Human papillomavirus type 38 E6 and E7 act as tumour promoters during chemically induced skin carcinogenesis

Many findings support a possible involvement of a subgroup of human papillomaviruses (HPVs), called cutaneous beta HPV types, in the development of non-melanoma skin cancer. The skin of transgenic (Tg) mice expressing viral oncoproteins E6 and E7 from different cutaneous beta HPV types, including HPV38, showed an increased susceptibility to UV-induced and/or chemically induced skin carcinogenesis compared with wild-type animals. In this study, we show that beta HPV38 E6 and E7 oncoproteins act as promoter and progression factors in multi-stage skin carcinogenesis, strongly cooperating with the initiator and DNA damage agent 7,12-dimethylbenz[a]anthracene. In contrast, exposure of HPV38 E6/E7 Tg mice to the promoter 12-O-tetradecanoylphorbol-13-acetate did not significantly result in the development of skin lesions. These findings further support the role of beta HPV types in skin carcinogenesis, providing additional insight into their precise contribution to the multi-step process.

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Immune Enhancement of Skin Carcinogenesis by CD4+ T Cells

Journal of Experimental Medicine ◽

10.1084/jem.20021047 ◽

2003 ◽

Vol 197 (8) ◽

pp. 1017-1028 ◽

Cited By ~ 115

Author(s):

Dylan Daniel ◽

Nicole Meyer-Morse ◽

Emily K. Bergsland ◽

Kerstin Dehne ◽

Lisa M. Coussens ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Skin Lesions ◽

Skin Carcinogenesis ◽

Neoplastic Progression ◽

Multi Stage ◽

Malignant Lesions ◽

Immune Enhancement ◽

Metalloproteinase 9 ◽

Reduced Activity

In a transgenic model of multi-stage squamous carcinogenesis induced by human papillomavirus (HPV) oncogenes, infiltrating CD4+ T cells can be detected in both premalignant and malignant lesions. The lymph nodes that drain sites of epidermal neoplasia contain activated CD4+ T cells predominantly reactive toward Staphylococcal bacterial antigens. HPV16 mice deficient in CD4+ T cells were found to have delayed neoplastic progression and a lower incidence of tumors. This delay in carcinogenesis is marked by decreased infiltration of neutrophils, and reduced activity of matrix metalloproteinase-9, an important cofactor for tumor progression in this model. The data reveal an unexpected capability of CD4 T cells, whereby, proinflammatory CD4+ T cells, apparently responding to bacterial infection of dysplastic skin lesions, can inadvertently enhance neoplastic progression to invasive cancer.

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Cross-talk of cutaneous beta human papillomaviruses and the immune system: determinants of disease penetrance

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2018.0287 ◽

2019 ◽

Vol 374 (1773) ◽

pp. 20180287 ◽

Cited By ~ 6

Author(s):

Assunta Venuti ◽

Stefan Lohse ◽

Massimo Tommasino ◽

Sigrun Smola

Keyword(s):

Immune System ◽

Cross Talk ◽

Genetic Disorder ◽

Hpv Infection ◽

Human Papillomaviruses ◽

Skin Carcinogenesis ◽

Anogenital Region ◽

Disease Penetrance ◽

Hpv Types ◽

Mucosal Hpvs

Human papillomaviruses (HPVs) infect the epithelia of skin or mucosa, where they can induce hyperproliferative lesions. More than 220 different HPV types have been characterized and classified into five different genera. Mucosal high-risk HPVs are causative for cancers of the anogenital region and oropharynx. Clinical data from patients with the rare genetic disorder epidermodysplasia verruciformis (EV) indicate that genus beta-HPVs cooperate with ultraviolet (UV) radiation in the development of cutaneous squamous cell carcinoma. In addition, epidemiological and biological findings indicate that beta-HPV types play a role in UV-mediated skin carcinogenesis also in non-EV individuals. However, the mechanisms used by these cutaneous viruses to promote epithelial carcinogenesis differ significantly from those of mucosal HPVs. Recent studies point to a delicate cross-talk of beta-HPVs with the cell-autonomous immunity of the host keratinocytes and the local immune microenvironment that eventually determines the fate of cutaneous HPV infection and the penetrance of disease. This review gives an overview of the critical interactions of genus beta-HPVs with the local immune system that allow the virus to complete its life cycle, to escape from extrinsic immunity, and eventually to cause chronic inflammation contributing to skin carcinogenesis. This article is part of the theme issue ‘Silent cancer agents: multi-disciplinary modelling of human DNA oncoviruses’.

