ABSTRACT
Objective: The objective of the study is to validate a new human papillomavirus (HPV) L1 high-risk specific serological assay in a case-control study. Methods: Serum samples of 138 patients (cervical intraepithelial neoplasia (CIN) 1, 2, and 3 and cervical cancer), 21 vaccinees, and 246 female controls were tested for the presence of HPV L1 high-risk specific antibodies. Results: HPV L1 high-risk antibodies were detected in 100% of the CIN1 and 2, 86.6% of the CIN3 and 82.4% of the cervical cancer cases, 100% of the vaccinees, and 3.9% of the female controls. Area under the curve (AUC) was calculated with 0.91 for controls versus CIN2+, 0.923 for controls versus CIN1+, and 0.968 for controls versus CIN1/2. Conclusion: The HPV L1 high-risk specific serological lateral flow rapid test shows promising data in the field of early detection of HPV high-risk induced cervical cancer and its precursor lesions. This easy-to-use, robust, and affordable approach could offer a chance to reach women in low- or middle-income countries (LMICs) that could not be reached by HPV molecular testing-based cervical cancer screening programs.
Subject(s)
Antibodies, Viral , Early Detection of Cancer , Papillomavirus Infections , Sensitivity and Specificity , Uterine Cervical Dysplasia , Uterine Cervical Neoplasms , Humans , Female , Uterine Cervical Neoplasms/virology , Uterine Cervical Neoplasms/diagnosis , Case-Control Studies , Adult , Papillomavirus Infections/diagnosis , Papillomavirus Infections/virology , Antibodies, Viral/blood , Uterine Cervical Dysplasia/virology , Uterine Cervical Dysplasia/diagnosis , Middle Aged , Early Detection of Cancer/methods , Oncogene Proteins, Viral/immunology , Capsid Proteins/immunology , Young Adult , Papillomaviridae/immunology , Papillomaviridae/isolation & purification , AgedABSTRACT
Human papillomaviruses (HPVs) account for more than 30% of cancer cases, with definite identification of the oncogenic role of viral E6 and E7 genes. However, the identification of high-risk HPV genotypes has largely relied on lagged biological exploration and clinical observation, with types unclassified and oncogenicity unknown for many HPVs. In the present study, we retrieved and cleaned HPV sequence records with high quality and analyzed their genomic compositional traits of dinucleotide (DNT) and DNT representation (DCR) to overview the distribution difference among various types of HPVs. Then, a deep learning model was built to predict the oncogenic potential of all HPVs based on E6 and E7 genes. Our results showed that the main three groups of Alpha, Beta, and Gamma HPVs were clearly separated between/among types in the DCR trait for either E6 or E7 coding sequence (CDS) and were clustered within the same group. Moreover, the DCR data of either E6 or E7 were learnable with a convolutional neural network (CNN) model. Either CNN classifier predicted accurately the oncogenicity label of high and low oncogenic HPVs. In summary, the compositional traits of HPV oncogenicity-related genes E6 and E7 were much different between the high and low oncogenic HPVs, and the compositional trait of the DCR-based deep learning classifier predicted the oncogenic phenotype accurately of HPVs. The trained predictor in this study will facilitate the identification of HPV oncogenicity, particularly for those HPVs without clear genotype or phenotype.
Subject(s)
Deep Learning , Genome, Viral , Papillomaviridae , Papillomavirus Infections , Humans , Papillomavirus Infections/virology , Papillomaviridae/genetics , Genome, Viral/genetics , Genotype , Oncogene Proteins, Viral/genetics , Papillomavirus E7 Proteins/genetics , Carcinogenesis/geneticsABSTRACT
Human papillomavirus (HPV) 11/16 E6/E7 proteins have been recognized to be pivotal in viral pathogenesis. This study sought to uncover the potential mechanisms of how HPV11/16 E6/E7-transfected keratinocytes inhibit cytokine secretion in peripheral blood mononuclear cells (PBMC). Upon co-culturing HPV11/16 E6/E7-transfected keratinocytes with PBMC in a non-contact manner, we observed a marked decrease in various cytokines secreted by PBMC. To determine if this suppression was mediated by specific common secreted factors, we conducted transcriptomic sequencing on these transfected cells. This analysis identified 53 common differentially secreted genes in all four HPV-transfected cells. Bioinformatics analysis demonstrated these genes were predominantly involved in immune regulation. Results from quantitative PCR (qPCR) and an extensive literature review suggested the downregulation of 12 genes (ACE2, BMP3, BPIFB1, CLU, CST6, CTF1, HMGB2, MMP12, PDGFA, RNASE7, SULF2, TGM2), and upregulation of 7 genes (CCL17, CCL22, FBLN1, PLAU, S100A7, S100A8, S100A9), may be crucial in modulating tumor immunity and combating pathogenic infections, with genes S100A8 and S100A9, and IL-17 signaling pathway being particularly noteworthy. Thus, HPV11/16 E6/E7 proteins may inhibit cytokine secretion of immune cells by altering the expression of host-secreted genes. Further exploration of these genes may yield new insights into the complex dynamics of HPV infection.
