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

Abstract

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. IMPORTANCE 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.

Genome-Wide Transcriptome Analysis of Human Papillomavirus 16-Infected Primary Keratinocytes Reveals Subtle Perturbations Mostly due to E7 Protein Expression

“…Genes regulated by the JAK/STAT1 signaling axis were downregulated in WT HPV16 rafts ( Fig. 8), which is in line with the literature (33,34). We also observed subtle reductions in STAT1 and STAT2 but not JAK transcript levels, which did not reach significance (data are available in the GEO database under accession number GSE137965).…”

Dysbiosis of Oral Microbiota and Its Effect on Epithelial-Mesenchymal Transition: a Review

“…The F. nucleatum and P. gingivalis attach and invade epithelial cells through specified mechanisms. As per recent findings, Fusobacterium nucleatum utilizes Fusobacterium adhesin A (FadA), Fusobacterium autotransporter protein 2 (Fap2), and fusobacterial outer membrane protein A to bind and invade epithelial cells [96,97]. FadA binds to Ecadherin and promotes attachment and invasion to the host cells.…”

Generation of tonsil organoids as anex vivomodel for SARS-CoV-2 infection

“…Among the four subparts of the oropharynx, the tonsils (named the palatine tonsils or the oropharyngeal tonsils) are composed of a large population of immune cells and thus responsible for the front line of defense against bacteria and viruses that invade the nasal or the oral cavity. Paradoxically, they also serve as a reservoir for infection of diverse viruses including Epstein-Barr virus, adenoviruses, influenza A and B viruses, herpes simplex virus, rhinovirus, enterovirus and even human papillomavirus (Brook, 1987;Chatterjee et al, 2019;Mellin et al, 2002). Recently, it has been reported that the two major genes required for SARS-CoV-2 entry, ACE2 and TMPRSS2, are expressed in the squamous epithelium lining oropharyngeal tonsillar tissue (Hou et al, 2020).…”