Primary Conjunctival Melanoma Research Articles

Lactate secreted by glycolytic conjunctival melanoma cells attracts and polarizes macrophages to drive angiogenesis in zebrafish xenografts.

Conjunctival melanoma (CoM) is a rare but potentially lethal cancer of the eye, with limited therapeutic option for metastases. A better understanding how primary CoM disseminate to form metastases is urgently needed in order to develop novel therapies. Previous studies indicated that primary CoM tumors express Vascular Endothelial Growth Factor (VEGF) and may recruit pro-tumorigenic M2-like macrophages. However, due to a lack of proper models, the expected role of angiogenesis in the metastatic dissemination of CoM is still unknown. We show that cells derived from two CoM cell lines induce a strong angiogenic response when xenografted in zebrafish larvae. CoM cells are highly glycolytic and secrete lactate, which recruits and polarizes human and zebrafish macrophages towards a M2-like phenotype. These macrophages elevate the levels of proangiogenic factors such as VEGF, TGF-β, and IL-10 in the tumor microenvironment to induce an angiogenic response towards the engrafted CoM cells in vivo. Chemical ablation of zebrafish macrophages or inhibition of glycolysis in CoM cells terminates this response, suggesting that attraction of lactate-dependent macrophages into engrafted CoM cells drives angiogenesis and serves as a possible dissemination mechanism for glycolytic CoM cells.

Post-operative interventional radiotherapy (brachytherapy) in advanced ocular surface and eyelid tumors as an alternative to surgical retreatment.

The main purpose of treatment of advanced ocular surface and periocular malignant tumors is to eradicate the tumor while trying to preserve visual function and aesthetics. Our purpose is to describe the outcome of a retrospective case series of 10 patients with advanced ocular surface and periocular tumors treated surgically in first instance and then with postoperative interventional radiotherapy (IRT/Brachiterapy). We describe the clinicopathological features, treatments and outcome, in a retrospective case series of 10 patients with advanced tumors involving ocular surface (staging ≥ T2) and eyelids (staging ≥ T3), with involvement of periocular and/or orbit tissues. Patients were first surgically treated, most of them with incomplete excision, and then underwent a post-operative interventional radiotherapy (IRT/Brachytherapy) as an alternative to more invasive and disfiguring surgical retreatment. Tumor location, risk factors, staging, histological features, and follow-up timing were analyzed. Three patients had advanced eyelid basal cell carcinomas, 2 patients were diagnosed with eyelid and conjunctival squamous cell carcinomas, 3 as sebaceous carcinomas, and 2 as primary conjunctival melanomas. The mean follow-up time from IRT to last clinical follow-up was 58.6 weeks, range 28.4-168 (median 43.65, IQR 28.9-72.9). Two patients - one with ocular surface SCC, the other with conjunctival melanoma - had a local recurrence 23.4 and 40,9 weeks after IRT, respectively. An overview of the current knowledge on adjuvant or post-operative IRT is also provided. IRT can be considered an effective therapeutic option to avoid more invasive surgical retreatment in advanced tumors involving eyelids and ocular surface.

Conjunctival melanoma: A 20-year survey in a comprehensive medical center

To better understand population-specific tumor characteristics and behavior of conjunctival melanoma in Asian. A retrospective cohort enrolled patients with primary conjunctival melanoma treated and followed up at Chang Gung Memorial Hospital (CGMH) in Taiwan between 1995 and 2015. Basic characteristics such as age, gender, tumor size, cell type, location, and TNM stage were recorded. Prognostic parameters included disease free interval, local recurrence, distant metastasis, and survival were analyzed. There were 20 patients enrolled in the study hospital between 1995 and 2015. All were histological proved by pathologists. Their mean age at diagnosis was 57.8±15.9 years. The mean follow-up time was 68.7±55.8 months. Mean tumor thickness was 6.5±5.9mm. Six patients developed local recurrence. Twelve patients had distant metastasis. Ten patients died from conjunctival melanoma. The 10 patients had greater mean tumor thickness (8.4±7.5mm) and shorter mean survival time (40.2±24.1 months). Tumor thickness was found as a prognostic factor for survival time (HR=1.15, P=0.01). Age, gender, T stage, and tumor location were not significantly associated with survival. Different tumor characteristics were found in this cohort. Higher metastasis and mortality rate could suggest a more aggressive disease pattern. Tumor thickness was indicated as a prognostic factor for survival time and was greater in size in cases with distant metastasis. Early and more invasive intervention with closely follow-up may be indicated in these cases.