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A broad range of human papillomavirus types detected with a general PCR method suitable for analysis of cutaneous tumours and normal skin

Journal of General Virology ◽

10.1099/0022-1317-80-9-2437 ◽

1999 ◽

Vol 80 (9) ◽

pp. 2437-2443 ◽

Cited By ~ 336

Author(s):

Ola Forslund ◽

Annika Antonsson ◽

Peter Nordin ◽

Bo Stenquist ◽

Bengt Göran Hansson

Keyword(s):

Sequence Similarity ◽

Normal Skin ◽

Pcr Primers ◽

Skin Surface ◽

Skin Lesions ◽

Human Papillomaviruses ◽

Reading Frame ◽

Skin Biopsies ◽

Pcr Method ◽

Hpv Types

A pair of degenerate PCR primers (FAP59/64) was designed from two relatively conserved regions of the L1 open reading frame of most human papillomaviruses (HPV). The size of the generated amplicon was about 480 bp. PCR using these primers was found capable of amplifying DNA from 87% (65/75) of the HPV types tested, its sensitivity being 1–10 copies for HPV-5, -20 and -30 clones. HPV was found in 63% (5/8) of tumour samples and in 63% (5/8) of normal skin biopsies from patients with various cutaneous tumours. HPV-5, HPV-8, HPV-12, HPVvs20-4 and six putatively novel HPV types were identified. No correlation was found to exist between specific HPV and tumour types. Skin surface swab samples from one or more sites on three of four healthy volunteers were found to contain HPV, types 12 and 49 being identified, as well as eight novel HPV types, two of which were also found among the patients. In all, HPV was detected in 75% (9/12) of those tested, five HPV types and 12 novel candidate types being identified, and 37% (7/19) of HPV-positive samples were found to manifest more than one HPV type. All the HPV detected manifested high degrees of nucleotide sequence similarity with HPV types associated with skin lesions and epidermodysplasia verruciformis. The overall HPV finding in the skin samples was 50% (20/40) using the FAP primers as compared to 18% (7/40) using another PCR test designed for skin types. The results thus suggest the new method to be sensitive and generally applicable for detecting cutaneous HPV.

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Pseudovirion-binding and neutralizing antibodies to cutaneous human papillomaviruses (HPV) correlated with the presence of HPV DNA in skin

Journal of General Virology ◽

10.1099/vir.0.048561-0 ◽

2013 ◽

Vol 94 (5) ◽

pp. 1096-1103 ◽

Cited By ~ 5

Author(s):

Helena Faust ◽

Kristin Andersson ◽

Ola Forslund ◽

Joakim Dillner

Keyword(s):

