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. 2018 Sep 26;92(20). doi: 10.1128/JVI.01261-18. Print 2018 Oct 15.
Human papillomavirus (HPV) infection is the world's most common sexually transmitted infection and is responsible for most cases of cervical cancer. Previous studies of global gene expression changes induced by HPV infection have focused on the cancerous stages of infection, and therefore, not much is known about global gene expression changes at early preneoplastic stages of infection. We show for the first time the global gene expression changes during early-stage HPV16 infection in cervical tissue using 3-dimensional organotypic raft cultures, which produce high levels of progeny virions. cDNA microarray analysis showed that a total of 594 genes were upregulated and 651 genes were downregulated at least 1.5-fold with HPV16 infection. Gene ontology analysis showed that biological processes including cell cycle progression and DNA metabolism were upregulated, while skin development, immune response, and cell death were downregulated with HPV16 infection in cervical keratinocytes. Individual genes were selected for validation at the transcriptional and translational levels, including , which was central to the protein association network of immune response genes, and top downregulated genes , , , and In particular, and were shown to be upregulated in cancer, which contrasts with the gene regulation during the productive replication stage. Organotypic raft cultures, which allow full progression of the HPV life cycle, allowed us to identify novel gene modulations and potential therapeutic targets of early-stage HPV infection in cervical tissue. Additionally, our results suggest that early-stage productive infection and cancerous stages of infection are distinct disease states expressing different host transcriptomes. Persistent HPV infection is responsible for most cases of cervical cancer. The transition from precancerous to cancerous stages of HPV infection is marked by a significant reduction in virus production. Most global gene expression studies of HPV infection have focused on the cancerous stages. Therefore, little is known about global gene expression changes at precancerous stages. For the first time, we measured global gene expression changes at the precancerous stages of HPV16 infection in human cervical tissue producing high levels of virus. We identified a group of genes that are typically overexpressed in cancerous stages to be significantly downregulated at the precancerous stage. Moreover, we identified significantly modulated genes that have not yet been studied in the context of HPV infection. Studying the role of these genes in HPV infection will help us understand what drives the transition from precancerous to cancerous stages and may lead to the development of new therapeutic targets.
人乳头瘤病毒（HPV）感染是世界上最常见的性传播感染，也是大多数宫颈癌的病因。之前关于 HPV 感染引起的全球基因表达变化的研究主要集中在感染的癌变阶段，因此，对于感染早期癌前阶段的全球基因表达变化知之甚少。我们首次使用产生高水平病毒粒子的 3 维器官型筏培养物显示了 HPV16 感染早期阶段宫颈组织中的全球基因表达变化。cDNA 微阵列分析显示，HPV16 感染使总共 594 个基因上调和 651 个基因下调至少 1.5 倍。基因本体论分析显示，包括细胞周期进程和 DNA 代谢在内的生物学过程上调，而皮肤发育、免疫反应和细胞死亡下调。选择个别基因在转录和翻译水平进行验证，包括在免疫反应基因的蛋白质关联网络中起核心作用的 ，以及下调最明显的基因 、 、 和 。特别是 和 在癌症中上调，这与生产性复制阶段的基因调控形成对比。允许 HPV 生命周期完全进展的器官型筏培养物使我们能够识别宫颈组织中早期 HPV 感染的新基因调节和潜在治疗靶点。此外，我们的结果表明，早期生产性感染和癌变阶段是表达不同宿主转录组的不同疾病状态。持续性 HPV 感染是大多数宫颈癌的病因。从癌前阶段到 HPV 感染的癌变阶段的转变以病毒产量显著减少为标志。大多数 HPV 感染的全球基因表达研究都集中在癌变阶段。因此，对于癌前阶段的全球基因表达变化知之甚少。我们首次测量了 HPV16 感染在产生高水平病毒的人类宫颈组织中的癌前阶段的全球基因表达变化。我们确定了一组在癌变阶段通常过表达的基因在癌前阶段显著下调。此外，我们还确定了在 HPV 感染背景下尚未研究过的显著调节基因。研究这些基因在 HPV 感染中的作用将帮助我们了解是什么驱动了从癌前到癌变阶段的转变，并可能导致新的治疗靶点的发展。