Conjunctival melanoma is a molecular distinct mucosal melanoma with high expression of PD‐L1

AbstractPurposeConjunctival melanoma is a unique mucosal melanoma due to its location in the sunexposed conjunctiva. The purpose of this study was to contextualize conjunctival melanoma as a mucosal melanoma based on molecular profiling. This is the first study of its kind.Material and Methods12 primary conjunctival melanomas and 28 primary mucosal melanomas from non‐sunexposed sites (nasal cavity, vagina, and rectum) were subjected to molecular profiling using targeted next‐generation sequencing (Oncomine Comprehensive Cancer Panel) and NanoString gene expression profiling (PanCancer Pathways Panel, 770 cancer genes). Normal conjunctiva and mucosa were used as internal controls.ResultsGenetically, conjunctival melanomas harboured BRAF‐V600E mutations and NF1 mutations, whereas other mucosal melanomas harboured more unspecific mutations, such as atypical BRAF mutations and mutations in KIT. Non‐sunexposed mucosal melanomas frequently presented amplification of MYC. When comparing the gene expression profile of conjunctival melanoma and non‐sunexposed mucosal melanoma, 41 genes were found to be significantly deregulated. 24 genes were upregulated, whereas 17 were downregulated in conjunctival melanoma. Many of the deregulated genes were involved in immunological pathways when performing an in silico pathway analysis. Most intriguingly, programmed death‐ligand 1 (PD‐L1) presented a significant 6‐fold upregulation in conjunctival melanoma. This finding could be validated with immunohistochemistry (p = 0.041, Fisher’s exact test). A network analysis revealed that PD‐L1 was co‐expressed with 8 other genes.ConclusionsComprehensive molecular profiling on the DNA and mRNA level revealed that conjunctival melanoma is a distinct molecular mucosal melanoma characterized by a higher frequency of BRAF‐V600E mutations, low expression of MYC and a high expression of PD‐L1 and other genes involved in immune response. This finding is important, as this suggests a higher response to checkpoint inhibition in conjunctival melanoma metastases when compared to other mucosal melanomas.

Global microRNA profiling of metastatic conjunctival melanoma.

This study aimed to investigate the microRNA (miRNA) profile in primary tumors from conjunctival melanoma with and without subsequent metastatic spread along with their coupled distant metastases to identify miRNAs likely to be involved in metastatic progression. This observational study included 13 patients with metastatic conjunctival melanoma (follow-up: 1-39 years) treated at a Danish referral center. Twenty-five patients with nonmetastatic conjunctival melanoma (follow-up: 5-17 years) were included for comparison. Global miRNA profiling was performed with the Affymetrix GeneChip 4.1 microarray. Taqman qPCR arrays were used for validation. Significant differentially expressed miRNAs were defined as having a false discovery rate of less than 0.05. Primary conjunctival melanoma with and without subsequent metastatic spread clustered separately according to miRNA expression, and 15 miRNAs were found to have significant differential expression. Six miRNAs (hsa-miR-4528, hsa-miR-1270, hsa-miR-1290, hsa-mir-548f-4, hsa-mir-4278, and hsa-miR-34a-3p) were downregulated and nine miRNAs were upregulated (hsa-mir-575, hsa-miR-527, hsa-miR-518a-5p, hsa-miR-6759-5p, hsa-miR-8078, hsa-mir-4501, hsa-mir-622, hsa-mir-4698, and hsa-mir-4654) in primary conjunctival melanoma with subsequent metastatic spread. A comparison of primary conjunctival melanoma with their pair-matched metastases identified six significant differentially expressed miRNAs (hsa-miR-1246 and hsa-miR-302d-5p, hsa-mir-6084, hsa-miR-184, hsa-mir-658, and hsa-mir-4427). qPCR confirmed downregulation of hsa-miR-184 in the distant metastases when compared with the corresponding primary tumor. Primary conjunctival melanoma with and without subsequent metastatic spread separated clearly on the miRNA level when profiled with microarray-based methods. qPCR was able to replicate expression levels of one miRNA (hsa-miR-184) that was downregulated in metastases when compared with corresponding primary tumors.