Antibody Response ◽

Cell Carcinoma ◽

Neutralizing Antibodies ◽

Skin Lesions ◽

Human Papillomaviruses ◽

Serum Samples ◽

Specific Antibody Response ◽

Hpv Dna ◽

Immunosuppressed Patients ◽

Hpv Types

Whereas the antibody response to the anogenital human papillomaviruses (HPVs) is known to be mainly type-specific, correlated with the presence of viral DNA and mainly directed to conformational epitopes of the virion, it is not known if this applies also to the antibody response to cutaneous HPVs. For 434 non-immunosuppressed patients with skin lesions (squamous cell carcinoma and basal cell carcinoma of the skin, actinic keratosis and benign skin lesions), we compared HPV DNA status with seroreactivity to HPV pseudovirions (PsV) and to GST-L1 fusion proteins from HPV types -5, -6, -15, -16, -32 and -38. Biopsies from the skin lesions were tested for the presence of HPV DNA using three different PCR methods, with typing by sequencing. Serum samples from subjects with HPV DNA-positive biopsies and randomly selected serum samples from subjects with HPV DNA-negative biopsies were also tested with neutralization assays with HPV5, -38 and -76 PsV. Agreement of the three serological methods varied from poor to moderate. Type-specific seroprevalences among patients positive for the same type of HPV DNA (sensitivity of serology) was improved with the PsV-based method (mean of 40 %, maximum 63 %) compared with the GST-L1 method (mean of 20 %, maximum of 25 %). Neutralization was the most sensitive assay for HPV38 (50 %). In summary, cutaneous HPVs also appear to induce a type-specific antibody response that correlates with the presence of HPV DNA and that can be detected with improved sensitivity using PsV-based serology.

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Transforming Properties of Beta-3 Human Papillomavirus E6 and E7 Proteins

mSphere ◽

10.1128/msphere.00398-20 ◽

2020 ◽

Vol 5 (4) ◽

Cited By ~ 1

Author(s):

Lucia Minoni ◽

Maria Carmen Romero-Medina ◽

Assunta Venuti ◽

Cécilia Sirand ◽

Alexis Robitaille ◽

...

Keyword(s):

High Risk ◽

Expression Profiles ◽

Expression Patterns ◽

Biological Properties ◽

Human Papillomaviruses ◽

Hpv E6 ◽

Hpv Types ◽

Beta 2 ◽

E6 And E7

ABSTRACT The beta human papillomaviruses (HPVs) are subdivided into 5 species (beta-1 to beta-5), and they were first identified in the skin. However, the beta-3 species appears to be more highly represented in the mucosal epithelia than in the skin. Functional studies have also highlighted that beta-3 HPV49 shares some functional similarities with mucosal high-risk (HR) HPV16. Here, we describe the characterization of the in vitro transforming properties of the entire beta-3 species, which includes three additional HPV types: HPV75, HPV76, and HPV115. HPV49, HPV75, and HPV76 E6 and E7 (E6/E7), but not HPV115 E6 and E7, efficiently inactivate the p53 and pRb pathways and immortalize or extend the life span of human foreskin keratinocytes (HFKs). As observed for HR HPV16, cell cycle deregulation mediated by beta-3 HPV E6/E7 expression leads to p16INK4a accumulation, whereas no p16INK4a was detected in beta-2 HPV38 E6/E7 HFKs. As shown for HPV49 E6, HPV75 and HPV76 E6s degrade p53 by an E6AP/proteasome-mediated mechanism. Comparative analysis of cellular gene expression patterns of HFKs containing E6 and E7 from HR HPV16, beta-3 HPV types, and beta-2 HPV38 further highlights the functional similarities of HR HPV16 and beta-3 HPV49, HPV75, and HPV76. The expression profiles of these four HPV HFKs show some similarities and diverge substantially from those of beta-3 HPV115 E6/E7 and beta-2 HPV38 E6/E7 HFKs. In summary, our data show that beta-3 HPV types share some mechanisms with HR HPV types and pave the way for additional studies aiming to evaluate their potential role in human pathologies. IMPORTANCE Human papillomaviruses are currently classified in different genera. Mucosal HPVs belonging to the alpha genus have been clearly associated with carcinogenesis of the mucosal epithelium at different sites. Beta HPV types have been classified as cutaneous. Although findings indicate that some beta HPVs from species 1 and 2 play a role, together with UV irradiation, in skin cancer, very little is known about the transforming properties of most of the beta HPVs. This report shows the transforming activity of E6 and E7 from beta-3 HPV types. Moreover, it highlights that beta-3 HPVs share some biological properties more extensively with mucosal high-risk HPV16 than with beta-2 HPV38. This report provides new paradigms for a better understanding of the biology of the different HPV types and their possible association with lesions at mucosal and/or cutaneous epithelia.

