HPV E6 and NFX1-123 in differentiation, cell regulation, and cancer

项目概要/摘要： 人乳头瘤病毒（HPV）影响高达75%的成年人，并分为高风险（HR）或低风险（HR）。 风险（LR）基于其与癌症的关联。虽然有两种预防性疫苗针对两种（HR HPV 16，18）或四种（HR HPV 16，18和LR HPV 6，11）HPV基因型，在美国的免疫接种已经 不良的吸收和完成。这使得许多女性和男性继续面临HPV相关癌症的风险， 其中一些发病率正在上升。 HR HPV癌基因E6和E7驱动细胞永生化; HR E6特异性地与 几种内源性蛋白质来失调上皮细胞。在我们自己的研究中，我们发现16 E6与 NFX 1 -123这些蛋白质一起在转录后起作用以增加hTERT的表达， 端粒酶的催化亚单位。端粒酶的激活是细胞永生化的关键，是细胞永生化的关键步骤。 癌症发展，并普遍检测到HPV相关的癌症。我们现在准备研究 由16 E6和NFX 1 -123的协同作用调节的其他基因和细胞途径。 在全基因组表达微阵列和验证研究中，我们确定了几个分化 基因和主分化调节因子Notch 1，如通过NFX 1 -123和16 E6上调。有趣的是， 这种分化途径基因的增加不会导致细胞生长停滞。这些数据使我们处于 研究16 E6和NFX 1 -123在病毒生命周期、细胞、癌症中的联合作用的有利地位 发展和进步。我们的具体目标是：（1）确定基因调控的机制， 16 E6和NFX 1 -123。我们已经确定hTERT的转录后基因调控是一个新的和关键的 16 E6和NFX 1 -123的作用。我们假设NFX 1 -123和16 E6共同发挥作用，使其他细胞的功能失调。 基因，hTERT调控作为我们的工作模型。(2)确定HPV和NFX 1 -123如何影响 平衡上皮细胞的分化和持续的细胞增殖。我们发现NFX 1 -123与16 E6 分化基因和Notch 1的表达增加，这些相同的细胞继续增殖， 文化我们将定义NFX 1 -123、16 E6和Notch 1如何调节上皮结构，并假设 分化和生长停滞途径是解偶联的，以允许细胞生长并支持生产性的生长， 和长期HPV感染，随着时间的推移导致恶性变化。(3)确定NFX 1 -123 表达改变并驱动HPV相关癌症的发展和进展。我们发现， NFX 1 -123在宫颈癌细胞系和30%的患者肿瘤样本中的表达。因此，我们假设， 增加的NFX 1 -123及其下游基因靶点有利于致癌进展。使用颈椎 我们将量化NFX 1 -123的变化，并确定关键点， HPV相关癌症需要增加NFX 1 -123。这些研究将扩大我们对 HPV驱动的肿瘤发生，并帮助确定HPV相关癌症的生物标志物和治疗靶点

Project Summary/Abstract: Human papillomavirus (HPV) affects up to 75% of adults and is categorized as high risk (HR) or low risk (LR) based on its association with cancer. Although there are two preventive vaccines against two (HR HPV 16, 18) or four (HR HPV 16, 18 and LR HPV 6, 11) HPV genotypes, immunizations in the USA have had poor uptake and completion. This leaves many women and men at continued risk of HPV-associated cancers, some of which are increasing in incidence. The HR HPV oncogenes E6 and E7 drive cellular immortalization; HR E6 specifically partners with several endogenous proteins to dysregulate epithelial cells. In our own studies, we found that 16E6 interacts with the NFX1-123. Together, these proteins act post-transcriptionally to increase the expression of hTERT, the catalytic subunit of telomerase. Telomerase activation is critical for cellular immortalization, is a key step in cancer development, and is universally detected in HPV-associated cancers. We are now prepared to study other genes and cellular pathways regulated by the concerted actions of 16E6 and NFX1-123. In whole-genome expression microarray and validation studies, we identified several differentiation genes and a master differentiation regulator, Notch1, as upregulated by NFX1-123 and 16E6. Interestingly, this increase in differentiation pathway genes did not lead to cellular growth arrest. These data put us in a strong position to study the combined roles of 16E6 and NFX1-123 in the viral life cycle, the cell, cancer development and progression. Our specific aims are to: (1) Determine the mechanism of gene regulation by 16E6 and NFX1-123. We have identified post-transcriptional gene regulation of hTERT as a new and critical role for 16E6 and NFX1-123. We hypothesize that NFX1-123 and 16E6 function together to dysregulate other genes, with hTERT regulation as our working model. (2) Determine how HPV and NFX1-123 affect the balance of differentiation and continued cellular proliferation in epithelium. We found NFX1-123 with 16E6 increased expression of differentiation genes and Notch1, and these same cells continued to proliferate in culture. We will define how NFX1-123, 16E6, and Notch1 modulate epithelial architecture, and hypothesize the differentiation and growth arrest pathways are uncoupled to allow cellular growth and support a productive and long-lived HPV infection that leads to malignant changes over time. (3) Determine how NFX1-123 expression changes and drives HPV-associated cancer development and progression. We found increased NFX1-123 in cervical cancer cell lines and in 30% of patient tumor samples. Therefore, we hypothesize that increased NFX1-123, and its downstream gene targets, favors oncogenic progression. Using cervical dysplasia models and patient samples, we will quantify changes in NFX1-123 and identify critical points where increased NFX1-123 is needed in HPV-associated cancers. These studies will expand our understanding of HPV-driven oncogenesis and help to identify biomarkers and therapeutic targets for HPV-associated cancers