Subject(s)
Cytokines , Leukocytes, Mononuclear , Oncogene Proteins, Viral , Humans , Cytokines/metabolism , Cytokines/genetics , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/virology , Leukocytes, Mononuclear/metabolism , Oncogene Proteins, Viral/genetics , Oncogene Proteins, Viral/metabolism , Oncogene Proteins, Viral/immunology , Keratinocytes/virology , Keratinocytes/immunology , Keratinocytes/metabolism , Human papillomavirus 16/genetics , Human papillomavirus 16/immunology , Human papillomavirus 11/genetics , Human papillomavirus 11/immunology , Gene Expression Profiling , Papillomavirus Infections/virology , Papillomavirus Infections/immunology , Papillomavirus Infections/genetics , Papillomavirus E7 Proteins/genetics , Papillomavirus E7 Proteins/metabolism , Papillomavirus E7 Proteins/immunology , Coculture Techniques , Host-Pathogen Interactions/immunology , Host-Pathogen Interactions/geneticsABSTRACT
Human papilloma virus (HPV) is an etiological factor of head and neck squamous cell carcinoma (HNSCC). To investigate the role of HPV antigen in anti-tumor immunity, we established mouse models by expressing HPV16 E6 and E7 in a SCC tumor cell line. We obtained two HPV antigen-expressing clones (C-225 and C-100) transplantable into C57BL/6 recipients. We found that C-225 elicited complete eradication in C57BL/6 mice (eradicated), whereas C-100 grew progressively (growing). We examined immune tumor microenvironment (TME) using flow cytometry and found that eradicated or growing tumors exhibited differential immune profiles that may influence the outcome of anti-tumor immunity. Surprisingly, the percentage of CD8 and CD4 tumor-infiltrating lymphocytes (TILs) was much higher in growing (C-100) than eradicated (C-225) tumor. However, the TILs upregulated PD-1 and LAG-3 more potently and exhibited impaired effector functions in growing tumor compared to their counterparts in eradicated tumor. C-225 TME is highly enriched with myeloid cells, especially polymorphonuclear (PMN) myeloid-derived suppressor cells (MDSC), whereas the percentage of M-MDSC and tumor-associated macrophages (TAMs) was much higher in C-100 TME, especially M2-TAMs (CD206+). The complete eradication of C-225 depended on CD8 T cells and elicited anti-tumor memory responses upon secondary tumor challenge. We employed DNA sequencing to identify differences in the T cell receptor of peripheral blood lymphocytes pre- and post-secondary tumor challenge. Lastly, C-225 and C-100 tumor lines harbored different somatic mutations. Overall, we uncovered differential immune TME that may underlie the divergent outcomes of anti-tumor immunity by establishing two SCC tumor lines, both of which express HPV16 E6 and E7 antigens. Our experimental models may provide a platform for pinpointing tumor-intrinsic versus host-intrinsic differences in orchestrating an immunosuppressive TME in HNSCCs and for identifying new targets that render tumor cells vulnerable to immune attack.
Subject(s)
Disease Models, Animal , Lymphocytes, Tumor-Infiltrating , Mice, Inbred C57BL , Oncogene Proteins, Viral , Papillomavirus E7 Proteins , Papillomavirus Infections , Tumor Microenvironment , Animals , Tumor Microenvironment/immunology , Mice , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Cell Line, Tumor , Oncogene Proteins, Viral/immunology , Oncogene Proteins, Viral/genetics , Papillomavirus E7 Proteins/immunology , Papillomavirus E7 Proteins/genetics , Papillomavirus Infections/immunology , Squamous Cell Carcinoma of Head and Neck/immunology , Squamous Cell Carcinoma of Head and Neck/virology , Repressor Proteins/genetics , Lymphocyte Activation Gene 3 Protein , Humans , Disease Progression , CD8-Positive T-Lymphocytes/immunology , Programmed Cell Death 1 Receptor , Female , Human papillomavirus 16/immunology , Head and Neck Neoplasms/immunology , Head and Neck Neoplasms/virologyABSTRACT
Despite therapeutic advancements, cervical cancer caused by high-risk subtypes of the human papillomavirus (HPV) remains a leading cause of cancer-related deaths among women worldwide. This study aimed to discover potential drug candidates from the Asian medicinal plant Andrographis paniculata, demonstrating efficacy against the E6 protein of high-risk HPV-16 subtype through an in-silico computational approach. The 3D structures of 32 compounds (selected from 42) derived from A. paniculata, exhibiting higher binding affinity, were obtained from the PubChem database. These structures underwent subsequent analysis and screening based on criteria including binding energy, molecular docking, drug likeness and toxicity prediction using computational techniques. Considering the spectrometry, pharmacokinetic properties, docking results, drug likeliness, and toxicological effects, five compounds-stigmasterol, 1H-Indole-3-carboxylic acid, 5-methoxy-, methyl ester (AP7), andrographolide, apigenin and wogonin-were selected as the potential inhibitors against the E6 protein of HPV-16. We also performed 200 ns molecular dynamics simulations of the compounds to analyze their stability and interactions as protein-ligand complexes using imiquimod (CID-57469) as a control. Screened compounds showed favorable characteristics, including stable root mean square deviation values, minimal root mean square fluctuations and consistent radius of gyration values. Intermolecular interactions, such as hydrogen bonds and hydrophobic contacts, were sustained throughout the simulations. The compounds displayed potential affinity, as indicated by negative binding free energy values. Overall, findings of this study suggest that the selected compounds have the potential to act as inhibitors against the E6 protein of HPV-16, offering promising prospects for the treatment and management of CC.
