Prognostic value of TERT promoter mutations in conjunctival melanomas in addition to clinicopathological features.

AIMS: To evaluate the prognostic value of clinical, histopathological and molecular features and to relate different treatment modalities to clinical outcome in conjunctival melanomas (CM). METHODS: Retrospective review of clinical, histopathological and BRAF V600E and telomerase reverse transcriptase (TERT) promoter mutation status and treatment modalities, correlated to recurrence and metastasis in 79 patients with CM, diagnosed between 1987 and 2015 in three tertiary referral centres in the Netherlands and Belgium. RESULTS: Out of 78 evaluable patients, recurrences occurred in 16 patients and metastasis in 12 patients (median follow-up time 35 months (0-260 months)). Tumour thickness >2 mm, pT status, the presence of epithelioid cells, ulceration and mitoses was significantly correlated with metastasis (p value 0.046, 0.01, 0.02, 0.001 and 0.003, respectively). Furthermore, CM frequently harbour BRAF V600E and TERT promoter mutations (29% and 43%, respectively). TERT promoter mutations were correlated to shorter metastasis-free survival (p value 0.002). No significant correlation was found for clinical parameters and metastatic disease. Palpebral, forniceal and caruncular melanomas were more prone to develop recurrences (p value: 0.03). Most CM were treated with excision with adjuvant therapy. CONCLUSION: In line with the recommendations in the Eighth Edition of the American Joint Committee on Cancer Staging for CM, the pathology report should include information about pT status, tumour thickness, presence of epithelioid cells, ulceration and mitoses. Furthermore, information about the presence of a TERT promoter mutation and BRAF V600E mutation is of interest for therapeutic decision making. The presence of a TERT promoter mutation is correlated to metastatic disease.

Floating pigmented vitreous cyst.

Case report of a floating pigmented vitreous cyst.

Epistaxis or epiphora as a sign for extension of a conjunctival melanoma. A series of six patients with nasolacrimal recurrence.

PURPOSE: To characterise malignant conjunctival melanomas with extension and recurrence in the nasolacrimal system. METHODS: Localisation of the primary tumour and recurrences of 210 conjunctival melanomas treated in The Netherlands were reviewed for orbital and nasal tumours (1978-2008). Based of these cases and literature data, characteristics for nasolacrimal system extension and metastasis were reviewed. RESULTS: Six patients (3%) showed a recurrence of the primary conjunctival melanoma in the nasolacrimal system. Two of the six primary tumours were limbal tumours; the other four were diffuse tumours involving the fornix. In all six patients, the primary conjunctival melanomas were associated with primary acquired melanosis. During the follow-up period (11.6±3 years, range 3.4-28.5 years, median 8.7 years) two patients developed metastases and died. CONCLUSIONS: Patients should be advised to contact their treating ophthalmologist in the case of symptoms of epiphora, nose obstructions and epistaxis, especially non-bulbar and diffuse cases associated with primary acquired melanosis.

Cytotoxic effect of sodium hypochlorite 0.5% (NaOCl) on ocular melanoma cells in vitro.

PURPOSE: Excision with or without adjuvant cryotherapy or brachytherapy is the treatment of choice in conjunctival melanoma. Adjuvant rinsing with alcohol or sodium hypochlorite peroperatively (Dakin's solution) is used in some centers to prevent seeding of melanoma cells. The purpose of this research is to compare the cytotoxicity of sodium hypochlorite (NaOCl) with other potential cytotoxic solutions in the treatment of conjunctival melanoma. METHODS: Three uveal melanoma cell lines (OCM8, Mel285, and Mel270) and one conjunctival melanoma cell line (CM2005.1) were tested in a proliferation test (CellTiter 96 AQueous One Solution Cell Proliferation Assay, Promega, Madison, WI). The 96-well plates were coated with melanoma cells and treated with sodium hypochlorite 0.5%, sodium bicarbonate (1.4% and 8.4%), ethanol 99%, or sodium chlorite during 3, 5, or 15 minutes. Each solution was tested in several dilutions. RESULTS: In all cell lines, no surviving cells were observed after treatment of 3 minutes with sodium hypochlorite. Ethanol 99% had a similar effect. A reduction of 70% of viable cells could be reached using sodium bicarbonate 1.4% or 8.4%. Water reduced the amount of viable cells by 40%. CONCLUSIONS: Sodium hypochlorite is cytotoxic for melanocytic cells in vitro. Its use may reduce local seeding of tumor cells and may decrease metastasis after extirpation of an extended ocular tumour. Further in vivo evaluation of sodium hypochlorite is required.

Impression cytology of melanocytic conjunctival tumours using the Biopore membrane.

