Basil S. Cherpelis

Case–Control Study of Cutaneous Human Papillomaviruses in Squamous Cell Carcinoma of the Skin

Background: Cutaneous human papillomavirus (HPV) infection may be a risk factor for squamous cell carcinoma (SCC) of the skin. Methods: To investigate the association between cutaneous HPV and SCC, a case–control study was conducted, including 173 SCC cases from a university dermatology clinic and 300 controls that screened negative for skin cancer. Serum antibodies against cutaneous HPV types in genera alpha, beta, gamma, mu, and nu were measured. Tumor tissue from 159 SCC cases was tested for the presence of DNA for genus-beta HPV types. Using logistic regression ORs and 95% confidence intervals (CI) were estimated for the associations between SCC and cutaneous HPV infection, adjusting for age and sex. The Bonferroni method was used to account for multiple comparisons. Results: SCC was positively associated with seropositivity to any genus-beta HPV type (OR, 1.93; 95% CI, 1.23–3.02), particularly with types in species-1 (OR, 1.86; 95% CI, 1.22–2.85). Type-specific associations with SCC were observed for HPV 8 (OR, 1.80; 95% CI, 1.14–2.84), 17 (OR, 1.59; 95% CI, 1.02–2.49) and HPV 10 from genus-alpha (OR, 2.24; 95% CI, 1.04–4.85). None of the type-specific associations remained statistically significant after correction for multiple comparisons. When DNA-positive SCC cases were compared with controls, strong serologic associations were observed for HPVs 5 (OR, 3.48; 95% CI, 1.27–9.59), 17 (OR, 3.36; 95% CI, 1.29–8.72), and 24 (OR, 3.79; 95% CI, 1.24–11.5). Conclusion: Genus-beta HPV infections were associated with SCC in our study population. Impact: Identifying the role of cutaneous HPV infection in SCC may lead to improved characterization of high-risk individuals and the development of novel prevention strategies. Cancer Epidemiol Biomarkers Prev; 21(8); 1303–13. ©2012 AACR.

Case–control Study of Merkel Cell Polyomavirus Infection and Cutaneous Squamous Cell Carcinoma

Background: Merkel cell polyomavirus (MCV) DNA has been reported in 0% to 25% of squamous cell carcinomas (SCC) occurring in immunocompetent individuals. We conducted the first serologic case–control study of MCV and SCC. Methods: Patients with histologically confirmed cutaneous SCC (n = 173) were recruited from a university dermatology clinic. Controls were individuals who screened negative for and had no history of skin or other cancers (n = 300). Levels of antibodies against capsid antigens for MCV and another polyomavirus, JC virus (JCV), were determined by fluorescent bead-based multiplex serology. Fresh-frozen tumor tissues were obtained from 145 SCC cases and tested for MCV DNA by multiplexed PCR. Associations between MCV seroreactivity and SCC were estimated by ORs and 95% CIs calculated using logistic regression with adjustment for age and sex. Results: MCV DNA was detected in SCC tumor tissues from 55 (38%) of 145 cases. A statistically significant association was observed between MCV seropositivity and MCV DNA-positive SCC (OR = 2.49, 95% CI = 1.03–6.04), with an almost four-fold association observed when comparing those with MCV antibodies in the fourth versus first quartiles (OR = 3.93, 95% CI = 1.43–10.76, Ptrend = 0.01). No significant associations were observed between MCV seropositivity and MCV DNA-negative SCC (OR = 1.38, 95% CI = 0.76–2.48) or between JCV seropositivity and MCV DNA-positive or DNA-negative SCC. Conclusion: Past exposure to MCV may be a risk factor for SCC. Impact: Understanding the role of viral infections in the development of nonmelanoma skin cancer could lead to novel prevention strategies. Cancer Epidemiol Biomarkers Prev; 21(1); 74–81. ©2011 AACR.

