Personalized and targeted mutational analysis of multiple second primary melanomas under kinase inhibitors.

BACKGROUND: Second primary melanomas (SPMs) are new developed primary melanomas occurring in a subset of patients affected by BRAF-mutated metastatic melanoma during treatment with BRAF-inhibitors. A drug-induced paradoxical activation of mitogen-activated protein kinase (MAPK) signaling pathway in BRAF-wild type/RAS-mutated cells have been proposed as a possible molecular mechanism but data on the mutational status of SPMs are lacking. In order to better understand genetic alterations affecting the biological mechanism of SPMs, we performed a personalized and targeted next-generation sequencing analysis of a patient affected by metastatic melanoma who developed multiple SPMs during treatment with encorafenib (LGX818). METHODS: Using a cancer panel of 50 genes for solid tumors enriched with a custom panel of 10 genes specifically involved in melanoma pathogenesis, we analyzed the primary melanoma, two SPMs, one benign compound nevus and the normal DNA extracted from blood lymphocytes of the patient. RESULTS: We identified HRAS Q61 somatic mutation in one SPM developed in a pre-existing nevus. In the primary melanoma, besides the BRAF mutation, we identified the clinically actionable IDH1 R132C somatic mutation. Both SPMs were BRAF wild type. The patient harbors the recently recognized pathogenetic germline variant KDR Q472. We observed that mutations detected in tumor samples involving genes related to melanoma pathogenesis (TP53, PIK3CA, FGFR3, ATF1, KIT, HRAS and MAP2K2) were present in heterozygosis in the germline status of the patient. CONCLUSIONS: Our results support the paradoxical mechanism of MAPK pathway for SPMs under BRAF inhibitors. Moreover, they suggest that targeted mutational assessment based on matching somatic and germline analysis represent a promising approach to detect the neoplastic landscape of the tumor and to identify most accurate treatment in metastatic melanoma patient.

Expression of estrogen receptors in Spitz and Reed nevi.

BACKGROUND: Estrogens play a key role in the skin. They are associated with an increased production of melanin, proliferation of melanocytes, increased skin thickness and increased cutaneous vascularization. Spitz and Reed nevi are acquired melanocytic lesions that generally develop during childhood or adolescence, a period of changes in sex hormones background. Our study project aimed at investigating, through immunohistochemical analysis, the expression levels of ERß receptors and their expression patterns (cytoplasmic or nuclear) in Spitz and Reed nevi. METHODS: In our study, we collected a total of 86 melanocytic lesions of patients: of these, 16 were common nevi, 23 were Spitz nevi, 18 were Reed nevi and 29 were melanomas. Expression curves for estrogen receptors were constructed using the Kaplan-Meier method and compared using a log-rank test. Statistical analysis was performed using MedCalc® (MedCalc Software, Ostend, Belgium). Immunohistochemical analysis on all histological sections of nevi and melanomas was performed to evaluate the expression levels of of ERß and their expression patterns (cytoplasmic or nuclear). The agreement between the operators was calculated using Fleiss κ values. RESULTS: The correlation between immunoreactivity for the ß-estrogen receptor and the sex of patients with Spitz and Reed nevi showed that immunoreactivity was higher in male patients. The correlation between ß-estrogen receptor immunoreactivity and patient age for Spitz and Reed nevi showed no statistically significant correlation. Correlation between immunoreactivity for the ß-estrogen receptor and histotype: Spitz and Reed nevi showed a high intensity, while in common nevi and in melanomas the immunoreactive was low. The correlation between receptor immunoreactivity for ß estrogens and Breslow thickness in melanomas indicated that Breslow thickness of non-immunoreactive melanomas for ERß was much higher than those showing high immunoreactivity for this receptor. CONCLUSIONS: Spitz and Reed nevi express a higher immunoreactivity for estrogens than common nevi and melanomas, especially those with a high Breslow thickness; and immunoreactivity is higher in younger age groups.

Basal cell carcinoma thickness evaluated by high-frequency ultrasounds and correlation with dermoscopic features.