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Whole Genomic Analysis and Comparison of Two Canine Papillomavirus Type 9 Strains in Malignant and Benign Skin Lesions

Viruses ◽

10.3390/v12070736 ◽

2020 ◽

Vol 12 (7) ◽

pp. 736

Author(s):

Chia-Yu Chang ◽

Nanako Yamashita-Kawanishi ◽

Sonoka Tomizawa ◽

I-Li Liu ◽

Wei-Tao Chen ◽

...

Keyword(s):

Animal Species ◽

Reference Strain ◽

Genomic Analysis ◽

Skin Lesions ◽

Human Papillomaviruses ◽

Viral Oncoproteins ◽

E2 Protein ◽

Viral Plaque ◽

Malignant Lesions ◽

E6 And E7

Papillomaviruses (PVs) usually cause benign proliferative lesions in the stratified epithelium of various animal species. However, some high-risk types of PVs have been proven to lead to malignant transformations. In dogs, several canine papillomaviruses (CPVs) have been identified in malignant lesions and are suggested as one of the risk factors for the development of squamous cell carcinomas (SCCs). In the present study, the full genomes of two CPV9 strains from recurrent SCCs of Dog 1 and skin viral papilloma (viral plaque) of Dog 2 were sequenced. Alignment of the two CPV9 sequences with the genome of the reference CPV9 strain (accession no. JF800656.1) derived from a solitary pigmented plaque was performed. Compared with the reference strain, a 27 bp in-frame insertion in the E1 gene was identified in both CPV9 strains in this study. In comparison with the CPV9 strains derived from benign lesions, the CPV9 from the SCCs of Dog 1 exhibited a 328 bp deletion at the 3′ end of the E2 and spacer sequence, which encoded a truncated deduced E2 protein and a chimeric E8^E2 protein. However, there was no difference in the mRNA expression levels of viral oncoproteins of E6 and E7 between the two CPV9 cases, suggesting that the oncogenesis of CPV9 for malignant transformation might be different from that of human papillomaviruses. The roles of E2 and E8^E2 deleted CPV9 in the oncogenesis of benign and malignant lesions should be further investigated.

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Four novel human betapapillomaviruses of species 2 preferentially found in actinic keratosis

Journal of General Virology ◽

10.1099/vir.0.2008/001925-0 ◽

2008 ◽

Vol 89 (10) ◽

pp. 2467-2474 ◽

Cited By ~ 39

Author(s):

Nataša Vasiljević ◽

Kristina Hazard ◽

Joakim Dillner ◽

Ola Forslund

Keyword(s):

Cell Carcinoma ◽

Actinic Keratosis ◽

Skin Lesions ◽

Human Papillomaviruses ◽

Healthy Skin ◽

Viral Loads ◽

Seborrhoeic Keratosis ◽

Hpv Types ◽

Immunocompetent Patients

Recent studies have suggested an association between human papillomaviruses (HPVs), particularly species 2 members of the genus Betapapillomavirus, and squamous cell carcinoma (SCC) of the skin. As most of these viruses are uncharacterized, molecular characterization and epidemiology are needed to advance our understanding of their significance in carcinogenesis. This study determined the complete genomes of four betapapillomaviruses of species 2 from skin lesions designated HPV-107, -110 and -111 and FA75[KI88-03], an isolate of an unpublished HPV type, and analysed their prevalence and viral loads in biopsies from SCC, actinic keratosis (AK), basal cell carcinoma, seborrhoeic keratosis and the healthy skin of 263 immunocompetent patients by HPV type-specific real-time PCR assays. Seventeen patients (6.5 %) harboured at least one of the four HPV types in their lesion, whereas seven patients (2.7 %) harboured one or more of the HPV types in healthy skin. Overall, the four viruses were more common in AK than in healthy skin (odds ratio 5.0, 95 % confidence interval 1.4–17.5), but the prevalence and viral loads were low. This characterization of HPV-107, -110 and -111 and FA75[KI88-03] expands the heterogeneity of members of species 2 of the genus Betapapillomavirus. However, as these types were found in only a few samples and in low amounts, a possible role in carcinogenesis remains elusive.