Pigmentation of conjunctival melanoma recurrences and outcome

PurposeIn primary conjunctival melanoma (CoM), one of the characteristics that is associated with an increased risk of metastases and death is a lack of tumour pigmentation. The aim of this study was to investigate whether the degree of pigmentation of CoM recurrences relates similarly to clinical outcome.MethodsA data set of 177 patients with a CoM recurrence from the Wills Eye Hospital (USA) and the Leiden University Medical Center (The Netherlands) was analysed. The relation between clinical tumour pigmentation of the recurrences, the characteristics of the primary lesions and clinical outcome was investigated.ResultsIn 117 (66%) of 177 patients with a CoM recurrence, tumour pigmentation was known: 71 patients (61%) had recurrences with low pigmentation. Primary lesions had low pigmentation in 39% of cases, which is significantly different (p = 0.001). However, low tumour pigmentation of recurrences correlated with low tumour pigmentation of the primary lesion (p < 0.001). No association was observed between pigmentation of the recurrences and iris colour (p = 0.66). Low pigmentation of the recurrences was not significantly associated with an increased risk for metastases (HR 1.96, p = 0.12) or death (HR 1.79, p = 0.27), whereas primary tumours with low pigmentation did show a greater risk for metastases (HR 2.82, p = 0.016) and death (HR 2.90, p = 0.037).ConclusionsCoM recurrences are more often lightly pigmented compared to primary lesions. A correlation exists between the degree of pigmentation of primary and recurrent lesions, but recurrences can appear with any degree of pigmentation. Unlike primary CoM, the level of pigmentation of CoM recurrences is not related to metastasis or death.

Genetic Profiling of Primary Orbital Melanoma: An Analysis of 6 Cases with Clinicopathologic Correlation

To analyze the genetic profile of 6 cases of primary orbital melanoma with clinicopathologic correlation. Retrospective noninterventional study to analyze the genetic profile of 6 cases of primary orbital melanoma and to correlate the genetic findings with prognosis and clinicopathologic features. Inclusion criteria were patients with primary orbital melanoma with no evidence of primary eyelid skin, conjunctival, uveal, or remote melanoma at extraocular sites. The study involved 6 primary orbital melanomas from 6 patients. Four patients were exenterated and 2 had incisional biopsies performed. Clinical notes and radiologic records were assessed to ascertain clinical tumor behavior. Sections were stained with hematoxylin-eosin and exposed to immunohistochemistry for S100, MelA, HMB45, Sox10, and BAP1. Melanoma DNA was exposed to array comparative genomic hybridization to assess gross chromosomal copy number changes. Point mutation assessment and Sanger sequencing were performed for GNAQ, GNA11, BRAF, NRAS, pTERT, SF3B1, and EIF1AX. These were the presence of gross chromosomal copy number changes and the presence of mutations in GNAQ, GNA11, BRAF, NRAS, pTERT, SF3B1, and EIF1AX; the presence of metastases and time period between diagnosis and death from melanoma; and correlation between the tumor genetic profile and the clinical behavior of the tumor. One of the 6 cases was clinically associated with oculodermal melanocytosis. Of the 6 patients, 3 died of melanoma metastases and 1 of unrelated causes; 2 remain alive at last review. Three of the 6 cases were histologically associated with a benign precursor lesion. All melanomas expressed S100, MelA, HMB45, and Sox10. One patient showed loss of BAP1 nuclear staining. The most frequent chromosomal gains across the 6 cases, in order of frequency, were 6p, 8q, 17q, 6q, and 20p. The most frequently lost regions were 1p, 9p, 16q, and 17p. One patient showed monsomy 3 and gain of 8q (and showed the BAP1 loss). Mutations were found in GNAQ (1 case), GNA11 (1 case), SF3B1 (2 cases), NRAS (2 cases), and pTERT (2 cases). The data point to 2 genetic groups for primary orbital conjunctiva melanoma-like and a uveal melanoma-like group. A larger study would help confirm this suggestion.