BACKGROUND: Basal cell carcinoma (BCC) is the most common skin cancer, and it can be easily treated by surgery or by various other physical modalities and topical chemotherapy. For metastatic, locally advanced BCC and for cancers that cannot be removed by surgery, systemic drugs known as hedgehog pathway blocker are used. High-frequency ultrasound (HFUS) is a non- invasive technique used in diagnosis of some skin cancers. It has proven potentially useful for BCC management. In this study we used high frequency ultrasounds to evaluate BCCs' thickness and the correlation with dermoscopic features. METHODS: We examined 86 basal cell carcinomas with dermoscopy and with high-frequency ultrasound. The main patterns identified by ultrasound were linear, ellipsoid and non-specific or undefined. Patients were divided by sex and age. The BCCs were grouped by anatomic location. Finally, we recorded specific dermoscopic features of BCCs noting their presence/absence in lesions overall and in each of four quadrants. Then the lesions were excised, and histological examination was made with definition of tumor thickness (in mm). RESULTS: In our study, two main echographic patterns were described: linear, associated with superficial BCC, and ellipsoid, found primarily in nodular variants. However, a small percentage of lesions have otherwise non-specific patterns. We observed a significant correlation between echographic tumor thickness and histotype. We observed high concordance between histological tumor thickness and ultrasounds. Also, dermoscopic criteria as large branching and blue ovoid nests were significantly associated with heightened histologic and echographic assessments of tumor thickness. CONCLUSIONS: Our study confirmed the utility of ultrasound in the diagnosis of BCCs and for the first time we have correlated ultrasounds' patterns with dermoscopy and tumor thickness.

Upper eyelid Merkel cell carcinoma treated with neoadjuvant chemotherapy and surgical excision.

Merkel cell carcinoma is a rare cutaneous carcinoma, featured by an aggressive clinical course and a mortality rate of 28% at 2 years. A 71-year-old female was affected by a 4.1-cm-wide locally advanced Merkel cell carcinoma of the upper eyelid, previously misdiagnosed as chalazion, with involvement of the extraocular muscles. Although the tumor showed a macroscopic spontaneous regression in size after the incisional biopsy, the mass was treated with neoadjuvant chemotherapy and surgical excision. Good functional and aesthetic result with preservation of the eyeball and absence of tumor recurrence were achieved at 3-year follow-up. In our experience, the combination of the inflammatory cascade due to the incisional biopsy and neoadjuvant chemotherapy led to the regression of a locally advanced large Merkel cell carcinoma of the eyelid.

Beyond appearance: An unusual manifestation of isolated oral secondary syphilis.

Syphilis is a sexually acquired chronic infection caused by Treponema pallidum and is characterized by a variety of clinical manifestations. The secondary stage of the disease results from the hematogenous and lymphatic dissemination of treponemes after a few weeks or months, and it is characterized by recurrent activity of the disease, with muco-cutaneous as well as systemic manifestations. Mucosal lesions range from small, superficial ulcers that resemble painless aphthae to large gray plaques, and they are generally associated with systemic manifestations of the disease. The exclusive asymptomatic oral localization not associated with general manifestations is uncommon but may actually be unrecognized and under-reported. We report a case of isolated oral manifestation as the unique presentation of secondary syphilis.

Efficacy of Ingenol Mebutate in the Treatment of Actinic Keratoses: A Pre- and Posttreatment Dermoscopic Comparative Analysis.