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Genomic characterization of ten novel cutaneous human papillomaviruses from keratotic lesions of immunosuppressed patients

Journal of General Virology ◽

10.1099/vir.0.030593-0 ◽

2011 ◽

Vol 92 (7) ◽

pp. 1585-1594 ◽

Cited By ~ 35

Author(s):

Anja Köhler ◽

Marc Gottschling ◽

Kizzie Manning ◽

Mandy D. Lehmann ◽

Eric Schulz ◽

...

Keyword(s):

Phylogenetic Analyses ◽

Rolling Circle Amplification ◽

Organ Transplant ◽

Human Papillomaviruses ◽

Rolling Circle ◽

Non Melanoma Skin Cancer ◽

Immunosuppressed Patients ◽

Genomic Characterization ◽

Hpv Types

Viral warts from immunosuppressed organ transplant recipients (OTR) persist over years and may progress into non-melanoma skin cancer. The types of human papillomaviruses (HPV) in such lesions are different from that seen in the general population. A subset of these lesions is not infected with the classical wart-associated HPV types. In order to gain a better understanding of the HPV types in those lesions, we isolated ten novel HPVs from persisting keratotic lesions of immunosuppressed OTRs by rolling circle amplification and subsequent long-template PCR. Additionally, we sequenced and characterized the whole genome of the ten novel HPV types. Phylogenetic analyses revealed that nine HPV types belonged to the genus Gammapapillomavirus (γ-PV) and one to the genus Betapapillomavirus. In a phylogenetic analysis using L1 fragments of human and non-human PV types, primate papillomaviruses and our novel HPV types nested within the genus γ-PV in a highly polyphyletic pattern. This study significantly broadens the knowledge concerning the diversity and evolution of the poorly known γ-PV types.

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Phylogeny and polymorphism in the E6 and E7 of human papillomavirus: alpha-9 (HPV16, 31, 33, 52, 58), alpha-5 (HPV51), alpha-6 (HPV53, 66), alpha-7 (HPV18, 39, 59, 68) and alpha-10 (HPV6, 44) in women from Shanghai

Infectious Agents and Cancer ◽

10.1186/s13027-019-0250-9 ◽

2019 ◽

Vol 14 (1) ◽

Author(s):

Junwei Zhao ◽

Qin Zhan ◽

Junhan Guo ◽

Min Liu ◽

Yetian Ruan ◽

...

Keyword(s):