Subject(s)
Andrographis , Human papillomavirus 16 , Molecular Docking Simulation , Oncogene Proteins, Viral , Uterine Cervical Neoplasms , Uterine Cervical Neoplasms/drug therapy , Uterine Cervical Neoplasms/virology , Humans , Female , Oncogene Proteins, Viral/metabolism , Oncogene Proteins, Viral/chemistry , Andrographis/chemistry , Human papillomavirus 16/drug effects , Repressor Proteins/metabolism , Repressor Proteins/antagonists & inhibitors , Repressor Proteins/chemistry , Phytochemicals/pharmacology , Phytochemicals/chemistry , Computer Simulation , Plant Extracts/chemistry , Plant Extracts/pharmacology , Molecular Dynamics Simulation , Papillomavirus Infections/drug therapy , Papillomavirus Infections/virology , Diterpenes/pharmacology , Diterpenes/chemistry , Protein BindingABSTRACT
This study was designed to investigate the expression of HPV16 L1-protein in biopsies of oral mucosa samples. The expression of HPV16 L1 protein was investigated in biopsies taken from oral mucosa from patients who required pathological diagnosis of oral lesions. Seventy-two samples were incubated with anti-L1 protein monoclonal antibodies and protein detection was revealed with diaminobenzidine. Expression of L1 protein was performed by a pathologist blinded for tissue diagnosis under light microscopy. Most of the lesions of oral mucosa were present in lining mucosa (75 %) and the most frequent lesion were mucocele (n = 17, 23.6 %), epithelial hyperplasia (n = 6, 8.33 %), fibroma (n = 5, 6.9 %) and inflammatory hyperplasia (n = 5, 6.9 %). L1 protein expression was observed only in five (6.9 %) samples (two squamous cell carcinomas, two epithelial hyperplasia, and one gingival hyperplasia). We concluded that L1 expression in oral biopsies presented a low frequency in oral mucosal biopsies samples.
Subject(s)
Capsid Proteins , Mouth Mucosa , Oncogene Proteins, Viral , Papillomavirus Infections , Humans , Biopsy , Female , Mouth Mucosa/virology , Mouth Mucosa/pathology , Papillomavirus Infections/diagnosis , Papillomavirus Infections/virology , Adult , Male , Oncogene Proteins, Viral/genetics , Middle Aged , Ecuador/epidemiology , Capsid Proteins/genetics , Capsid Proteins/immunology , Young Adult , Adolescent , Aged , Prevalence , Mouth Diseases/virology , Mouth Diseases/pathology , Mouth Diseases/epidemiology , Human papillomavirus 16/genetics , Immunohistochemistry , Mouth Neoplasms/virology , Mouth Neoplasms/pathology , Mouth Neoplasms/diagnosisABSTRACT
Tirbanibulin 1% ointment is a synthetic antiproliferative agent approved in 2021 by the European Union for treating actinic keratoses (AK). Topical tirbanibulin has clinically resolved HPV-57 ( +) squamous cell carcinoma (SCC), HPV-16 ( +) vulvar high-grade squamous intraepithelial lesion, epidermodysplasia verruciformis, and condyloma. We examined how tirbanibulin might affect HPV oncoprotein expression and affect other cellular pathways involved in cell proliferation and transformation. We treated the HeLa cell line, containing integrated HPV-18, with increasing doses of tirbanibulin to determine the effects on cell proliferation. Immunoblotting was performed with antibodies against the Src canonical pathway, HPV 18 E6 and E7 transcription regulation, apoptosis, and invasion and metastasis pathways. Cell proliferation assays with tirbanibulin determined the half-maximal inhibitory concentration (IC50) of HeLa cells to be 31.49 nmol/L. Increasing concentrations of tirbanibulin downregulates the protein expression of Src (p < 0.001), phospho-Src (p < 0.001), Ras (p < 0.01), c-Raf (p < 0.001), ERK1 (p < 0.001), phospho-ERK1 (p < 0.001), phospho-ERK2 (p < 0.01), phospho-Mnk1 (p < 0.001), eIF4E (p < 0.01), phospho-eIF4E (p < 0.001), E6 (p < 0.01), E7 (p < 0.01), Rb (p < 0.01), phospho-Rb (p < 0.001), MDM2 (p < 0.01), E2F1 (p < 0.001), phospho-FAK (p < 0.001), phospho-p130 Cas (p < 0.001), Mcl-1 (p < 0.01), and Bcl-2 (p < 0.001), but upregulates cPARP (p < 0.001), and cPARP/fPARP (p < 0.001). These results demonstrate that tirbanibulin may impact expression of HPV oncoproteins via the Src- MEK- pathway. Tirbanibulin significantly downregulates oncogenic proteins related to cell cycle regulation and cell proliferation while upregulating apoptosis pathways.
