Effect of Productive Human Papillomavirus 16 Infection on Global Gene Expression in Cervical Epithelium

Abstract

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.

Tissue-Specific Gene Expression during Productive Human Papillomavirus 16 Infection of Cervical, Foreskin, and Tonsil Epithelium

“…We have previously shown that KLK8 and RPTN are downregulated during productive HPV16 infection of cervix (25). We confirmed the exclusive downregulation of KLK8 and RPTN mRNAs in HPV16 cervical tissue (Fig.…”

“…Our previous publication discusses the gene expression changes during productive HPV16 infection of the cervical tissue (25). In this paper, we perform a comparative analysis of gene expression changes induced by productive HPV16 infection of cervical, foreskin, and tonsil tissues.…”

Expression of Viral microRNAs in Serum and White Blood Cells of Cows Exposed to Bovine Leukemia Virus

“…Blv-miR-B2-5p was found to be associated with the expression of RPTN. This gene is involved in cell differentiation and is down-regulated in patients with human papillomavirus (29). It is probable that blv-miR-B2-5p may down-regulate RPTN during a BLV infection.…”

Suppression of Stromal Interferon Signaling by Human Papillomavirus 16

“…Our results indicate that many genes and pathways are regulated by HPV in the stroma, consistent with the wide array of soluble factors dysregulated in infected keratinocytes by the viral oncogenes (48-50, 54, 80, 81). Most previous studies related to global gene expression patterns during HPV infection have focused on the effect of HPV infection on the epithelium, and mostly as relating to carcinogenesis (49,54,(82)(83)(84)(85)(86)(87)(88). Some gene expression profiling studies have considered stromal changes in the context of HPV infection and cancer development (53,54,88,89), but a clear picture of how the stroma impacts HPV (and vice versa) during benign productive infection is lacking.…”