Case–control study of genus-beta human papillomaviruses in plucked eyebrow hairs and cutaneous squamous cell carcinoma

Cutaneous human papillomaviruses (HPV) have been reported in cutaneous squamous cell carcinoma (SCC). We conducted a clinic‐based case–control study to investigate the association between genus‐beta HPV DNA in eyebrow hairs (EBH) and SCC. EBH from 168 SCC cases and 290 controls were genotyped for genus‐beta HPV DNA. SCC tumors from a subset of cases (n = 142) were also genotyped. Viral load was determined in a subset of specimens positive for a single HPV type. Associations with SCC were estimated by odds ratios (OR) and 95% confidence intervals (CI) adjusted for age and sex using logistic regression. Statistical tests were two‐sided. EBH DNA prevalence was greater in cases (87%) than controls (73%) (p < 0.05), and the association with SCC increased with the number of HPV types present, (≥4 types vs. HPV‐negative: OR = 2.02, 95% CI = 1.07–3.80; ptrend = 0.02). Type‐specific associations were observed between SCC and DNA in EBH for HPV23 (OR = 1.90, 95% CI = 1.10–3.30) and HPV38 (OR = 1.84, 95% CI = 1.04–3.24). Additionally, when compared with the controls, the DNA prevalence in EBH was significantly higher among cases for 11 of the 25 genus‐beta types tested, when accounting for DNA for the same HPV type in the tumor (ORs = 3.44–76.50). Compared to controls, the mean viral DNA load in EBH among the selected cases was greater for HPV5, HPV8 and HPV24, but lower for HPV38. SCC cases were more likely than controls to have HPV DNA+ EBH for single and multiple HPV types, providing additional support for the potential role of genus‐beta HPV infections in SCC development.

Telomere length and risk of melanoma, squamous cell carcinoma, and basal cell carcinoma.

BACKGROUND Telomeres help maintain chromosomal structure and may influence tumorigenesis. We examined the association between telomere length and skin cancer in a clinic-based case-control study of 198 melanoma cases, 136 squamous cell carcinoma (SCC) cases, 185 basal cell carcinoma (BCC) cases, and 372 healthy controls. METHODS Cases were histologically confirmed patients treated at the Moffitt Cancer Center and University of South Florida Dermatology Clinic in Tampa, FL. Controls self-reported no history of cancer and underwent a skin cancer screening exam at study enrollment to rule out the presence of skin cancer. Quantitative real time PCR was used to measure telomere length in peripheral blood samples. RESULTS Melanoma patients had longer telomeres than controls (odds ratio (OR)=3.75; 95% confidence interval (CI): 2.02-6.94 for highest versus lowest tertile) (P for trend=<0.0001). In contrast, longer telomere length was significantly inversely associated with SCC (OR=0.01; 95% CI: 0.00-0.05 for highest versus lowest tertile) (P for trend=<0.0001) and BCC (OR=0.10; 95% CI: 0.06-0.19 for highest versus lowest tertile) (P for trend=<0.0001). CONCLUSION Telomere length may be involved in the development of skin cancer, although the effect on cancer risk differs for melanoma and non-melanoma carcinomas. Our findings suggest that long telomere length is positively associated with melanoma while inversely associated with SCC and BCC.

Case-control study of cutaneous human papillomavirus infection in Basal cell carcinoma of the skin.

Genus-β human papillomavirus (HPV) DNA has been detected in basal cell carcinoma (BCC) tumors, but most epidemiologic studies have not observed associations between genus-β HPV seropositivity and BCC. A clinic-based case-control study was conducted to investigate cutaneous HPV infection in BCC. BCC cases (n=224) were recruited from a dermatology clinic, and controls (n=300) were patients who were screened negative for skin cancer. Antibodies against cutaneous HPV types in genera α, β, γ, mu, and nu were measured, and tumors from a subset of BCC cases (n=195) were tested for HPV DNA. Overall associations were observed between BCC and seropositivity for HPV types in genus-α (odds ratio (OR)=1.61; 95% confidence interval (CI)=1.11-2.35), γ (OR=1.78; 95% CI=1.22-2.60), and mu (OR=1.56; 95% CI=1.06-2.30). BCC cases with β-HPV DNA in their tumors were more likely to be β-HPV seropositive than controls (OR=1.76; 95% CI=1.03-3.01), with type-specific associations observed for HPV8 and HPV23, whereas no association was observed between β-HPV seropositivity and β-HPV DNA-negative BCC. No concordance between seropositivity and tumor DNA status was observed for HPV types in genera α and γ. In conclusion, the combined serology and tumor DNA results suggest that β HPV types may have a role in BCC. Additional studies of BCC that assess HPV types in multiple genera are needed.

