Cervical Cancer Development: Implications of HPV16 E6E7-NFX1-123 Regulated Genes.

Kevin M Quist,Nicholas A Wallace,Cen Wu,Isaiah Solorzano,Sreenivasulu Chintala,Rachel A Katzenellenbogen,Sebastian O Wendel

doi:10.3390/cancers13246182

Abstract

Simple SummaryHigh-risk human papillomavirus (HPV) causes 4.5% of cancers and nearly all cervical cancers. HPV’s carcinogenic potential depends on its misappropriation of cellular proteins by HPV’s oncoproteins E6 and E7. High-risk HPV type 16 (HPV16) E6 binds directly to the cellular protein NFX1-123 and dysregulates proliferation, differentiation, and immunity genes. The effect of HPV16 E7 has not been studied in relation to HPV16 E6-NFX1-123-mediated dysregulation. As HPV expresses both oncogenes, and HPV carcinogenesis requires E6 and E7, it is valuable to investigate what dysregulations occur in this context. It is also important to understand their clinical and prognostic ramifications. This study’s goal was to define the gene expression profile regulated by HPV16 E6, E7, and NFX1-123 across cervical precancers and cancers, identify genes correlating with disease progression, assess patient survival, and validate findings in cell models. Finding correlates of survival and disease progression aids in biomarker identification and focuses future studies.High-risk human papillomavirus (HR HPV) causes nearly all cervical cancers, half of which are due to HPV type 16 (HPV16). HPV16 oncoprotein E6 (16E6) binds to NFX1-123, and dysregulates gene expression, but their clinical implications are unknown. Additionally, HPV16 E7’s role has not been studied in concert with NFX1-123 and 16E6. HR HPVs express both oncogenes, and transformation requires their expression, so we sought to investigate the effect of E7 on gene expression. This study’s goal was to define gene expression profiles across cervical precancer and cancer stages, identify genes correlating with disease progression, assess patient survival, and validate findings in cell models. We analyzed NCBI GEO datasets containing transcriptomic data linked with cervical cancer stage and utilized LASSO analysis to identify cancer-driving genes. Keratinocytes expressing 16E6 and 16E7 (16E6E7) and exogenous NFX1-123 were tested for LASSO-identified gene expression. Ten out of nineteen genes correlated with disease progression, including CEBPD, NOTCH1, and KRT16, and affected survival. 16E6E7 in keratinocytes increased CEBPD, KRT16, and SLPI, and decreased NOTCH1. Exogenous NFX1-123 in 16E6E7 keratinocytes resulted in significantly increased CEBPD and NOTCH1, and reduced SLPI. This work demonstrates the clinical relevance of CEBPD, NOTCH1, KRT16, and SLPI, and shows the regulatory effects of 16E6E7 and NFX1-123.

Highlights

Human papillomavirus (HPV) is a small, non-enveloped, double-stranded DNA virus of ~8000 bp that infects mucosal and cutaneous epithelium
NFX1 Expression in Cervical Cancer Correlates with Expression of Genes Involved in Previous publications have demonstrated that elevated expression of NFX1 correPrevious publications have demonstrated that elevated expression of NFX1 correlated lated with changes in the expression of other genes, especially in the context of cells that with changes in the expression of other genes, especially in the context of cells that are are expressing 16 (HPV16). HPV16 oncoprotein E6 (16E6) [23,25]
To identify genes whose expression correlated with NFX1 in cervicervical cancers, we ranked genes based on the extent to which their expression significal cancers, we ranked genes based on the extent to which their expression significantly cantly correlated with NFX1 expression in the Cancer Genome Atlas (TCGA) cervical cancorrelated with NFX1 expression in the Cancer Genome Atlas (TCGA) cervical cancer cer data set (Supplemental Table S2)

Summary

Introduction

Human papillomavirus (HPV) is a small, non-enveloped, double-stranded DNA virus of ~8000 bp that infects mucosal and cutaneous epithelium. The alpha genus is further divided into low-risk types (LR), which can cause benign epithelial lesions such as genital warts, and high-risk types (HR), which can precipitate cervical, anogenital, and head and neck cancers [2]. HR HPV causes 4.5% of all cancers worldwide and virtually every instance of cervical cancer, which is the fourth most common cancer in women globally [7]. HPV+ head and neck cancers are on the rise, especially in North America and northern Europe [9,10]. Of the many HR HPV types, HPV type 16 (HPV16) is responsible for the majority of all HPV+ cancers, with over 50% of cervical cancers and 70% of HPV+ head and neck squamous cell carcinomas attributable to persistent infection with this type [11,12]

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