Tirbanibulin is Promising Novel Therapy for Human Papillomavirus (HPV)-associated Diseases.Tirbanibulin 1% ointment is an approved synthetic topical ointment for treating actinic keratoses (AK), a precancer of skin cancer. Topical tirbanibulin has previously been reported to clinically resolve human papillomavirus (HPV)-( +) diseases.In this study, we examine how tirbanibulin may affect the HPV and pathways associated with cancer.We treated the HeLa cell line to determine the effects on HPV cell proliferation. Increasing the concentration of tirbanibulin statistically significantly affected numerous cellular pathways often associated with cancer.These results demonstrate that tirbanibulin may impact expression of HPV oncoproteins and thereby kill cancer cells.
Subject(s)
Cell Proliferation , Down-Regulation , Human papillomavirus 18 , Oncogene Proteins, Viral , Humans , HeLa Cells , Cell Proliferation/drug effects , Oncogene Proteins, Viral/metabolism , Down-Regulation/drug effects , Papillomavirus Infections/virology , Papillomavirus Infections/drug therapy , Papillomavirus E7 Proteins/metabolism , Apoptosis/drug effects , Repressor Proteins/metabolism , Repressor Proteins/genetics , Signal Transduction/drug effects , Uterine Cervical Neoplasms/virology , Uterine Cervical Neoplasms/drug therapy , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/metabolism , src-Family Kinases/metabolism , src-Family Kinases/antagonists & inhibitors , Female , Human Papillomavirus Viruses , DNA-Binding ProteinsABSTRACT
BACKGROUND: There is a high incidence of cervical cancer in Xinjiang. Genetic variation in human papillomavirus may increase its ability to invade, spread, and escape host immune response. METHODS: HPV16 genome was sequenced for 90 positive samples of HPV16 infection. Sequences of the E4, E5 and L2 genes were analysed to reveal sequence variation of HPV16 in Xinjiang and the distribution of variation among the positive samples of HPV16 infection. RESULTS: Eighty-one of the 90 samples of HPV16 infection showed variation in HPV16 E4 gene with 18 nucleotide variation sites, of which 8 sites were synonymous variations and 11 missense variations. 90 samples of HPV16 infection showed variation in HPV16 E5 and L2 genes with 16 nucleotide variation sites (6 synonymous, 11 missense variations) in the E5 gene and 100 nucleotide variation sites in L2 gene (37 synonymous, 67 missense variations). The frequency of HPV16 L2 gene missense variations G3377A, G3599A, G3703A, and G3757A was higher in the case groups than in the control groups. CONCLUSIONS: Phylogenetic tree analysis showed that 87 samples were European strains, 3 cases were Asian strains, there were no other variations, and G4181A was related to Asian strains. HPV16 L2 gene missense variations G3377A, G3599A, G3703A, and G3757A were significantly more frequent in the case groups than in the control groups.
Subject(s)
Genetic Variation , Human papillomavirus 16 , Oncogene Proteins, Viral , Papillomavirus Infections , Phylogeny , Humans , Female , China , Human papillomavirus 16/genetics , Papillomavirus Infections/virology , Papillomavirus Infections/genetics , Oncogene Proteins, Viral/genetics , Uterine Cervical Neoplasms/virology , Uterine Cervical Neoplasms/genetics , Adult , Middle Aged , Mutation, MissenseABSTRACT
BACKGROUND: High-risk human papillomavirus (HR-HPV) infection is an important factor for the development of cervical cancer. HPV18 is the second most common HR-HPV after HPV16. METHODS: In this study, MEGA11 software was used to analyze the variation and phylogenetic tree of HPV18 E6-E7 and L1 genes. The selective pressure to E6, E7 and L1 genes was estimated using pamlX. In addition, the B cell epitopes of L1 amino acid sequences and T cell epitopes of E6-E7 amino acid sequences in HPV18 were predicted by ABCpred server and IEDB website, respectively. RESULTS: A total of 9 single nucleotide variants were found in E6-E7 sequences, of which 2 were nonsynonymous variants and 7 were synonymous variants. Twenty single nucleotide variants were identified in L1 sequence, including 11 nonsynonymous variants and 9 synonymous variants. Phylogenetic analysis showed that E6-E7 and L1 sequences were all distributed in A lineage. In HPV18 E6, E7 and L1 sequences, no positively selected site was found. The nonconservative substitution R545C in L1 affected hypothetical B cell epitope. Two nonconservative substitutions, S82A in E6, and R53Q in E7, impacted multiple hypothetical T cell epitopes. CONCLUSION: The sequence variation data of HPV18 may lay a foundation for the virus diagnosis, further study of cervical cancer and vaccine design in central China.