Phylogenetic Trees ◽

Intraepithelial Neoplasia ◽

Human Papillomaviruses ◽

Therapeutic Interventions ◽

Cervical Lesions ◽

Intraepithelial Lesion ◽

Species Groups ◽

Hpv Types ◽

E6 And E7 ◽

Alpha 7

Abstract Background Persistent infection with human papillomaviruses (HPVs) has been associated with cervical intraepithelial neoplasia (CIN) and cervical cancer. However, why only a fraction of HPV cases progress to cancer is still unclear. Methods We focused on the heterogeneity, classification, evolution and dispersal of variants for 14 common HPV types in 262 HPV-positive patients with cervical lesions. The E6 and E7 genes of HPV were sequenced and compared with the HPV reference for sequence analysis. Phylogenetic trees were constructed using the neighbour-joining tree method with MEGA 7.0. Results In this study, 233 E6 and 212 E7 sequences were successfully amplified by PCR, and these sequences were divided into 5 species groups: alpha-9 (HPV16, 31, 33, 52, 58), alpha-5 (HPV51), alpha-6 (HPV53, 66), alpha-7 (HPV18, 39, 59, 68) and alpha-10 (HPV6, 44). The incidence of high-grade squamous intraepithelial lesion (HSIL) in patients infected with alpha-9 HPV was significantly increased compared with other groups (P < 0.0001), especially HPV16 (P < 0.0001). Strikingly, E7 had significantly fewer nonsynonymous variants in the HSIL compared to <HSIL groups (P = 3.17× 10− 4). The A388C (K93 N) variation in HPV58 E6 can significantly reduce the risk of HSIL (P = 0.015). However, T7220G (D32E) variation in HPV16 E6 and A7689G (N29S) in HPV16 E7 increased the incidence of HSIL compared to the <HSIL group (P = 0.036 and 0.022). Conclusions Strict conservation of E7 is important for HPV carcinogenicity, especially N29 of HPV16. The findings in this work provide preventative/therapeutic interventions for HPV infections and CIN.

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UV Radiation Activates Toll-Like Receptor 9 Expression in Primary Human Keratinocytes, an Event Inhibited by Human Papillomavirus 38 E6 and E7 Oncoproteins

Journal of Virology ◽

10.1128/jvi.01123-17 ◽

2017 ◽

Vol 91 (19) ◽

Cited By ~ 7

Author(s):

Laura Pacini ◽

Maria Grazia Ceraolo ◽

Assunta Venuti ◽

Giusi Melita ◽

Uzma A. Hasan ◽

...

Keyword(s):

Transcription Factors ◽

Uv Radiation ◽

Uv Irradiation ◽

Human Keratinocytes ◽

Skin Carcinogenesis ◽

Primary Human Keratinocytes ◽

Tlr9 Expression ◽

Hpv Types ◽

E6 And E7 Oncoproteins ◽

E6 And E7

ABSTRACT Several lines of evidence indicate that cutaneous human papillomavirus (HPV) types belonging to the beta genus of the HPV phylogenetic tree synergize with UV radiation in the development of skin cancer. Accordingly, the E6 and E7 oncoproteins from some beta HPV types are able to deregulate pathways related to immune response and cellular transformation. Toll-like receptor 9 (TLR9), in addition to playing a role in innate immunity, has been shown to be involved in the cellular stress response. Using primary human keratinocytes as experimental models, we have shown that UV irradiation (and other cellular stresses) activates TLR9 expression. This event is closely linked to p53 activation. Silencing the expression of p53 or deleting its encoding gene affected the activation of TLR9 expression after UV irradiation. Using various strategies, we have also shown that the transcription factors p53 and c-Jun are recruited onto a specific region of the TLR9 promoter after UV irradiation. Importantly, the E6 and E7 oncoproteins from beta HPV38, by inducing the accumulation of the p53 antagonist ΔNp73α, prevent the UV-mediated recruitment of these transcription factors onto the TLR9 promoter, with subsequent impairment of TLR9 gene expression. This study provides new insight into the mechanism that mediates TLR9 upregulation in response to cellular stresses. In addition, we show that HPV38 E6 and E7 are able to interfere with this mechanism, providing another explanation for the possible cooperation of beta HPV types with UV radiation in skin carcinogenesis. IMPORTANCE Beta HPV types have been suggested to act as cofactors in UV-induced skin carcinogenesis by altering several cellular mechanisms activated by UV radiation. We show that the expression of TLR9, a sensor of damage-associated molecular patterns produced during cellular stress, is activated by UV radiation in primary human keratinocytes (PHKs). Two transcription factors known to be activated by UV radiation, p53 and c-Jun, play key roles in UV-activated TLR9 expression. The E6 and E7 oncoproteins from beta HPV38 strongly inhibit UV-activated TLR9 expression by preventing the recruitment of p53 and c-Jun to the TLR9 promoter. Our findings provide additional support for the role that beta HPV types play in skin carcinogenesis by preventing activation of specific pathways upon exposure of PHKs to UV radiation.

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