Revealing the role of cellular microRNAs in human papillomavirus (HPV) replication in oral keratinocytes

人乳头瘤病毒是双链DNA病毒的一个大家族。高危型人乳头瘤病毒（HPV）亚型HPV 16和HPV 18是通过其病毒癌蛋白E6和E7失调促进上皮细胞癌变的主要病原体。这会导致宫颈癌、肛门生殖道癌和口咽癌。尽管四价HPV疫苗接种计划在预防HPV驱动的宫颈癌方面有效，但HPV现已超过吸烟，成为口咽癌发展的主要致病因素，特别是在男性中。由于宫颈癌发病率高，HPV驱动的头颈癌中的关键病毒靶点尚未得到很好的表征。近年来，对microRNA在一系列癌症发展中的作用的研究加速了; HPV也不例外。microRNA通过其控制基因表达的能力在致癌转化中至关重要。事实上，Macdonald实验室先前发现E6失调的microRNA-18 a是Hippo信号传导中的组成部分，因此促进宫颈癌细胞增殖（Morgan et al.，2020年）。因此，该项目旨在鉴定受HPV操纵的宿主microRNA，以促进口咽部癌症的进展和病毒感染的传播。这将是一个多学科项目，开始通过分析HPV阳性临床头颈部样本与HPV阴性近端组织相比的RNA测序数据。从那里，我们可以努力揭示HPV操纵特定microRNA背后的“方式和原因”，以及它们对促进细胞转化的贡献。该项目还将探索如何利用宿主microRNA来支持使用器官型筏培养的HPV病毒生命周期。这将有可能发展我们对HPV感染背后的细胞机制的理解，这些细胞机制被成功清除，并且变得持久。总之，该项目将确定新的抗病毒靶点，这可能对治疗HPV病毒感染和HPV驱动的致癌作用都很重要。

Human papillomaviruses are a large family of double-stranded DNA viruses. The high-risk human papillomavirus (HPV) subtypes, HPV16 and HPV18, are the main etiological agents in promoting epithelial cell carcinogenesis through dysregulation of its viral oncoproteins E6 and E7. This results in cancers of the cervix, anogenital tract and oropharynx. Despite the effectiveness of the quadrivalent HPV vaccination programme in preventing HPV-driven cervical cancer, HPV has now overtaken smoking as the leading causative agent in the development of oropharyngeal cancers, particularly in men. Due to the high incidence of cervical cancer, key viral targets in HPV-driven head and neck cancers are not well characterised.In recent years, research into the roles of microRNAs in the development of an array of cancers has accelerated; and HPV has been no exception. MicroRNAs are crucial in oncogenic transformation through their ability to control gene expression. Indeed, the Macdonald lab previously uncovered the E6 dysregulated microRNA-18a as an integral component in Hippo signalling and thus for the promotion of cervical cancer cell proliferation (Morgan et al., 2020). Therefore, this project aims to identify host microRNAs that are manipulated by HPV to promote the progression of cancers and propagation of viral infection in the oropharynx. This will be a multidisciplinary project, and begin by analysing RNA-sequencing data of HPV-positive clinical head and neck samples compared to a HPV-negative proximal tissue. From there, we can work to uncover the 'how and why' behind the manipulation of particular microRNAs by HPV, and their contribution to promoting cellular transformation. This project will also explore how host microRNAs are harnessed to support the HPV viral lifecycle using organotypic raft cultures. This will potentially develop our understanding of the cellular mechanisms behind HPV infections that are successfully cleared, and those that become persistent. In summary, this project will identify novel antiviral targets, that may be important in treating both HPV viral infection and HPV-driven carcinogenesis.