Epstein-Barr Virus LMP-1 Mediated Oncongenicity

描述（由申请人提供）：该研究项目的重点是EB病毒潜伏膜蛋白1（LMP 1）信号传导，特别是通过遗传和化学方法鉴定对LMP 1介导的NF-κ B活化至关重要的分子靶标和靶标验证。具体目标1是使用反向遗传、生化和荧光共振能量转移方法来表征对LMP 1介导的NF-κ B活化至关重要的LMP 1跨膜相互作用。在这些研究中获得的知识可能能够中断跨膜聚集和信号传导。具体目标2侧重于通过串联亲和下拉实验表征TRAF 2、TRAF 3、NIK、TRADD、TRAF 6和IRAKI相关蛋白来鉴定LMP 1 TES 1和TES 2 NF-kB活化的缺失组分。通过在B淋巴细胞和上皮细胞中使用siRNA技术定向敲除，评价推定的信号传导组分在NF-κ B活化中的作用。基于siRNA的遗传筛选也将用于鉴定NF-kB途径的必要组分。具体目的3是基于细胞筛选LMP 1诱导的NF-κ B的新型化学抑制剂，其可用作新型治疗剂的化学核。还将确定这些工具化合物的目标。NF-kB活性是EBV转化的B淋巴细胞存活所必需的。因此，LMP 1 NF-kB活化的抑制剂可用于治疗EBV相关疾病，其中LMP 1被表达，包括AIDS患者和移植受体中的淋巴增生性疾病、霍奇金病和鼻咽癌.鉴于NF-kB活性在淋巴细胞活化、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、移植或自身免疫疾病的药物发现提供方向。

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.