Actinic keratosis (AK) is a common skin lesion in adults which usually occurs on chronically photoexposed areas and considered as a precancerous lesion or a superficial squamous-cell carcinoma. Many classifications have been proposed and its diagnosis is generally clinical but, sometimes, its wide variety of presentations can make diagnosis difficult, even among expert observers. The malignant potential of AKs imposes an early diagnosis and treatment in order to reduce morbidity and mortality, and, for the characterization of photodamaged skin, noninvasive diagnostic techniques, such as dermoscopy, have proved to be useful, while multiple therapeutic strategies, lesion-directed versus field-directed therapies, are available for the treatment of AKs. In this study, we evaluated the efficacy of ingenol mebutate for the treatment of AKs, with a particular focus on patients' compliance, correlating it to clinical and dermoscopic grading, pre- and posttreatment, of these lesions. Fifty-two enrolled patients with AKs received treatment with ingenol mebutate gel (0.015% for face and scalp; 0.05% for trunk and extremities) and multiple dermatological evaluations. End points of the study were complete and partial clearance of clinically visible AKs on day 90. All acquired data were recorded and statistical analyses were performed. Univariate and multivariate analyses were used to identify possible predictive factors. We retrospectively analyzed patient-related and lesion-related factors to identify which variables, among age, gender, lesion site, pain, LSR score, and pretreatment clinical and dermoscopic grading, could independently predict the response to ingenol mebutate treatment. Our findings showed that pretreatment dermoscopic grade II represents an independent predictive factor of the efficacy of ingenol mebutate therapy (OR=14.78, 95% CI: 1.83-119.59, P=0.012) and that response rates differ on the basis of the treated anatomical sites (OR=0.16, 95% CI: 0.03-0.85, P=0.031). Data from this study provide evidence that ingenol mebutate gel is an effective treatment for AK, with relative ease of use, short exposure, and rapid resolution of local reactions, benefits contributing to high adherence of this therapy. Moreover, dermoscopic analysis of skin lesions offers more information than clinical evaluation alone and can be helpful in identifying different groups of AKs, thus selecting the adequate therapeutic choice.

Cutaneous candidiasis caused by Candida albicans in a young non-immunosuppressed patient: an unusual presentation.

Candidiasis is a fungal infection caused by yeasts that belong to the genus Candida. There are over 20 species of Candida yeasts that can cause infection in humans, the most common of which is Candida albicans. Candida yeasts normally reside in the intestinal tract and can be found on mucous membranes and skin without causing infection. However, under immunocompromised conditions, Candida can cause significant infections in susceptible patients. Herein, we report a peculiar presentation of a C. albicans cutaneous infection in an immunocompetent young subject. This case widens our knowledge on the C. albicans infections both in terms of host susceptibility and cutaneous manifestations.

Insulin-like-growth-factor-binding-protein-3 (IGFBP-3) contrasts melanoma progression in vitro and in vivo.

Insulin-like-factor-binding-protein 3 (IGFBP-3) is known to modulate the activity of insulin-like growth factors (IGFs) besides having a number of IGF-independent effects on cell growth and survival. IGFBP-3 has been reported to decrease significantly in the blood serum of patients affected by certain cancers. In the present work, we have evaluated the levels of IGFBP-3 in the blood serum and tissues of patients affected by cutaneous melanoma, showing that loss of IGFBP-3 from both is strongly correlated with disease progression and reduced survival. In vitro treatment with IGFBP-3 of human and murine metastatic melanoma cell lines specifically inhibited the cells' migratory and invasive behaviour, inducing up-regulation of melanocytic differentiation markers such as tyrosinase activity and melanin content. A molecular analysis of the cellular pathways transducing the effect of IGFBP-3 implicated the Akt-GSK3ß axis. Moreover, administration of IGFBP-3 in vivo to SCID mice inoculated with human metastatic melanoma cells strongly reduced or completely inhibited tumor growth. In summary, IGFBP-3 appears to exert a specific inhibitory effect on melanoma growth and dissemination, suggesting that it may qualify as a useful therapeutic agent in melanomas and perhaps other cancers, at the least as a valid adjuvant therapy during treatment with conventional anti-tumoral drugs.

Melanoma with unknown primary: report and analysis of 24 patients.