Comparison of MART-1 Frozen Sections to Permanent Sections Using a Rapid 19-Minute Protocol

BACKGROUND The use of melanoma‐associated antigen recognized by T cells (MART‐1) immunostain has been proposed as a useful adjunct to overcome the inherent difficulties in the use of frozen sections during Mohs surgery for the treatment of melanoma, but no studies have compared MART‐1 frozen sections with MART‐1 permanent sections. Current MART‐1 1‐hour protocols add significant time to the procedure. OBJECTIVE To determine whether there is a significant difference between frozen and permanent MART‐1 immunostained sections using a rapid 19‐minute protocol. METHODS Frozen and permanent sections stained with MART‐1 were made from dog‐ears excised during 25 reconstructions. A rapid 19‐minute protocol was used to stain the frozen tissue. The sections were examined blinded, and statistical analysis was performed to analyze the data. RESULTS No significant difference was found in number of keratinocytes, nuclear diameter of keratinocytes, number of melanocytes, melanocytic nuclear diameter, confluence, pagetoid spread, melanocytic nesting, or atypical melanocytes. CONCLUSIONS The 19‐minute protocol is a rapid and effective MART‐1 immunostain. Frozen sections stained with MART‐1 provide information equivalent to that obtained from MART‐1 stained permanent sections. Mohs surgeons using MART‐1 can feel confident that they have the same information as they would have obtained using permanent sections using the slow Mohs method. The authors have indicated no significant interest with commercial supporters.

Patterns and timing of sunlight exposure and risk of basal cell and squamous cell carcinomas of the skin - a case-control study

BackgroundNon-melanoma skin cancer (NMSC), comprised of basal (BCC) and squamous (SCC) cell carcinomas, is the most common cancer in Caucasians. Ultraviolet radiation (UVR) exposure is the most important environmental risk factor for NMSC. However, the precise relationship between UVR and the risk of NMSC is complex, and the relationship may differ by skin cancer type.MethodsA case–control study was conducted among Florida residents to investigate measures of patterns (intermittent vs. continuous) and timing (childhood vs. adulthood) of sunlight exposure in BCC and SCC. Participants included 218 BCC and 169 SCC cases recruited from a university dermatology clinic and 316 controls with no history of skin or other cancers.ResultsA history of blistering sunburn (a measure of intermittent sunlight exposure) was associated with both BCC (OR = 1.96, 95% CI = 1.27-3.03) and SCC (OR = 2.02, 95% CI = 1.22-3.33). Additionally, having a job in the sun for ≥3 months for 10 years or longer (a measure of continuous sunlight exposure) was also associated with both BCC and SCC in our study population. With the exception of younger age at first blistering sunburn, measures of younger age at sunlight exposure tended to be associated with SCC, but not BCC risk.ConclusionsResults from the current study suggest that sunlight exposure is associated with both BCC and SCC risk regardless of the pattern in which the exposure was received (i.e. intermittent vs. continuous). The data also suggest that sunlight exposure at a younger age may be more important for SCC but not BCC, however additional studies are needed to further characterize sunlight exposure-response relationships in different types of NMSC.

Rapid frozen section immunostaining of melanocytes by microphthalmia-associated transcription factor.

Mohs micrographic surgery (MMS) has increasingly become an accepted therapy for melanoma in situ on chronically sun damaged skin (CSDS). However, melanocytes are difficult to locate in frozen material on hematoxylin and eosin. In addition, determining the cut-off between the melanoma and the “atypical melanocytic hyperplasia” in CSDS can be challenging in frozen or formalin-fixed paraffin-embedded sections, with or without immunohistochemistry (IHC). In this article, we report the use of a rapid, 35-minute protocol using microphthalmia-associated transcription factor (MITF) IHC for identifying melanocytes in frozen tissue for its potential use in MMS. In contrast to melanoma antigen recognized by T cells (MART-1), MITF is a nuclear stain, which simplifies identification of melanocytes and quantification of melanocytic parameters. In this study, MITF IHC in frozen sections yielded equivalent melanocyte nuclear diameter and density measurements compared with formalin-fixed paraffin-embedded sections. Nuclear diameter measurements obtained with MITF were similar to that previously reported with MART-1, but the melanocyte density figures were lower. Reliable labeling of melanocytes in frozen sections required the use of diaminobenzidine (DAB) chromogen with Giemsa counterstaining and a buffer devoid of surfactant. Our experience with MITF IHC indicates that it is a dependable immunostain in frozen sections, and may prove to be useful in MMS as an adjunct to hematoxylin and eosin and MART-1 IHC for interpretation of margins for melanoma in situ on CSDS.

