Plasmid Replicons of Human Tumor Viruses

我们想了解肿瘤前病变和肿瘤后病变中人类肿瘤病毒的独特功能， 肿瘤细胞这些功能是特异性抗病毒治疗的潜在靶点，以治疗病毒相关性疾病。 癌的我们（Lambert和Sugden博士）已经研究了EBV和HPV的复制， 质粒复制子是已知的，并且最近将我们的工作扩展到包括KSHV。我们的目标 研究已经详细描述了这些病毒复制子的合成和分配， 了解它们的一般情况，并揭示它们作为抗病毒，抗肿瘤， 治疗在此资助期间，我们开发了一种方法来可视化个别EB病毒的质粒 在整个细胞周期中活细胞中的复制子。这种方法揭示了一种非随机、有效的 EB病毒的分配机制和其DNA合成的内在效率低下。我们将把这个 研究EBV、KSHV和HPV16完整基因组在其细胞中的合成和分配的方法， 天然宿主细胞特别是，我们将描述EBV在其早期阶段的复制特征。 感染原代B细胞，以确定其建立机制，检查合成效率 和分配KSHV作为其末端重复序列的数量的函数，并确定效率 以及HPV 16在两种上皮细胞中的合成和分配方式。这些实验 将有助于确定这些人类肿瘤病毒的合成和分配特征， 可能与人类基因组不同，因此是治疗干预的潜在靶点。

We want to understand functions that are distinctive to human tumor viruses in pre-neoplastic lesions and in tumor cells. These functions are potential targets for specific anti-viral therapies to treat viral associated cancers. We (Drs. Lambert and Sugden) have studied the replication of EBV and HPV for as long as their plasmid replicons have been known and recently extended our work to include KSHV. The goals of our research have been to characterize in detail the synthesis and partitioning of these viral replicons both to understand them in general and to uncover their unique features as targets for anti-viral, anti-tumor therapies. During this funding period we have developed a method to visualize individually EBV's plasmid replicons in live cells throughout a cell cycle. This approach has revealed a non-random, efficient mechanism for EBV's partitioning and an intrinsic inefficiency in its DNA synthesis. We shall extend this approach to study the synthesis and partitioning of intact genomes of EBV, KSHV, and HPV16 in their natural host cells. In particular, we shall characterize the replication of EBV during the early steps of its infection of primary B-cells to identify its mechanism of establishment, examine the efficiencies of synthesis and partitioning of KSHV as a function of the number of its terminal repeats, and determine the efficiencies and modes of the synthesis and of partitioning of HPV16 in two kinds of epithelial cells. These experiments will help to define characteristics of the synthesis and partitioning of these human tumor viruses which are likely distinct from the human genome and thus potential targets for therapeutic intervention.