In the literature, there are some papers reporting on patients with metastatic melanoma from an unknown primary lesion (MUP). The pathogenesis of this phenomenon and the prognosis of these patients are still debatable. Therefore, we reviewed our casistics on MUP patients. We identified 24 MUP patients out of all patients registered into a melanoma database from June 1996 to June 2011. The incidence was 1.4%. We compared the survival rate of all patients with MUP stage III-IV with all patients with metastatic melanoma known primary (MMKP) stage III-IV observing a clear survival improvement for MUP patients in front of MMKP patients (p<0.01). In a second instance, we compared stage III MUP patients with only lymph nodal involvement with stage III MMKP patients with only lymph nodal involvement, and again we found statistically significant better survival for MUP patients (p<0.05). In this retrospective study, the number of lymph nodes involved (p=0.8), the sex (p=0.9), and S100 value (p=0.2) were not statistically relevant for prognosis. The better prognosis for these patients is very similar to better survival rate for metastatic melanoma patients and vitiligo. This correlation may be in accord with the hypothesis of a regression of primary lesion by immunological system of the host and also the median age of patients at the time of diagnosis, commonly older than melanoma patients, may correspond to a long period of immunological interferences between the host and the melanoma disease.

Correlation between insulin-like growth factor binding protein-3 serum level and melanoma progression.

BACKGROUND: Insulin-like growth factor (IGF) binding protein (IGFBP)-3 is the main carrier of circulating IGFs and the main modulator of their activity. IGFBP-3 controls cellular availability of IGFs, which cannot exert their pro-proliferative activity while bound to IGFBP-3. Proteolysis of IGFBP-3 is one mechanism to control IGF release. A reduction of serum IGFBP-3 levels and the associated increased availability of IGFs may represent a strategy whereby melanoma increases its metastatic potential. OBJECTIVE: The aim of our study was to evaluate the correlation between the IGFBP-3 serum level and melanoma stage. METHODS: The study included 41 patients, 24 male and 17 female, with median age of 60 years (range 24-80), affected by cutaneous melanoma. Blood samples were taken from each patient and IGFBP-3 serum levels were measured using Western blot analysis with commercial antibodies. Values were normalized using commercial IGFBP-3. RESULTS: The statistical analysis showed that full-size, glycosylated IGFBP-3 concentrations were significantly lower in the sera of patients with stage IV melanoma. Low serum levels of IGFBP-3 correlated with both disease progression and presence of disease at the time of sample collection. In patients who underwent follow-up visits with further collections of blood samples, the concentrations of glycosylated IGFBP-3 decreased only in those who showed progression of disease. LIMITATIONS: Our study shows only preliminary results on a limited number of patients. CONCLUSION: We demonstrate that there is a significant inverse correlation between the serum concentration of full-size, glycosylated IGFBP-3 and disease progression in patients with melanoma.

Predictive factors for false negative sentinel lymph node in melanoma patients.

BACKGROUND: Sentinel lymph node biopsy (SLNB) represents a useful tool for staging melanoma patients. However false-negative SLNB are reported in the literature. OBJECTIVE: The aim of our study is to identify predictive factors for false-negative SLNB in melanoma patients. MATERIALS AND METHODS: We conducted a retrospective analysis on 316 melanoma patients who underwent SLNB and were followed up at the Department of Dermatology and Plastic Surgery of University of Rome "Sapienza" from March 1994 to June 2008. RESULTS: In our patients, SLNB was positive in 35 cases (11.07%) whereas it was negative in 281 cases (88.93%); 12/316 patients (3.8%) had positive SLNB and positive therapeutic lymph node dissection (TLND); 23/316 (7.28%) patients had positive SLNB and negative TLND; 266/316 (84.18%) patients had negative SLNB but without subsequent metastases in the SLN site; 15/316 (4.74%) patients had negative SLNB, but with subsequent metastases in the same SLN site (false-negative patients). Among the different prognostic factors, only ulceration was the main predictive factor for false-negative SLNB, according to statistical analysis (p=.0420). CONCLUSION: Our data confirm that SLNB is a useful technique for staging melanoma patients. However, in patients with negative SLNB, a closer follow-up is recommended when ulceration is present. The authors have indicated no significant interest with commercial supporters.

Amoxicillin-clavulanic acid-induced linear immunoglobulin A bullous dermatosis: case report and review of the literature.

