High-risk HPV E6: dysregulation of immortalization, growth, and differentiation through protein partnerships in HPV-associated cancers

项目摘要/摘要： 高危人类乳头瘤病毒(HR HPV)导致了5%的癌症。有预防性疫苗 针对HPV，但只有不到2%的世界目标人口接种了这种疫苗。这就留下了数百万 女性和男性面临HPV相关的宫颈癌、肛门癌和头颈癌的风险。我们需要 了解HR HPV如何建立和维持活跃的感染，同时也推动癌症的发展， 以指导有针对性的治疗干预措施，以预防、遏制和逆转疾病。 宫颈癌的最大临床风险因素是持续的HR HPV感染。这种坚持不懈 通过细胞通路的协调失调而发生，细胞通路既支持感染又促进 癌症的发展。这种失调的背后是要求HPV癌基因E6和E7 与细胞蛋白质配对。我们的工作主要集中在HPV16型(16E6)的E6癌基因上， 癌症中常见的HR-HPV型。此前，我们发现16E6需要细胞蛋白NFX1-123，并且 它的蛋白质伙伴胞浆聚(A)结合蛋白(PABPC)，以充分激活端粒酶和 永生之路。我们目前的研究表明，NFX1-123在宫颈癌中高度表达， 随着时间的推移，16E6、NFX1-123和PABPC共同放大端粒酶、细胞生长和寿命。我们 还发现NFX1-123在分化过程中增加，并与16E6一起增强细胞 分化级联及其宿主和病毒基因靶点同时保护 伴随而来的是细胞停滞和衰老。这些发现为新的调查水平创造了一个视线 这将揭示增长和分化的连通性和控制性，以及推动时间变化的因素 以及PABPC、NFX1-123和16E6加速永生。这些结果还将勾勒出目标 用于未来的治疗，特别是扰乱HPV及其癌症所需的通用途径。 我们的具体目标是：(1)确定16E6、NFX1-123和PABPC如何在共同调节中发挥作用 生长和分化，以更好地建立持续感染。我们将模仿建立 HR HPV感染以确定这些蛋白质允许分化和生长的功能方式 在HR HPV感染的最初、基础步骤中协调一致。(2)阐明心绞痛的发病机制 16E6、NFX1-123和PABPC纵向、顺序增加端粒酶和端粒酶。端粒酶 激活会导致细胞永生。我们将利用长期的细胞研究来确定 16E6与NFX1-123和PABPC对端粒酶催化亚基hTERT序列的影响 并创建反映临床年表的分子致癌进展路线图。我们的建议 研究将阐明16E6及其宿主对致癌途径的时间和相互交织的失调 蛋白质合作伙伴；他们还将提供关于肿瘤病因和进展的基础数据 越来越常见的HPV相关癌症。

Project Summary/Abstract: High-risk human papillomaviruses (HR HPVs) cause 5% of all cancers. There are preventive vaccines against HPV, but less than 2% of the world’s target population has received them. This leaves millions of women and men at risk for HPV-associated cervical, anogenital, and head and neck cancers. We need to understand how HR HPV establishes and maintains an active infection, while also driving cancer development, in order to guide focused therapeutic interventions to prevent, arrest, and reverse disease. The greatest clinical risk factor for cervical cancer is a persistent HR HPV infection. This persistence occurs through coordinated dysregulation of cellular pathways, which both support the infection and foster cancer development. Underlying this dysregulation is the requirement that the HPV oncogenes E6 and E7 partner with cellular proteins. Our work has focused on the E6 oncogene from HPV type 16 (16E6), the most common HR HPV type in cancers. Previously, we found that 16E6 required the cellular protein NFX1-123, and its protein partners cytoplasmic poly(A) binding proteins (PABPCs), to fully activate telomerase and the immortalization pathway. Our current studies revealed that NFX1-123 is highly expressed in cervical cancers, and together 16E6, NFX1-123, and PABPCs amplify telomerase, cellular growth, and longevity over time. We also discovered that NFX1-123 is increased during differentiation and, together with 16E6, augments cellular differentiation cascades and their host and viral gene targets while simultaneously protecting against concomitant cellular arrest and senescence. These findings create a sightline for a new level of investigation that will uncover the connectivity and control of growth and differentiation and the temporal changes driving and accelerating immortalization by PABPCs, NFX1-123, and 16E6. These results will also delineate targets for future treatments that specifically disrupt universal pathways required for HPV and its cancers. Our specific aims are: (1) Determine how 16E6, NFX1-123, and PABPCs work together in co-regulating growth and differentiation to better establish a persistent infection. We will mimic the initial steps of establishing a HR HPV infection to identify the way in which these proteins function to permit both differentiation and growth in concert during the initial, foundational steps of a HR HPV infection. (2) Elucidate the mechanism of longitudinal, sequential increases of hTERT and telomerase by 16E6, NFX1-123, and PABPCs. Telomerase activation leads to cellular immortalization. We will leverage long-term cellular studies to determine the sequential changes to hTERT, the catalytic subunit of telomerase, due to 16E6 with NFX1-123 and PABPCs and to create a roadmap of molecular oncogenic progression that mirrors clinical chronology. Our proposed studies will elucidate the temporal and interwoven dysregulation of oncogenic pathways by 16E6 and its host protein partners; they will also provide foundational data on the oncogenic etiology and progression of increasingly common HPV-associated cancers.