Subject(s)
Genetic Variation , Human papillomavirus 18 , Oncogene Proteins, Viral , Papillomavirus E7 Proteins , Phylogeny , Oncogene Proteins, Viral/genetics , China , Humans , Human papillomavirus 18/genetics , Human papillomavirus 18/classification , Papillomavirus E7 Proteins/genetics , Capsid Proteins/genetics , Female , Epitopes, T-Lymphocyte/genetics , Papillomavirus Infections/virology , Repressor Proteins/genetics , Epitopes, B-Lymphocyte/genetics , DNA-Binding ProteinsABSTRACT
Three-stranded DNA-RNA structures known as R-loops that form during papillomavirus transcription can cause transcription-replication conflicts and lead to DNA damage. We found that R-loops accumulated at the viral early promoter in human papillomavirus (HPV) episomal cells but were greatly reduced in cells with integrated HPV genomes. RNA-DNA helicases unwind R-loops and allow for transcription and replication to proceed. Depletion of the RNA-DNA helicase senataxin (SETX) using siRNAs increased the presence of R-loops at the viral early promoter in HPV-31 (CIN612) and HPV-16 (W12) episomal HPV cell lines. Depletion of SETX reduced viral transcripts in episomal HPV cell lines. The viral E2 protein, which binds with high affinity to specific palindromes near the promoter and origin, complexes with SETX, and both SETX and E2 are present at the viral p97 promoter in CIN612 and W12 cells. SETX overexpression increased E2 transcription activity on the p97 promoter. SETX depletion also significantly increased integration of viral genomes in CIN612 cells. Our results demonstrate that SETX resolves viral R-loops to proceed with HPV transcription and prevent genome integration.IMPORTANCEPapillomaviruses contain small circular genomes of approximately 8 kilobase pairs and undergo unidirectional transcription from the sense strand of the viral genome. Co-transcriptional R-loops were recently reported to be present at high levels in cells that maintain episomal HPV and were also detected at the early viral promoter. R-loops can inhibit transcription and DNA replication. The process that removes R-loops from the PV genome and the requisite enzymes are unknown. We propose a model in which the host RNA-DNA helicase senataxin assembles on the HPV genome to resolve R-loops in order to maintain the episomal status of the viral genome.
Subject(s)
DNA Helicases , Multifunctional Enzymes , Promoter Regions, Genetic , R-Loop Structures , RNA Helicases , Humans , RNA Helicases/genetics , RNA Helicases/metabolism , Multifunctional Enzymes/genetics , Multifunctional Enzymes/metabolism , DNA Helicases/metabolism , DNA Helicases/genetics , R-Loop Structures/genetics , Plasmids/genetics , Virus Replication , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Papillomaviridae/genetics , Genome, Viral , Oncogene Proteins, Viral/genetics , Oncogene Proteins, Viral/metabolism , Transcription, Genetic , Cell Line , DNA, Viral/geneticsABSTRACT
Human papillomavirus (HPV) type 81 has recently become one of the most common low-risk HPV types; however, literature focusing on it is limited. This study aimed to analyze the reasons for the increased detection rate of HPV81 and investigate its evolving pathogenicity. We analyzed the detection rates and trends of HPV81 in 229 061 exfoliated cervical cell samples collected from 2014 to 2023; collected samples of HPV81 single infections from two different time periods; and analyzed the allele frequencies, positive selection, viral load, persistent infection capacity, and pathogenicity of E6 and E7 genotypes. We found that the detection rate of HPV81 ranked first among the low-risk types in exfoliated cervical cells and exhibited a significantly increasing trend (p < 0.001). The frequency of the E6 prototype allele of HPV81 (n = 317) was significantly increased (p = 0.018) and demonstrated the strongest adaptive capacity. The viral load and persistent infection capacity of the E6 prototype were significantly higher than those of the mutants, thus serving as key drivers for increasing the detection rate of HPV81 and enhancing its pathogenicity. The viral load was positively correlated with persistent infection capacity and pathogenicity. Persistent infection was a crucial factor in the pathogenicity of HPV81. Successful adaptive evolution of HPV81 is accompanied by enhanced pathogenicity.
Subject(s)
Genotype , Papillomavirus Infections , Persistent Infection , Polymorphism, Genetic , Viral Load , Humans , Papillomavirus Infections/virology , Female , Persistent Infection/virology , Cervix Uteri/virology , Cervix Uteri/pathology , Adult , Papillomaviridae/genetics , Papillomaviridae/pathogenicity , Papillomaviridae/classification , Papillomaviridae/isolation & purification , Gene Frequency , Oncogene Proteins, Viral/genetics , Virulence/genetics , Alphapapillomavirus/genetics , Alphapapillomavirus/pathogenicity , Alphapapillomavirus/classification , Alphapapillomavirus/isolation & purification , Human Papillomavirus VirusesABSTRACT
Human papillomavirus (HPV), a common cause of sexually transmitted diseases, may cause warts and lead to various types of cancers, which makes it important to understand the risk factors associated with it. HPV is the leading risk factor and plays a crucial role in the progression of cervical cancer. Viral oncoproteins E6 and E7 play a pivotal role in this process. Beyond cervical cancer, HPV-associated cancers of the mouth and throat are also increasing. HPV can also contribute to other malignancies like penile, vulvar, and vaginal cancers. Emerging evidence links HPV to these cancers. Research on the oncogenic effect of HPV is still ongoing and explorations of screening techniques, vaccination, immunotherapy and targeted therapeutics are all in progress. The present review offers valuable insight into the current understanding of the role of HPV in cancer and its potential implications for treatment and prevention in the future.