Detection of mutations in SF3B1, EIF1AX and GNAQ in primary orbital melanoma by candidate gene analysis

BackgroundOcular melanoma is a rare but often deadly malignancy that arises in the uvea (commonest primary site), conjunctiva or the orbit. Primary orbital melanoma (POM) is exceedingly rare, with approximately 60 cases reported to date. Despite recent advances in our understanding of the genetics of primary uveal and conjunctival melanomas, this information is lacking for POM.MethodsDNA was extracted from 12 POM tissues, with matched germline DNA (where available). MLPA was conducted to detect chromosomal alterations and Sanger sequencing used to identify point mutations in candidate melanoma driver genes (BRAF, NRAS, KRAS, GNA11, GNAQ), and other genes implicated in melanoma prognosis (EIF1AX, SF3B1). Immunohistochemistry was performed to analyse BAP1 nuclear expression.ResultsMLPA detected copy number alterations in chromosomes 1p, 3, 6 and 8. Sequencing of melanoma driver genes revealed GNAQ (p.Q209L) mutations in two samples; although it is possible that these samples represent extraocular spread of an occult uveal melanoma. A recurrent mutation in SF3B1 (p.R625H) was observed in indolent, but not aggressive, tumours; a mutation in EIF1AX (p.N4S) was detected in one patient with non-aggressive disease.ConclusionsEIF1AX and SF3B1 mutations appear have a role in determining the clinical course of POM and detection of these changes could have clinical significance. Further in depth analysis of this rare group using differing ‘omic technologies will provide novel insights into tumour pathogenesis.

Expression of VEGF in human conjunctival melanoma analyzed with immunohistochemistry.

PurposeThe aim of this study was to examine the immunolocalization of VEGF-A and CD34, a marker of endothelial cells, in human conjunctival melanoma.MethodsThis study retrospectively analyzed primary conjunctival melanoma patients who underwent surgical resection of the tumor. All excised tissues were fixed with paraformaldehyde and embedded in paraffin, which were then submitted for immunohistochemistry with anti-VEGF and CD34 antibodies.ResultsThe study sample comprised 4 female and two male melanoma patients. The age of the patients ranged from 64 to 84 (average age, 73) years. Histopathology of the surgically resected tumor tissues demonstrated accumulation of polygonal atypical malignant cells producing melanin. Cytoplasmic immunoreactivity for VEGF was clearly observed in tumor cells of all six tumors. In contrast, CD34-positive endothelial cells were less marked in the melanoma tissues than in the adjacent noncancerous subconjunctival stroma.ConclusionVEGF immunoreactivity was observed in conjunctival melanoma tissues, in which endothelial cells were hardly observed. These results suggest that although VEGF is expressed, conjunctival melanoma is a hypovascular tumor.

Blue Nevi of the Ocular Surface: Clinical Characteristics, Pathologic Features, and Clinical Course

Blue nevus is a melanocytic tumor that is commonly found in the skin. Extracutaneous presentations, including the ocular surface, are rare. As such, the purpose of this study was to characterize the clinical features and clinical course of congenital melanocytic tumor (blue nevus) of the conjunctiva. Retrospective, noncomparative case series. Twenty-one patients with 23 blue nevi of the ocular surface that were excised surgically between 2000 and2016. Chart review of patients identified from a database search of the Florida Lions Ocular Pathology Laboratory records. Pathologic diagnoses were confirmed by 2 pathologists (S.R.D. and G.E.). All specimens were bleached and, tissue permitting, stained using SOX10 (MilliporeSigma, Darmstadt, Germany) and CD68 (Leica Biosystems, Nussloch, Germany). Clinical characteristics, pathologic features, and clinical course. Mean age of the population was 55±15 years; 71.4% (n= 15) were white and 57.1% (n= 12) were men. One patient had 3 lesions, for a total of 23 lesions examined. Clinically, 13 lesions were on the bulbar conjunctiva, 3 were on the tarsal conjunctiva, 3 were in the fornix, 2 were caruncular, 1 was episcleral, and 1 was at the limbus. Before excision, 8 patients were thought to have primary acquired melanosis, 4 with concern for primary conjunctival melanoma, and 1 thought to have metastatic disease from a plantar melanoma. Five lesions were thought to be benign, and in 8 patients, the lesions were identified incidentally after other ocular surgeries, with no diagnosis of the lesions before excision. Pathologic features were consistent with simple blue nevi in 21 lesions and cellular blue nevus in 2 lesions. No malignant transformations were noted in any patient over the mean 20.2-month follow-up period (range, 2 weeks-103 months). Blue nevus is a rare deeply pigmented congenital melanocytic lesion with a benign clinical course that can appear clinically similar to primary acquired melanosis or melanoma.

