负责人: ELLIOTT D KIEFF
• 金额: $ 72.63万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-20 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6776477
• 关键词: AP1 protein; B lymphocyte; CD40 molecule; Caenorhabditis elegans; Epstein Barr virus; I kappa B beta; apoptosis; cAMP response element binding protein; cell membrane; clinical research; cytokine receptors; cytoskeleton; epidermal growth factor; genetic promoter element; genetic transcription; genetically modified animals; growth factor receptors; human subject; laboratory mouse; membrane proteins; neoplasm /cancer genetics; nuclear factor kappa beta; recombinant virus; tumor necrosis factor alpha; viral carcinogenesis; virus genetics; virus protein

DESCRIPTION: (as per abstract) The objective is to develop sufficient knowledge of the molecular mechanisms by which LMP1 alters cell growth and of the effects of inhibiting those mechanisms so as to design screens for inhibitors that might be effective in treating EBV associated malignancies. The specific goals are to: (1) Use recombinant EBV based genetics to clarify the role of specific LMP1 amino acids (aa) in primary B lymphocyte growth transformation. The focus will be on aa between transformation effector site 1 (TES1) and TES2, TES1, TES2 and aa within the transmembrane domains. (2) Determine whether LMP1 signals through interactions with other receptors. LMP1 association with membrane rafts, with cytoskeletal elements, and with other membrane proteins, particularly TNFRs, will be investigated using genetic, biochemical, and physiological approaches. (3) Determine the mechanisms by which N-terminally truncated IkB causes apoptosis of EBV transformed cells so as to better validate inhibition of NFkB as a therapeutic target. (4) Investigate the cytoplasmic effects of LMP1, including the molecular mechanisms by which LMP1 and CD40 recruit TRAFs, modify TRAFs, activate NIK and ASK, effect NIK and ASK1 interaction with downstream kinases, and continuously signal or return to a basal state. (5) Determine the effects of LMP1 on B lymphocytes and epithelial gene expression from the changes in RNAs following expression of LMP1, LMP1 TES1, LMP1 TES2, or LMP1 with null mutations in TES1 and TES2. The importance of NFkB, AP1, CREB, or other factor activations will be evaluated by analyses of relevant promoters, e.g., TRAF1, EGFR, and CD40. The effects of LMP1TES1 and LMP1TES2 on cell gene transcription and on transformation will be compared with the effects of LMP1 and LMP1/CD40 in transgenic mice. (6) Evaluate the role of TRAF3 in LMP1, CD40, and LTbetaR signaling through observing differences in RNAs with LMP1 expression or CD40 or LTbetaR activation in TRAF3 knockout versus normal mouse cells or mice. The role of PKN in TRAF3 effects on the cell cytoskeleton, membranes, or transcription will also be assessed. (7) Investigate the role of C. elegans TRAF3 homologue in C. elegans neural function so as to enable screens for interacting genes that might identify novel TRAF functions.

描述：（根据摘要）目标是发展足够的知识 LMP1 改变细胞生长的分子机制及其影响 抑制这些机制，以便设计抑制剂的筛选 可能有效治疗 EBV 相关恶性肿瘤。具体目标 目的是： (1) 使用基于重组 EBV 的遗传学来阐明特定的作用 LMP1 氨基酸 (aa) 在原代 B 淋巴细胞生长转化中的作用。焦点 将位于转化效应位点 1 (TES1) 和 TES2、TES1 之间的氨基酸上， TES2 和 aa 位于跨膜域内。 (2)判断LMP1是否 通过与其他受体相互作用发出信号。 LMP1 与 膜筏，具有细胞骨架元件和其他膜蛋白， 特别是 TNFR，将通过遗传、生化和 生理学方法。 (3) 确定N端的机制 截短的IkB导致EBV转化细胞凋亡，从而更好地 验证 NFkB 抑制作为治疗靶点。 (4) 调查 LMP1 的细胞质效应，包括 LMP1 的分子机制 CD40招募TRAF，修改TRAF，激活NIK和ASK，影响NIK和ASK1 与下游激酶相互作用，并持续发出信号或返回到 基础状态。 (5)确定LMP1对B淋巴细胞和上皮细胞的影响 LMP1、LMP1 表达后 RNA 变化的基因表达 TES1、LMP1 TES2 或 TES1 和 TES2 中具有无效突变的 LMP1。重要性 NFkB、AP1、CREB ​​或其他因子的激活将通过分析进行评估 相关启动子，例如 TRAF1、EGFR 和 CD40。 LMP1TES1 和的影响 LMP1TES2 对细胞基因转录和转化的影响将与 LMP1 和 LMP1/CD40 对转基因小鼠的影响。 (6) 评估角色 通过观察 LMP1、CD40 和 LTbetaR 信号转导中的 TRAF3 差异 TRAF3 敲除中具有 LMP1 表达或 CD40 或 LTbetaR 激活的 RNA 与正常小鼠细胞或小鼠相比。 TRAF3中PKN对细胞的作用 细胞骨架、膜或转录也将被评估。 (7) 研究秀丽隐杆线虫 TRAF3 同源物在秀丽隐杆线虫神经中的作用 功能，以便能够筛选可能识别的相互作用基因 新颖的 TRAF 功能。