Subject(s)
Papillomavirus Infections , Uterine Cervical Neoplasms , Humans , Papillomavirus Infections/virology , Papillomavirus Infections/complications , Female , Uterine Cervical Neoplasms/virology , Uterine Cervical Neoplasms/etiology , Papillomaviridae/pathogenicity , Neoplasms/virology , Neoplasms/therapy , Oncogene Proteins, Viral/metabolism , Oncogene Proteins, Viral/genetics , Risk Factors , MaleABSTRACT
Despite epidermal turnover, the skin is host to a complex array of microbes, including viruses, such as HPV, which must infect and manipulate skin keratinocyte stem cells (KSCs) to survive. This crosstalk between the virome and KSC populations remains largely unknown. Here, we investigated the effect of HPV8 on KSCs using various mouse models. We observed that the HPV8 early region gene E6 specifically caused Lrig1+ hair follicle junctional zone KSC proliferation and expansion, which would facilitate viral transmission. Within Lrig1+ KSCs specifically, HPV8 E6 bound intracellular p300 to phosphorylate the STAT3 transcriptional regulatory node. This induced ΔNp63 expression, resulting in KSC expansion into the overlying epidermis. HPV8 was associated with 70% of human actinic keratoses. Together, these results define the "hit-and-run" mechanism for HPV8 in human actinic keratosis as an expansion of KSCs, which lack melanosome protection and are thus susceptible to sun light-induced malignant transformation.
Subject(s)
Cell Proliferation , Keratinocytes , Keratosis, Actinic , Oncogene Proteins, Viral , Papillomavirus Infections , STAT3 Transcription Factor , Stem Cells , STAT3 Transcription Factor/metabolism , Keratinocytes/virology , Keratinocytes/metabolism , Keratinocytes/pathology , Humans , Keratosis, Actinic/pathology , Keratosis, Actinic/metabolism , Keratosis, Actinic/virology , Animals , Mice , Stem Cells/metabolism , Stem Cells/virology , Oncogene Proteins, Viral/metabolism , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/virology , Papillomavirus Infections/pathology , Papillomavirus Infections/metabolism , Papillomavirus Infections/complications , Disease Models, Animal , FemaleABSTRACT
During entry, human papillomavirus (HPV) traffics from the endosome to the trans Golgi network (TGN) and Golgi and then the nucleus to cause infection. Although dynein is thought to play a role in HPV infection, how this host motor recruits the virus to support infection and which entry step(s) requires dynein are unclear. Here we show that the dynein cargo adaptor BICD2 binds to the HPV L2 capsid protein during entry, recruiting HPV to dynein for transport of the virus along the endosome-TGN/Golgi axis to promote infection. In the absence of BICD2 function, HPV accumulates in the endosome and TGN and infection is inhibited. Cell-based and in vitro binding studies identified a short segment near the C-terminus of L2 that can directly interact with BICD2. Our results reveal the molecular basis by which the dynein motor captures HPV to promote infection and identify this virus as a novel cargo of the BICD2 dynein adaptor.
Subject(s)
Capsid Proteins , Human papillomavirus 16 , Oncogene Proteins, Viral , Papillomavirus Infections , Humans , Capsid Proteins/metabolism , Human papillomavirus 16/metabolism , Oncogene Proteins, Viral/metabolism , Papillomavirus Infections/metabolism , Papillomavirus Infections/virology , Dyneins/metabolism , Endosomes/metabolism , Endosomes/virology , trans-Golgi Network/metabolism , trans-Golgi Network/virology , Virus Internalization , Protein Binding , HeLa Cells , Microtubule-Associated Proteins/metabolism , Cytoplasmic Dyneins/metabolismABSTRACT
Integration of the human papillomavirus (HPV) genome into the cellular genome is a key event that leads to constitutive expression of viral oncoprotein E6/E7 and drives the progression of cervical cancer. However, HPV integration patterns differ on a case-by-case basis among related malignancies. Next-generation sequencing technologies still face challenges for interrogating HPV integration sites. In this study, utilizing Nanopore long-read sequencing, we identified 452 and 108 potential integration sites from the cervical cancer cell lines (CaSki and HeLa) and five tissue samples, respectively. Based on long Nanopore chimeric reads, we were able to analyze the methylation status of the HPV long control region (LCR), which controls oncogene E6/E7 expression, and to identify transcriptionally-active integrants among the numerous integrants. As a proof of concept, we identified an active HPV integrant in between RUNX2 and CLIC5 on chromosome 6 in the CaSki cell line, which was supported by ATAC-seq, H3K27Ac ChIP-seq, and RNA-seq analysis. Knockout of the active HPV integrant, by the CRISPR/Cas9 system, dramatically crippled cell proliferation and induced cell senescence. In conclusion, identifying transcriptionally-active HPV integrants with Nanopore sequencing can provide viable targets for gene therapy against HPV-associated cancers.
