Single cell transcriptomic analysis of HPV16-infected epithelium identifies a keratinocyte subpopulation implicated in cancer

Abstract

Persistent HPV16 infection is a major cause of the global cancer burden. The viral life cycle is dependent on the differentiation program of stratified squamous epithelium, but the landscape of keratinocyte subpopulations which support distinct phases of the viral life cycle has yet to be elucidated. Here, single cell RNA sequencing of HPV16 infected compared to uninfected organoids identifies twelve distinct keratinocyte populations, with a subset mapped to reconstruct their respective 3D geography in stratified squamous epithelium. Instead of conventional terminally differentiated cells, an HPV-reprogrammed keratinocyte subpopulation (HIDDEN cells) forms the surface compartment and requires overexpression of the ELF3/ESE-1 transcription factor. HIDDEN cells are detected throughout stages of human carcinogenesis including primary human cervical intraepithelial neoplasias and HPV positive head and neck cancers, and a possible role in promoting viral carcinogenesis is supported by TCGA analyses. Single cell transcriptome information on HPV-infected versus uninfected epithelium will enable broader studies of the role of individual keratinocyte subpopulations in tumor virus infection and cancer evolution.

The macrophage polarization in Entamoeba histolytica infection modulation by the C fragment of the intermediate subunit of Gal/GalNAc-inhibitable lectin

“…Consequently, we hypothesized that Igl-C is rich in CXXC motifs, which can alter the disulfide bonds between the surface proteins of pathogens and promote the formation, reduction, and rearrangement of disulfide bonds ( 15 ), resulting in antigenic variation, inhibition of the inflammatory response, and ultimately persistent infection of pathogens ( 10 , 31 , 32 ). Single-cell transcriptomics has been used to study the functional diversity changes in macrophages in persistent infection status ( 33 ). Factors that influence macrophage heterogeneity in the context of persistent infection have been identified, and the various phenotypes, functional programming, and spatial distributions of these cells have been described.…”

Targeting CD74-positive macrophages improves neoadjuvant therapy in cervical cancer as revealed by single-cell transcriptomics analysis

“…Previous scRNA-seq studies focus on the heterogeneity of human cervical squamous cell carcinoma initiation and progression and capture the overall picture 27,28 . Specific subpopulations of cervical cancer, such as the HPV-reprogrammed keratinocyte subpopulation, were identified to provide evidence for tumor virus infection and cancer evolution 29 . The radiochemotherapy-induced innate immune activation and MHC-II upregulation in cervical cancer was also revealed by scRNA-seq 30 .…”

Structure and transcription of integrated HPV DNA in vulvar carcinomas

“…To visualize genomic regions of interest we used Bacterial Artificial Chromosome (BAC) clones RP11-990E14 (hg38 -chr3:169,692,858-169,870,368 mapping to Telomerase RNA Component TERC ), and RP11-662P23 (hg38 - chr5:56,420,151-56,608,998 mapping to the specific region of HPV16 integration on chromosome 5 in Tumor 4). In addition to human BAC clones we also used a plasmid containing the entire genomic sequence of HPV16 as reported in 100 , 101 . BAC DNA and HPV16 plasmid DNA was isolated and labeled by nick translation as we previously described 102 , 103 using the following dUTPs from Dyomics (Jena, GE, USA): DY-495-dUTP (within the green spectrum to visualize RP11-662P23); DY-530-aadUTP (within the gold spectrum to visualize RP11-990E14); and DY-590-dUTP (within the red spectrum to visualize HPV16).…”