PURPOSE: To compare a new Biopore membrane impression cytology method with the routinely used exfoliative cytology in patients with a melanocytic lesion of the conjunctiva. METHODS: Sixty-eight consecutive patients with a conjunctival melanocytic lesion underwent Biopore membrane impression cytology as well as exfoliative cytology. A histologic sample was also available in 26 cases. All Biopore samples were stained immediately with RAL 555. Both Biopore and exfoliative cytology samples were assessed by two cytopathologists and graded into four different categories of atypia. RESULTS: Twenty-three out of 26 Biopores and 20 out of 24 for the exfoliative smears correlated with the corresponding histologic sample. Biopore cytology resulted in higher numbers of cells with a greater density compared to exfoliative cytology. CONCLUSIONS: Biopore cytology can be used for cytologic sampling of conjunctival melanocytic lesions. Because of the larger amount and higher density of cells obtained with the Biopore membrane, interpretation by a pathologist is easier and faster. Sampling of the fornix, caruncula, and ocular material in children is difficult with the Biopore method, and exfoliative cytology seems to be the favorable test in those situations.

S-100B protein and melanoma inhibitory activity protein in uveal melanoma screening. A comparison with liver function tests.

AIM: Our purpose was to determine whether S-100B or melanoma inhibitory activity (MIA) concentrations in the serum of patients with large uveal melanomas were better markers for the presentation of metastases than liver function tests. We also investigated whether increased marker levels were related to known clinical and histopathological prognostic parameters. METHODS: Total S-100B (A1B + BB) and MIA concentrations were measured in the sera from 104 patients with uveal melanoma prior to enucleation and in the sera from 50 healthy controls. Concentrations were also determined in the sera from 30 patients with known uveal melanoma metastases. Liaison Sangtec 100, an automated immunoluminometric assay measuring the total S-100B, and Roche MIA ELISA were used to quantify these proteins in serum. Results were compared with liver function tests [alkaline phosphatase, lactate dehydrogenase (LD), aspartate aminotransferase, alanine aminotransferase and gamma-glutamyl transpeptidase]. RESULTS: The mean S-100B and MIA concentrations were significantly higher in patients with metastases compared to melanoma patients without metastases. At the time of enucleation, S-100B and MIA were not prognostic for metastases in uveal melanoma, but S-100B and LD were the best tests to predict the occurrence of metastatic disease during the follow-up period. CONCLUSIONS: In our study, the S-100B and MIA serum concentrations were not correlated with any tested established prognostic parameter. S-100B and LD showed better performance in identifying melanoma metastases than gamma-glutamyl transpeptidase and MIA. A prospective follow-up study is needed to evaluate S-100B and MIA in identifying early micrometastasis in uveal melanoma.

Screening for conjunctival melanoma metastasis: literature review.

Local tumour control in conjunctival melanoma has improved in recent years. However the tumour-related death rate of these patients is still 14% at 5 years up to 33% at 15 years. With the introduction of sentinel node biopsies for conjunctival melanomas with a poor prognosis and screening for locoregional and distant metasases prognosis might be improved.

Immunophenotypic markers to differentiate between benign and malignant melanocytic lesions.

BACKGROUND/AIMS: The authors investigated the expression of S100A1, S100A6, S100B, MelanA, and CEA in conjunctival naevi, primary acquired melanosis (PAM), conjunctival melanoma, and uveal melanoma in order to assess their potential usefulness in the pathological differential diagnosis of these entities. METHODS: Paraffin embedded sections of 18 conjunctival naevi, 14 PAM, 16 conjunctival melanomas, and 20 uveal melanomas were immunostained for S100A1, S100A6, S100B, MelanA, and CEA, and expression was scored semiquantitatively. RESULTS: Expression of S100A1 differed significantly between conjunctival naevi and conjunctival melanoma, with percentages of positive cells of 30.6% and 71.4%, respectively. Conjunctival melanomas had high average scores for S100A1 and S100B (71.4%, 62.9%, respectively), while uveal melanomas also had high S100A1 but low S100B scores (88.5%, 18.5%, respectively). MelanA was highly variable; naevi and uveal melanoma had higher average scores than conjunctival melanoma. CEA was hardly detectable in all four groups. CONCLUSION: S100A1 seems to be a possible candidate to differentiate conjunctival naevi from conjunctival melanoma. S100B seems to differentiate between uveal melanoma and conjunctival melanoma. However, the study size was small and therefore the data have to be confirmed by others.

Screening for uveal melanoma metastasis. Literature review.

Local tumour control in uveal melanoma has improved in the last decades. However, 5-year mortality due to metastases from large uveal melanomas remains high. Recently both isolated liver perfusion therapy and chemotherapy have reached encouraging results in improving metastasis survival. As such screening at an early stage, especially for liver metastases, becomes imperative.