Subject(s)
Genetic Therapy , Nanopore Sequencing , Papillomavirus Infections , Uterine Cervical Neoplasms , Virus Integration , Humans , Uterine Cervical Neoplasms/virology , Female , Nanopore Sequencing/methods , Virus Integration/genetics , Genetic Therapy/methods , Papillomavirus Infections/virology , Cell Line, Tumor , HeLa Cells , Oncogene Proteins, Viral/genetics , High-Throughput Nucleotide Sequencing/methods , Papillomaviridae/genetics , Human Papillomavirus VirusesABSTRACT
The nonsense-mediated mRNA decay (NMD) pathway and the p53 pathway, linked to tumorgenesis, are also promising targets for cancer treatment. NMD plays an important role in RNA quality control, while the p53 pathway is involved in cancer suppression. However, their individual and combined effects on cervical cancer are poorly understood. In this study, we evaluated the impacts of NMD inhibitor, Mouse double minute 2 homolog (MDM2) inhibitor, and their combination on cell apoptosis, cell cycle, and p53 target genes in human papillomavirus-18-positive HeLa cells. Our findings revealed that XR-2 failed to activate p53 or induce apoptosis in HeLa cells, whereas SMG1 (serine/threonine-protein kinase 1) inhibitor repressed cell proliferation at high concentrations. Notably, the combination of these 2 agents significantly inhibited cell proliferation, arrested the cell cycle, and triggered cell apoptosis. Mechanistically, MDM2 inhibitor and NMD inhibitor likely exert a synergistically through the truncated E6 protein. These results underscore the potential of employing a combination of MDM2 inhibitor and NMD inhibitor as a promising candidate for the clinical treatment of human papillomavirus-infected tumors.
Subject(s)
Apoptosis , Cell Proliferation , Proto-Oncogene Proteins c-mdm2 , Humans , Proto-Oncogene Proteins c-mdm2/metabolism , Proto-Oncogene Proteins c-mdm2/antagonists & inhibitors , Apoptosis/drug effects , HeLa Cells , Cell Proliferation/drug effects , Nonsense Mediated mRNA Decay/drug effects , Tumor Suppressor Protein p53/metabolism , Drug Synergism , Oncogene Proteins, Viral/metabolism , Uterine Cervical Neoplasms/drug therapy , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/metabolism , Repressor Proteins/metabolism , Repressor Proteins/antagonists & inhibitors , Repressor Proteins/genetics , DNA-Binding ProteinsABSTRACT
Oncogenic HPV E6 proteins have a PDZ-binding motif (PBM) which plays important roles in both the viral life cycle and tumor development. The PBM confers interaction with a large number of different PDZ domain-containing substrates, one of which is Sorting Nexin 27. This protein is part of the retromer complex and plays an important role in endocytic sorting pathways. It has been shown that at least two SNX27 interacting partners, GLUT1 and TANC2, are aberrantly trafficked due to the E6 PBM-dependent interaction with SNX27. To investigate further which other components of the endocytic trafficking pathway might be affected by the SNX27-HPV E6 interaction, we analyzed the SNX27 proteome interaction profile in a previously described HeLa cell line expressing GFP-SNX27, both in the presence and absence of the HPV-18 E6 oncoprotein. In this study, we identify a novel interacting partner of SNX27, secreted glycoprotein EMILIN2, whose release is blocked by HPV18 E6 in a PBM-dependent manner. Mechanistically, E6 can block EMILIN2 interaction with the WNT1 ligand, thereby enhancing WNT1 signaling and promoting cell proliferation. IMPORTANCE: This study demonstrates that HPV E6 blocks EMILIN2 inhibition of WNT1 signaling, thereby enhancing cell proliferation in HPV-positive tumor cells. This involves a novel mechanism whereby the E6 PBM actually contributes toward enhancing the interaction between SNX27 and EMILIN2, suggesting that the mode of recognition of SNX27 by E6 and EMILIN2 is different. This is the first example of the E6 PBM altering a PDZ domain-containing protein to enhance potential substrate recognition.
Subject(s)
Human papillomavirus 18 , Oncogene Proteins, Viral , Sorting Nexins , Wnt Signaling Pathway , Humans , DNA-Binding Proteins , HEK293 Cells , HeLa Cells , Human papillomavirus 18/metabolism , Human papillomavirus 18/genetics , Oncogene Proteins, Viral/metabolism , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/virology , Papillomavirus Infections/metabolism , PDZ Domains , Protein Binding , Repressor Proteins/metabolism , Repressor Proteins/genetics , Sorting Nexins/metabolism , Sorting Nexins/geneticsABSTRACT
BACKGROUND/AIM: Human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) is clinically and immunologically distinct from HPV-negative HNSCC. Herein, we investigated the presence of tumor antigens HPV E6/E7 and wild-type p53-specific T-cell responses, and the impact of immune checkpoint blockade in patients with HPV-positive HNSCC. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) from patients with HPV-positive HNSCC were stimulated with HPV E6/E7 or wild-type p53-derived peptide mixture and evaluated using the interferon-γ enzyme-linked immunosorbent spot assay. Flow cytometry was performed to analyze the proportion of T-cell subsets and T cells expressing immune checkpoint molecules. RESULTS: HPV E6/E7-specific T cells were detected in 22 (95.7%) of 23 patients, whereas wild-type p53-specific T cells were detected in 3 (15.0%) of 20 patients. Seven (43.8%) of 16 patients exhibited wild-type p53-specific T-cell responses, as determined using whole proteins instead of peptides. Immune checkpoint blockade enhanced wild-type p53-specific T-cell responses in 9 (45.0%) of 20 patients. Flow cytometric analysis of PBMCs revealed that responders exhibiting enhanced wild-type p53-specific T-cell responses following immune checkpoint blockade had a significantly higher proportion of Ki-67+CD4+ T cells, Ki-67+CD8+ T cells, regulatory T cells, PD-1+CD4+ T cells, and TIM-3+CD4+ T cells than non-responders. CONCLUSION: Our findings indicate that tumor antigen-specific T cells are present in the peripheral blood of patients with HPV-positive HNSCC. Blockade of checkpoint pathways can enhance T-cell responses in certain patients, probably via activated T cells, Tregs, and/or exhausted CD4+ T cells.
