Equus caballus papillomavirus type 2 (EcPV2) in co-occurring vulvar and gastric lesions of a pony

Porcellato, Ilaria; Modesto, Paola; Cappelli, Katia; Varello, Katia; Peletto, Simone; Brachelente, Chiara; Martini, Isabella; Mechelli, Luca; Ferrari, Angelo; Ghelardi, Alessandro; Razzuoli, Elisabetta

doi:10.1016/j.rvsc.2020.06.003

Cited by 12 publications

(29 citation statements)

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“…In our study, the presence of EcPV2, which is the most common equine [40], was investigated by testing the samples for the DNA oncogene E6 positi 23 tumors, 21 (91%) resulted positive for EcPV2-E6 DNA. The lack of E6 posi two SCC samples could be due to DNA fragmentation, a frequent condition fixed and paraffin-embedded (FFPE) samples, although a specific primer pair duce an amplicon less than 100bp was used [29]. In humans, two pathways a in vulvar squamous cells carcinomas onset.…”

Section: Discussionmentioning

confidence: 99%

“…Two to three, 5 µm thick sections were cut from each FFPE block (both T and C samples) and subjected to DNA extraction for the evaluation of EcPV2 presence, which was carried out using the AllPrep DNA FFPE Kit (Qiagen, Venlo, the Netherlands) following the manufacturer's instructions. Previously described primers for EcPV2-E6 DNA and related specific probes were used for the virus detection [29], and beta-2-microglobulin (B2M) gene was targeted to assess DNA amplifiability. Primer sequences are reported in Table 1.…”

Section: Dna Extraction and Ecpv2 Detectionmentioning

confidence: 99%

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Equine Genital Squamous Cell Carcinoma Associated with EcPV2 Infection: RANKL Pathway Correlated to Inflammation and Wnt Signaling Activation

Mecocci

Porcellato

Armando

et al. 2021

Biology

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Equine genital squamous cell carcinomas (egSCCs) are among the most common equine tumors after sarcoids, severely impairing animal health and welfare. Equus caballus papillomavirus type 2 (EcPV2) infection is often related to these tumors. The aim of this study was to clarify the molecular mechanisms behind egSCCs associated with EcPV2 infection, investigating receptor activator of nuclear factor-kappa B ligand (RANKL) signaling in NF-kB pathway, together with the Wnt and IL17 signaling pathways. We analyzed the innate immune response through gene expression evaluation of key cytokines and transcription factors. Moreover, Ki67 index was assessed with immunohistochemistry. EcPV2-E6 DNA was checked, and viral presence was confirmed in 21 positive out to 23 cases (91%). Oncogene expression was confirmed in 14 cases (60.8%) for E6 and in 8 (34.7%) for E2. RANKL, nuclear factor kappa-light-chain-enhancer of activated B cells (NFKB)-p50, NFKBp65, interleukin (IL)-6, IL17, IL23p19, IL8, IL12p35, IL12p40, β-catenin (BCATN1), FOS like 1 (FOSL1), and lymphoid enhancer binding factor 1 (LEF1) showed a significant upregulation in tumor samples compared to healthy tissues. Our results describe an inflammatory environment characterized by the activation of RANKL/RANK and IL17 with the relative downstream pathways, and a positive modulation of inflammatory cytokines genes such as IL6 and IL8. Moreover, the increase of BCATN1, FOSL1, and LEF1 gene expression suggests an activation of both canonical and non-canonical Wnt signaling pathway that could be critical for carcinogenesis and tumor progression.

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Section: Discussionmentioning

confidence: 99%

Section: Dna Extraction and Ecpv2 Detectionmentioning

confidence: 99%

Equine Genital Squamous Cell Carcinoma Associated with EcPV2 Infection: RANKL Pathway Correlated to Inflammation and Wnt Signaling Activation

Mecocci

Porcellato

Armando

et al. 2021

Biology

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“…During extraction, samples were treated with DNase (RNase-Free DNase set, Qiagen). To evaluate DNA amplifiability, β-actin gene amplification was performed [7]; then, EcPV2-L1 DNA presence was tested using a Real-Time protocol previously described [7]. Briefly, TaqMan ® probe-based real-time PCR (CustomProbe 2× qPCR Master Mix, Canvax, Cordoba, Spain, cat.…”

Section: Detection Of Ecpv2 and Evaluation Of Oncogene Expressionmentioning

confidence: 99%

“…The fluorescence threshold limit was set automatically. Moreover, the expression of E2, E6, and L1 genes was evaluated by RT-real-time PCR using primers and probes (Table 2) and a protocol described by Porcellato and co-workers [7]. Briefly, RT was performed using a SuperScript™ IV VILO™ Master Mix (Invitrogen, ThermoFisher Scientific) according to the manufacturer's instructions.…”