Treatment of conjunctival melanoma in a Dutch referral centre

AimsTo evaluate the treatment of conjunctival melanoma at a large Dutch referral centre and to make recommendations for clinical management.MethodsA retrospective review was performed of clinical and histological data of...

Satellite in transit metastases in rapidly fatal conjunctival melanoma: implications for angiotropism and extravascular migratory metastasis (description of a murine model for conjunctival melanoma)

Little information is currently available concerning loco-regional metastases such as satellite and in transit metastases and their natural history in conjunctival melanoma as compared to cutaneous melanoma. Angiotropism, a marker of extravascular migration of melanoma cells along vascular channels, often appears responsible for microscopic satellite, satellite and in transit metastases development in cutaneous melanoma. In addition, diffuse tissue microscopic satellites are correlated with widespread melanoma dissemination and death. Herein we report rapid conjunctival melanoma progression and a fatal outcome in four of five patients following recurrence as satellite in transit metastases. Five patients aged 31, 60, 63, 56, and 67 years developed primary conjunctival melanoma, histologically characterised by tumour thicknesses of 4, 4, 1.1, 3, and 2mm. Two or more conjunctival melanomas manifested ulceration, significant mitotic rates, necrosis, angiotropism, and intralesional transformation. The conjunctival melanoma recurred in a matter of months as one or more discrete satellite in transit lesions in the vicinity of the primary melanoma. Histological examination revealed well-defined micronodules containing atypical melanocytes in the subepithelial connective tissue stroma. All lesions were extravascular and most appeared angiotropic. Four of five patients subsequently developed parotid or other loco-regional nodal disease and rapidly ensuing widespread metastases and death. The time course from diagnosis to the demise of the patients averaged about 13 (range 7-20) months. Our findings suggest that satellite in transit metastases constitute an important new risk marker for possible rapid metastatic disease progression and death in patients with conjunctival melanoma. This finding appears to take on even greater significance if such lesions develop rapidly, i.e., in a matter of weeks or months following diagnosis of primary conjunctival melanoma, and if the primary melanoma manifests additional high-risk features. Additional studies are underway in order to further elucidate the mechanism of these metastases.

A Retrospective Review of Conjunctival Melanoma Presentation, Treatment, and Outcome and an Investigation of Features Associated With BRAF Mutations.

Large studies investigating clinical presentation and treatment in primary conjunctival melanoma (CM) are rare. Clinicopathological characteristics of BRAF-mutated CM have not been studied thoroughly. To determine the associations of clinicopathological tumor features and treatment with local recurrence, metastasis, and mortality and to determine the association of BRAF mutations with these features. Population-based cohort study at the Eye Pathology Institute, Copenhagen, Denmark. Participants included 139 patients with primary CM in Denmark from January 1, 1960, to December 31, 2012. For BRAF analysis, all patients with available formalin-fixed, paraffin-embedded tumor samples from January 1, 1994, to December 31, 2012, were included. BRAF mutations, local recurrence, regional and distant metastasis, melanoma-related mortality, and all-cause mortality were examined. A poor prognosis of tumors involving the extrabulbar conjunctiva and adjacent tissue structures was confirmed in multivariable Cox proportional hazards regression models. Patients undergoing incisional biopsy more frequently developed metastasis (hazard ratio [HR], 2.46; 95% CI, 1.08-5.58; P = .03). Excision without adjuvant treatment was associated with local recurrence (HR, 1.97; 95% CI, 0.11-3.48; P = .02), metastatic disease (HR, 2.51; 95% CI, 1.07-5.91; P = .03), and all-cause mortality (HR, 1.80; 95% CI, 1.05-3.08; P = .03). BRAF mutations were identified in 19 of 47 primary CMs (40.4%) and were more frequent in younger patients (P = .005), less frequent in the extrabulbar conjunctiva (P = .05), more frequently classified as T1 tumors (P = .03), and rarely manifested with primary acquired melanosis (P = .001) or with a uniformly pigmented lesion (P = .006). Distant metastases developed in 6 of 19 BRAF-mutated CMs (31.6%) as opposed to 1 of 28 BRAF wild-type CMs (3.6%). No definitive association with distant metastasis was seen in multivariable Cox proportional hazards regression models. Incisional biopsy and excision without adjuvant therapy were associated with a poor outcome in patients with CM. Extrabulbar location was also associated with a poor outcome in multivariable analysis. BRAF-mutated CMs were frequent in younger patients and were rare in tumors involving the extrabulbar conjunctiva. Despite a more favorable location, BRAF-mutated tumors may be associated with more frequent distant metastasis.