Multiple keratoacanthomas arising in the setting of sorafenib therapy: novel chemoprophylaxis with bexarotene.

Address correspondence to Basil S. Cherpelis, MD, Department of Dermatology, University of South Florida, 12901 Bruce B. Downs Boulevard, MDC 79, Tampa, FL 33612. E-mail: bcherpel@hsc.usf.edu as two cycles of paclitaxel and carboplatin chemotherapy and four cycles of docetaxel and carboplatin. Subsequently, her serum CA-125 level returned to normal, indicating remission of the cancer. However, 5 months after completing chemotherapy, her serum CA-125 level again elevated to 95 from a nadir of 5, indicating that cancer had returned. A PET/CT scan confirmed multiple hypermetabolic peritoneal masses as well as right inguinal nodal activity. An excisional biopsy was performed on the right inguinal node, and histology disclosed metastatic adenocarcinoma. The patient was referred to our institute for consideration of a clinical trial. In late 2007, she began a clinical trial using a combination of carboplatin, paclitaxel, and sorafenib. The patient continued her monthly follow-up meetings in the Gynecologic Oncology Program. Three months after beginning treatment with carboplatin, tamoxifen, and sorafenib, she reported a rapidly enlarging, non-healing erythematous papule on her left nares and presented to dermatology for evaluation. By this time, she had developed a similar lesion on the left leg (Fig 1). Histologic examination of these lesions revealed well-demarcated crateriform squamous proliferations with a central keratin plug consistent with kerIntroduction Sorafenib is a novel antineoplastic drug that targets multiple intracellular and cell-surface kinases, inhibiting tumor cell proliferation and angiogenesis. While it is currently approved to treat metastatic or unresectable renal cell or hepatocellular carcinoma, studies are underway to evaluate its utility in other solid organ tumors. We present a case of a 61-year-old woman with metastatic ovarian cancer and no history of skin cancer who developed multiple keratoacanthomas while on a clinical trial using sorafenib. Surgical excision was performed palliatively for large and symptomatic lesions. Because of the patient’s terminal condition and unwillingness to undergo multiple surgical procedures, we instituted chemoprophylaxis with oral bexarotene. After two months of therapy, the patient has reported partial regression of some of the tumors and has not developed any new lesions. This is the second known report of multiple keratoacanthomas arising in the setting of sorafenib therapy.1 In addition, this is the first known report of using bexarotene for chemoprophylaxis or treatment of keratoacanthomas.

Immunohistochemical stains in Mohs surgery: a review.

BACKGROUND During Mohs surgery, there are instances in which residual tumor cells may be difficult to detect, thereby increasing the risk of incomplete excision and tumor recurrence. It is possible to employ immunohistochemical techniques as an adjunct to routine hematoxylin and eosin staining to aid in ensuring negative margins. OBJECTIVE To review the literature regarding the use of immunostains in Mohs surgery. RESULTS Various immunostains have proved useful in detecting tumor cells in various malignancies, including melanoma, basal cell carcinoma, squamous cell carcinoma, dermatofibrosarcoma protuberans, extramammary Pagets disease, primary cutaneous mucinous carcinoma, granular cell tumor, and trichilemmal carcinoma. CONCLUSIONS In this article, we review immunohistochemical stains that have been employed in Mohs micrographic surgery and evaluate their utility in enhancing detection of residual tumors with respect to tumor type, particularly in situations in which detection of residual tumor may be difficult. The authors have indicated no significant interest with commercial supporters.