Subject(s)
Head and Neck Neoplasms , Immune Checkpoint Inhibitors , Papillomavirus Infections , Squamous Cell Carcinoma of Head and Neck , Humans , Immune Checkpoint Inhibitors/therapeutic use , Immune Checkpoint Inhibitors/pharmacology , Squamous Cell Carcinoma of Head and Neck/immunology , Squamous Cell Carcinoma of Head and Neck/virology , Squamous Cell Carcinoma of Head and Neck/drug therapy , Male , Female , Middle Aged , Aged , Head and Neck Neoplasms/immunology , Head and Neck Neoplasms/virology , Papillomavirus Infections/immunology , Papillomavirus Infections/virology , Antigens, Neoplasm/immunology , Oncogene Proteins, Viral/immunology , Tumor Suppressor Protein p53/immunology , Adult , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Papillomaviridae/immunology , T-Lymphocytes/immunology , Human Papillomavirus VirusesABSTRACT
Human papillomavirus (HPV) 16 and 18 infections are related to many human cancers. Despite several preventive vaccines for high-risk (hr) HPVs, there is still an urgent need to develop therapeutic HPV vaccines for targeting pre-existing hrHPV infections and lesions. In this study, we developed a lipid nanoparticle (LNP)-formulated mRNA-based HPV therapeutic vaccine (mHTV)-03E2, simultaneously targeting the E2/E6/E7 of both HPV16 and HPV18. mHTV-03E2 dramatically induced antigen-specific cellular immune responses, leading to significant CD8+ T cell infiltration and cytotoxicity in TC-1 tumors derived from primary lung epithelial cells of C57BL/6 mice expressing HPV E6/E7 antigens, mediated significant tumor regression, and prolonged animal survival, in a dose-dependent manner. We further demonstrated significant T cell immunity against HPV16/18 E6/E7 antigens for up to 4 months post-vaccination in immunological and distant tumor rechallenging experiments, suggesting robust memory T cell immunity against relapse. Finally, mHTV-03E2 synergized with immune checkpoint blockade to inhibit tumor growth and extend animal survival, indicating the potential in combination therapy. We conclude that mHTV-03E2 is an excellent candidate therapeutic mRNA vaccine for treating malignancies caused by HPV16 or HPV18 infections.
Subject(s)
Oncogene Proteins, Viral , Papillomavirus Infections , Papillomavirus Vaccines , RNA, Messenger , Animals , Mice , Papillomavirus Vaccines/immunology , Humans , Papillomavirus Infections/immunology , Papillomavirus Infections/virology , Papillomavirus Infections/therapy , Papillomavirus Infections/prevention & control , Female , Oncogene Proteins, Viral/immunology , Oncogene Proteins, Viral/genetics , RNA, Messenger/genetics , RNA, Messenger/immunology , Nanoparticles/chemistry , Human papillomavirus 16/immunology , Human papillomavirus 16/genetics , Mice, Inbred C57BL , Human papillomavirus 18/immunology , Human papillomavirus 18/genetics , Papillomavirus E7 Proteins/immunology , Papillomavirus E7 Proteins/genetics , Cancer Vaccines/immunology , Cancer Vaccines/administration & dosage , Cell Line, Tumor , Disease Models, Animal , CD8-Positive T-Lymphocytes/immunology , Repressor Proteins/immunology , Repressor Proteins/genetics , DNA-Binding Proteins , LiposomesABSTRACT
Cervical cancer (CC) and other malignant malignancies are acknowledged to be primarily caused by persistent human papillomavirus (HPV) infection. Historically, vaccinations against viruses that produce neutralizing antibodies unique to the virus have been an affordable way to manage viral diseases. CC risk is decreased, but not eliminated, by HPV vaccinations. Since vaccinations have been made available globally, almost 90% of HPV infections have been successfully avoided. On the lesions and diseases that are already present, however, no discernible treatment benefit has been shown. As a result, therapeutic vaccines that elicit immune responses mediated by cells are necessary for the treatment of established infections and cancers. mRNA vaccines possess remarkable potential in combating viral diseases and malignancy as a result of their superior industrial production, safety, and efficacy. Furthermore, considering the expeditiousness of production, the mRNA vaccine exhibits promise as a therapeutic approach targeting HPV. Given that the HPV-encoded early proteins, including oncoproteins E6 and E7, are consistently present in HPV-related cancers and pre-cancerous lesions and have crucial functions in the progression and persistence of HPV-related diseases, they serve as ideal targets for therapeutic HPV vaccines. The action mechanism of HPV and HPV-related cancer mRNA vaccines, their recent advancements in clinical trials, and the potential for their therapeutic applications are highlighted in this study, which also offers a quick summary of the present state of mRNA vaccines. Lastly, we highlight a few difficulties with mRNA HPV vaccination clinical practice and provide our thoughts on further advancements in this quickly changing sector. It is expected that mRNA vaccines will soon be produced quickly for clinical HPV prevention and treatment.