Department of Microbiology and Immunology, Penn State College of Medicine, Hershey, Pennsylvania, USA.
Bioinformatics Core, Penn State College of Medicine, Hershey, Pennsylvania, USA.
J Virol. 2019 Aug 13;93(17). doi: 10.1128/JVI.00915-19. Print 2019 Sep 1.
Epidemiological data confirm a much higher incidence of high-risk human papillomavirus 16 (HPV16)-mediated carcinogenesis of the cervical epithelium than for other target sites. In order to elucidate tissue-specific responses to virus infection, we compared gene expression changes induced by productive HPV16 infection of cervical, foreskin, and tonsil organotypic rafts. These rafts closely mimic persistent HPV16 infection, long before carcinogenesis sets in. The total number of gene expression changes varied considerably across the tissue types, with only 32 genes being regulated in common. Among them, we confirmed the Kelch-like family protein KLHL35 and the laminin-5 complex to be upregulated and downregulated, respectively, in all the three tissues. HPV16 infection induces upregulation of genes involved in cell cycle control, cell division, mitosis, DNA replication, and DNA damage repair in all the three tissues, indicative of a hyperproliferative environment. In the cervical and tonsil epithelium, we observe significant downregulation of genes involved in epidermis development, keratinocyte differentiation, and extracellular matrix organization. On the other hand, in HPV16-positive foreskin (HPV16 foreskin) tissue, several genes involved in interferon-mediated innate immunity, cytokine signaling, and cellular defenses were downregulated. Furthermore, pathway analysis and experimental validations identified important cellular pathways like STAT1 and transforming growth factor β (TGF-β) to be differentially regulated among the three tissue types. The differential modulation of important cellular pathways like TGF-β1 and STAT1 can explain the sensitivity of tissues to HPV cancer progression. Although the high-risk human papillomavirus 16 infects anogenital and oropharyngeal sites, the cervical epithelium has a unique vulnerability to progression of cancer. Host responses during persistent infection and preneoplastic stages can shape the outcome of cancer progression in a tissue-dependent manner. Our study for the first time reports differential regulation of critical cellular functions and signaling pathways during productive HPV16 infection of cervical, foreskin, and tonsil tissues. While the virus induces hyperproliferation in infected cells, it downregulates epithelial differentiation, epidermal development, and innate immune responses, according to the tissue type. Modulation of these biological functions can determine virus fitness and pathogenesis and illuminate key cellular mechanisms that the virus employs to establish persistence and finally initiate disease progression.
流行病学数据证实，高危型人乳头瘤病毒 16（HPV16）介导的宫颈上皮癌变的发生率远高于其他靶位。为了阐明组织对病毒感染的特异性反应，我们比较了 HPV16 感染宫颈、包皮和扁桃体器官样筏时诱导的基因表达变化。这些筏在致癌作用开始之前，非常接近持续 HPV16 感染。组织类型之间的基因表达变化数量差异很大，只有 32 个基因共同调节。其中，我们证实 Kelch 样家族蛋白 KLHL35 和层粘连蛋白 5 复合物在所有三种组织中分别上调和下调。HPV16 感染诱导所有三种组织中参与细胞周期控制、细胞分裂、有丝分裂、DNA 复制和 DNA 损伤修复的基因上调，表明存在过度增殖的环境。在宫颈和扁桃体上皮中，我们观察到参与表皮发育、角质形成细胞分化和细胞外基质组织的基因显著下调。另一方面，在 HPV16 阳性包皮（HPV16 包皮）组织中，参与干扰素介导的先天免疫、细胞因子信号和细胞防御的几个基因下调。此外，途径分析和实验验证确定了重要的细胞途径，如 STAT1 和转化生长因子β（TGF-β）在三种组织类型中存在差异调节。重要细胞途径如 TGF-β1 和 STAT1 的差异调节可以解释组织对 HPV 癌症进展的敏感性。虽然高危型人乳头瘤病毒 16 感染肛门生殖器和口咽部位，但宫颈上皮对癌症进展具有独特的易感性。持续性感染和癌前阶段的宿主反应可以以组织依赖的方式影响癌症进展的结果。我们的研究首次报告了 HPV16 在宫颈、包皮和扁桃体组织中感染时关键细胞功能和信号通路的差异调节。虽然病毒在感染细胞中诱导过度增殖，但根据组织类型，它下调上皮分化、表皮发育和先天免疫反应。这些生物学功能的调节可以决定病毒的适应性和发病机制，并阐明病毒用来建立持久性并最终引发疾病进展的关键细胞机制。
