Epstein-Barr Virus LMP-1 Mediated Oncogenicity

描述(申请人提供)：这项研究计划集中在Epstein-Barr病毒潜伏膜蛋白1(LMP1)信号转导，特别是识别对LMP1介导的NF-kB激活至关重要的分子靶标，并通过遗传和化学方法验证靶标。具体目标1是利用反向遗传、生化和荧光共振能量转移方法表征LMP1跨膜相互作用，这些作用对LMP1介导的NF-kB激活至关重要。在这些研究中获得的知识可能使跨膜聚集和信号转导中断。具体目标2通过串联亲和下拉实验鉴定TRAF2、TRAF3、NIK、Tradd、TRAF6和IRAKI相关蛋白，从而识别LMP1 TES1和TES2 NF-kB激活的缺失成分。可能的信号成分将通过在B淋巴细胞和上皮细胞中使用siRNA技术定向敲除来评估它们在NF-kB激活中的作用。基于siRNA的遗传筛选也将用于识别核因子-kB途径的基本成分。具体目标3是基于细胞的筛选LMP1诱导的核因子-KB的新型化学抑制剂，可作为新疗法的化学核心。还将确定这些工具化合物的目标。核因子-kB活性是EBV转化的B淋巴细胞存活所必需的。因此，LMP1核因子-kB激活的抑制剂将有助于治疗LMP1表达的EBV相关疾病，包括艾滋病患者和移植受者的淋巴增生性疾病、霍奇金氏病和鼻咽癌。鉴于核因子-kB活性在淋巴细胞激活、生长、分化和存活中的重要性，工具化合物也可能为适用于过敏、移植或自身免疫性疾病的药物开发提供方向。

DESCRIPTION (provided by applicant): This research program focuses on Epstein-Barr virus Latent membrane protein 1 (LMP1) signaling, particularly the identification of molecular targets critical for LMP1-mediated NF-kB activation and target validation through genetic and chemical approaches. Specific objective 1 is to characterize LMP1 transmembrane interactions that are critical for LMP1 mediated NF-kB activation using reverse genetic, biochemical and fluorescence resonance energy transfer methodologies. Knowledge gained in these studies may enable interruption of transmembrane aggregation and signaling. Specific objective 2 focuses on identifying missing components of LMP1 TES1 and TES2 NF-kB activation by characterizing TRAF2, TRAF3, NIK, TRADD, TRAF6 and IRAKI associated proteins by tandem affinity pull-down experiments. Putative signaling components will be evaluated for their role in NF-kB activation by directed knock out using siRNA technology in B lymphocytes and epithelial cells. An siRNA based genetic screen will also be used to identify essential components of the NF-kB pathway. Specific objective 3 is a cell based screen for novel chemical inhibitors of LMP1 induced NF-KB that can serve as a chemical nucleus for novel therapeutics. The targets of such tool compounds will also be determined. NF- kB activity is required for EBV transformed B-lymphocyte survival. Thus inhibitors of LMP1 NF- kB activation will be useful for treating EBV associated diseases where LMP1 is expressed including lymphoproliferative disease in AIDS patients and transplant recipients, Hodgkin's disease, and Nasopharyngeal carcinoma. Given the importance of NF-kB activity in lymphocyte activation, growth, differentiation and survival, a tool compound might also provide direction for drug discovery applicable for allergy, transplantation, or auto immune disease.