Section: Detection Of Ecpv2 and Evaluation Of Oncogene Expressionmentioning

confidence: 99%

“…Particularly, h-HPV16 is the most common type (19.8%). Interestingly, equine papillomavirus type 2 (EcPV2) also has been recently associated with SCC of the penis, vulva, and stomach [7][8][9][10], as well as it has been recently detected in equine SCCL [11]. These data suggest that the horse is a good spontaneous animal model for studying the pathogenic mechanisms of this tumor in humans.…”

Section: Introductionmentioning

confidence: 99%

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Epithelial to Mesenchymal Transition (EMT) in a Laryngeal Squamous Cell Carcinoma of a Horse: Future Perspectives

Armando

Godizzi

Razzuoli³

et al. 2020

Animals

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Squamous cell carcinoma (SCC) is one of the most frequent tumors of skin and muco-cutaneous junctions in the horse. Equine papillomavirus type 2 (EcPV2) has been detected in equine SCC of the oral tract and genitals, and recently also in the larynx. As human squamous cell carcinoma of the larynx (SCCL), it is strongly etiologically associated with high-risk papillomavirus (h-HPV) infection. This study focuses on tumor cells behavior in a naturally occurring tumor that can undergo the so-called epithelial to mesenchymal transition (EMT). A SCCL in a horse was investigated by immunohistochemistry using antibodies against E-cadherin, pan-cytokeratin AE3/AE1, β-catenin, N-cadherin, vimentin, ZEB-1, TWIST, and HIF-1α. EcPV2 DNA detection and expression of oncogenes in SCC were investigated. A cadherin switch and an intermediate filaments rearrangement within primary site tumor cells together with the expression of the EMT-related transcription factors TWIST-1, ZEB-1, and HIF-1α were observed. DNA obtained from the tumor showed EcPV2 positivity, with E2 gene disruption and E6 gene dysregulation. The results suggest that equine SCCL might be a valuable model for studying EMT and the potential interactions between EcPV2 oncoproteins and the EMT process in SCCL.

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Equus caballus papillomavirus type 2 (EcPV2)‐associated benign penile lesions and squamous cell carcinomas

Tuomisto,

Virtanen,

Kegler

et al. 2024

Veterinary Medicine & Sci

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BackgroundSquamous cell carcinoma is the most common genital, ocular and gastric tumour in horses. Equus caballus papillomavirus type 2 (EcPV2) DNA has been detected in several studies in equine penile squamous cell carcinomas (SCCs) and precursor lesions providing evidence of a causal role of EcPV2 in equine genital SCCs. Recently, EcPV2 E6/E7 nucleic acids were also detected in equine gastric SCCs, but further studies are required to determine the role of EcPV2 infection in the pathogenesis of gastric SCC. EcPV2 nucleic acids have been rarely described in ocular SCCs and precursor lesions.ObjectivesTo investigate the presence of EcPV2 nucleic acids with polymerase chain reaction (PCR) and in situ hybridisation (ISH) in penile hyperplasias, papillomas and SCCs in horses and to determine whether EcPV2 nucleic acids can be detected in SCCs affecting other locations, including the stomach, ocular tissues and larynx.MethodsTwenty‐one archival formalin‐fixed paraffin embedded (FFPE) tissue samples, including 12 genital lesions comprising penile hyperplasias, papillomas and SCCs, 6 ocular SCCs, 2 gastric SCCs and 1 laryngeal SCC, were screened by PCR and ISH for EcPV2 E6/E7 DNA and mRNA. Archival FFPE tissue samples (eyelid and penile mucosa and preputium) from six horses without a diagnosis or history of neoplastic or papillomavirus‐associated disease were included as controls.ResultsEcPV2 nucleic acids were detected by PCR and ISH in all genital lesions (12/12) and gastric SCCs (2/2), in two ocular SCCs (2/6) and in one laryngeal SCC (1/1). In control horses, one eyelid sample was positive in PCR but not in ISH. The remaining control samples were negative for EcPV2 E6/E7 nucleic acids in PCR and ISH.ConclusionsThese results further support the role of EcPV2 infection in the development of equine genital SCCs and suggest that EcPV2 infection may also act as a predisposing factor for other SCCs in horses, including gastric, ocular and laryngeal SCCs.

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Equus caballus papillomavirus type 2 (EcPV2) in co-occurring vulvar and gastric lesions of a pony

Cited by 12 publications

References 20 publications

Equine Genital Squamous Cell Carcinoma Associated with EcPV2 Infection: RANKL Pathway Correlated to Inflammation and Wnt Signaling Activation

Equine Genital Squamous Cell Carcinoma Associated with EcPV2 Infection: RANKL Pathway Correlated to Inflammation and Wnt Signaling Activation

Epithelial to Mesenchymal Transition (EMT) in a Laryngeal Squamous Cell Carcinoma of a Horse: Future Perspectives

Equus caballus papillomavirus type 2 (EcPV2)‐associated benign penile lesions and squamous cell carcinomas

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