Acta Dermato-Venereologica 99-4CompleteContent | Page 29

462 SHORT COMMUNICATION High-risk Mucosal Human Papillomavirus Infection in Squamous Cell Carcinoma and Bowen’s Disease of the Hand Sonja DORFER 1 , Stefanie KANCZ 1 , Peter BIRNER 2 , Katharina STRASSER 1 , Reinhard KIRNBAUER 1 , Peter PETZELBAUER 1 , Sonja RADAKOVIC 1 , Lucie HARPAIN 1 , Hubert PEHAMBERGER 1,3 , Florian THALHAMMER 4 and Alessandra HANDISURYA 1 * Departments of 1 Dermatology, 2 Pathology and 4 Internal Medicine I, Medical University of Vienna, Waehringer Guertel 18-20, AT-1090 Vienna, and 3 Private Hospital Rudolfinerhaus, Vienna, Austria. *E-mail: [email protected] Accepted Jan 15, 2019; E-published Jan 17, 2019 Infections with mucosal high-risk (HR-) human pa- pillomaviruses (HPVs) cause mucosal cancers of the anogenital and oropharyngeal tract. Furthermore, they are associated with the development of skin cancers located on the hands. Available data on viral prevalence and type spectrum in these tumours are highly variable. Some studies show detection rates of mucosal HR-HPV in up to 90% (1–5), while others fail to detect viral DNA (6, 7). Viral presence is reported to be confined to tumours on peri- and subungual sites, or predominantly on proximal phalanges rather than ungual sites (2, 3). This study investigated cutaneous squamous cell carcino- mas (SCCs) and Bowen’s diseases (BDs), an SCC in situ, located on the hands for the presence of mucosal HR-HPV and the respective genotype(s). In addition, candidate bio- markers for viral activity (p53/p16/p21) and therapeutic in- tervention (epidermal growth factor receptor (EGFR)) were analysed and expression correlated to papillomaviral state. METHODS AND RESULTS The methodology is given in Appendix S1 1 . Mucosal HR-HPV DNA was detected in the majority (56.3%) of cutaneous SCCs (n =  9) and BDs (n = 28) located on the hands (Table SI 1 ). The percentage of HR-HPV-positive tumours was lower in SCCs (33.3%) compared to BDs (65.2%). Viral DNA was more frequently present in skin tumours of the nail unit (83.3%) and fing- ers (63.6%) than of the back of the hands (0%) or wrists (33.3%). The predominant genotype was HR-HPV16 (68.4%) followed by 33 (15.8%), 52 (10.5%) and 26 (5.3%). In the majority (94.4%) of virus-positive tumours, infection with a single virus type was iden- tified, in one SCC (5.6%) co-infection with HPV33/52. Viral E6/ E7 mRNA expression within HPV-positive, but not HPV-negative, tumours was confirmed by RNA in situ hybridization (Fig. 1A–D), indicating transcriptional activity of the viral oncogenes. Hybrid capture-2 results (Table SII 1 ) further corroborated the presence of mucosal HR-HPVs. Interestingly, viral loads in SCCs were lower (mean relative light units (RLUs) 2.7) than in BDs (mean RLUs 305.8); however, due to the small number of SCCs definite conclusions were impeded. To evaluate possible candidate biomar- kers for viral activity, expression of certain HR-HPV-dependent molecules was investigated. Most tumours (91.2%) did not display positive p53-immunostaining (Fig. 1E–H). Two BDs, lacking HR- HPV DNA, showed nuclear p53-immunostaining in the basal and lower suprabasal layers (Fig. 1H), while all others and all SCCs, irrespective of viral status, were p53-negative. In contrast, p16 expression was found in the majority (91.7%) of tumours, in 82.4% of virus-positive (Fig. 1I, J) and all virus- negative tumours (Fig. 1K, L). Both, staining intensities and https://www.medicaljournals.se/acta/content/abstract/10.2340/00015555-3115 1 doi: 10.2340/00015555-3115 Acta Derm Venereol 2019; 99: 462–463 Fig. 1. HPV16 E6/E7 mRNA was detected by RNA in situ hybridization in high-risk human papillomavirus (HR-HPV) DNA-positive squamous cell carcinomas (SCCs) (A) and Bowen’s diseases (BDs) (B), whereas HR- HPV DNA-negative SCCs (C) and BDs (D) lacked specific signals. Expression of p53 protein was restricted to a subset of virus-negative BDs (E–H). P16 expression was found in the vast majority of HR-HPV DNA-positive and -negative tumors (I–L). Nuclear immunostaining of p21 throughout the entire tumour was found uniformly in all specimens (M-P). Specific epidermal growth factor receptor (EGFR)-immunoreactivity was observed in 20.0% of HR-HPV DNA-positive and 28.6% of -negative tumours (Q–T). BDs were more frequently positive for EGFR compared with SCCs. numbers of p16-immunopositive cells, were similar between these 2 entities and observed differences were not significant. Only 8.3% of the samples, 1 SCC and 2 BDs, did not express p16 and these specimens were concomitantly HPV16 DNA-positive (not shown). Qualitative and quantitative expression of p16, regardless of viral presence, also did not differ between SCCs (88.9%) (Fig. 1I, K) and BDs (92.6%) (Fig. 1J, L). Immunostaining of p21 was found uniformly in 100% skin tumours. No differences of qualitative or quantitative p21 expres- sion were observed between HR-HPV DNA-positive (Fig. 1M, N) and -negative (Fig. 1O, P) tumours nor between SCCs (Fig. 1M, O) and BDs (Fig. 1N, P). Evaluation of EGFR showed positive immunostaining in 29.4% of the skin tumours; however, viral presence did not markedly af- This is an open access article under the CC BY-NC license. www.medicaljournals.se/acta Journal Compilation © 2019 Acta Dermato-Venereologica.