BACKGROUND: Linear immunoglobulin A bullous dermatosis (LABD) is an autoimmune subepidermal blistering disease, rarely induced by drugs. METHODS: We describe a case of a 47-year-old man who developed a severe blistering eruption after therapy with amoxicillin-clavulanic acid. RESULTS: Histopathologic examination and direct immunofluorescence were consistent with a diagnosis of LABD. Therapy with dapsone controlled the disease but, after sun exposure, there was a worsening of the illness. CONCLUSION: To our knowledge, this is the first case of amoxicillin-clavulanic acid-induced LABD in an adult.

The reticular point of view in dermatoscopy.

BACKGROUND: Different dermatoscopic algorithms have been developed to evaluate pigmented lesions of the skin, with pattern analysis being the most commonly used. We sought to develop and evaluate a diagnostic scoring system, the reticular point of view, to distinguish common melanocytic nevus from dysplastic nevus and from melanoma. METHODS: We analyzed 1543 pigmented lesions, most of which contained a pigment network, and assessed the presence of linear extensions, thickened or hyperpigmented lines, areas of abrupt cut off, and areas with large "holes." We then conducted statistical analyses on these parameters to verify which of these exerted the most influence on the dermatologist's decision to surgically excise the lesion and to verify which feature was most linked to histopathologic signs of dysplasia or malignancy. RESULTS: Among the lesions excised, histopathologic examination revealed 33 (10.28%) melanomas. Among these, 25 (75.75%) showed an asymmetric distribution of the pigment network, whereas 20 (60.6%) showed a linear extension of it. The analysis of our data showed that all the evaluated criteria were statistically significant and played an important role in the dermatologist's decision to perform surgical excision of the lesion. Regarding the diagnosis of melanoma, only the presence of large holes, areas of abrupt cut off, and linear extensions revealed statistical significance. LIMITATIONS: Reticular point of view may be useful only in lesions with a large pigment network. It is not applicable in nodular, thick, and amelanotic melanomas that are usually lacking in pigment network. CONCLUSION: Although pattern analysis represents the most effective analytical method in dermatoscopy, our scoring system may be useful to distinguish between benign lesions, which need only clinical follow-up, and malignant lesions, which need surgical excision.

Management of skin ulcers in a patient with mycosis fungoides.

We present a patient with a cutaneous T-cell lymphoma/mycosis fungoides (CTCL/MF) followed for more than 10 years. After several different aggressive treatments to control progression of CTCL/MF, the patient developed several ulcerated tumors on the abdomen and limbs. Specific systemic antibiotic therapy failed to treat skin infection. While treating the stage III CTCL with polychemotherapy, we used an active colloidal hydrogel topically to manage wound healing and to treat and prevent potential sources of sepsis. After 11 weeks of treatment we observed complete cicatrization of ulcerated tumors. We reported on this case to describe the importance of a correct management of skin ulcers in immunosuppressed patients in order to avoid possible systemic spread of infection which represents the major cause of death in these patients.

Complete remission of brain metastases in three patients with stage IV melanoma treated with BOLD and G-CSF.

BACKGROUND: Brain metastases are the most life-threatening among the secondary localizations of melanoma for their unresponsiveness to the surgical, radiotherapeutic and/or chemotherapeutic treatments. METHODS: Accidentally, we observed a complete response (CR) in a patient undergoing chemotherapy with bleomycin, vincristine or Oncovin, CCNU or lomustine, dacarbazine (BOLD) regimen for metastatic melanoma including brain metastases, who was also treated with G-CSF to manage a concomitant leukopenia. After this observation, seven more patients with stage IV melanoma with brain metastases were treated with BOLD regimen repeated every 6 weeks with administration of G-CSF in the intervals. RESULTS: Three patients presented CR (37.5%). Two patients stopped the treatment after two courses for evident progressive disease (25%). The other three patients showed stable disease (SD: 37.5%). Median duration of SD was 24 weeks. Among the eight patients, six (75%) achieved clinical benefit. Median time to progression was 8.5 months (range 0-74+ months). Median survival was 12.5 months (range 4-74+ months). Two patients are still alive and disease-free after 74 and 57 months, respectively. CONCLUSION: We believe that the brilliant CR, the long duration of the disease-free intervals and the long survival in at least three of eight patients should encourage further research on BOLD with G-CSF for the treatment of advanced melanoma.