HSP90 inhibitors and conjunctival melanoma

AbstractPurpose To investigate the expression of HSP90 in human conjunctival melanoma (CM) cell lines and primary human melanocytes, and to assess the effects of HSP90 inhibitors on cell growth and proliferation.Methods CM cell lines (CRMM1 and 2; CM2005.1) and primary human melanocytes (n=3, P2 to P4) were immunolabelled for HSP90, and visualised using immunofluorescence and confocal microscopy. HSP90 expression was also examined in cell lysates from CM cell lines (CRMM1 and 2; CM2005.1) and primary melanocytes (n=3). The effect of several HSP90 inhibitors (17DMAG, SM252 and SM122) on cell growth and viability was assessed at 24hrs.Results CRMM1 and 2, and CM2005.1 cells showed strong cytoplasmic expression of HSP90, with low levels of cytoplasmic expression in primary melanocytes. This was confirmed using western blot. The HSP90 inhibitors induced dose‐dependent cell death in CM cells, 17DMAG (water soluble) being most effective (LD50 at 24 hrs ~30uM compared with ~50um for SM122 and SM122).Primary human melanocytes viability was not significantly reduced by the HSP90 inhibitors.Conclusion HSP90 is a widely expressed chaperone protein that can regulate cell growth, survival and angiogenesis pathways. CM cell lines expressed high levels of HSP90 protein compared to melanocytes, consistent with a recent study of primary conjunctival melanoma specimens, that found over‐expressed HSP90 in CM, particularly recurrent tumours. HSP90 inhibitors may be useful in inhibiting CM growth alone or in combination therapies. Funding: National Foundation for Medical Research &amp; Innovation (MCM &amp; RMC); Lions NSW Eyebank

C-Kit SCF receptor (CD117) expression andKITgene mutation in conjunctival pigmented lesions

To investigate the presence of KIT gene mutations and immunoreactivity in 85 conjunctival melanocytic tumours and to clarify the role of KIT as a potential therapeutic target in this group of patients. Eighty-five conjunctival pigmented tumours [27 melanomas, 12 primary acquired melanosis (PAMs) and 46 nevi] were immunostained for KIT. Intensity and pattern of expression were evaluated. Molecular analysis to identify KIT mutations was performed in 15 selected cases (tumour-rich areas >50%). KIT immunostaining score and pattern were statistically related to patients' age, sex, diagnostic category, presence of relapse, disease-free survival, presence of metastases, metastasis-free survival, limbal versus nonlimbal tumour location and thickness of melanomas. KIT stains were documented in 48% of melanomas, 50% of PAMs and 24% of nevi. The mean score of KIT staining in the melanomas/PAMs group was significantly different from nevi (p = 0.0076). No statistically significant differences were detected between either c-kit immunostaining score or pattern and each of the other clinico-pathologic parameters considered. No KIT gene mutations were detected in melanomas and nevi. A silent mutation/polymorphism in KIT exon 13 was found in one PAM. Despite the high level of KIT immunostains in PAMs and melanomas, this parameter seems not to be a good predictor of the presence of molecular mutations. KIT-activating mutations should be considered an uncommon event in this tumour.

Therapy of melanocytic conjunctival tumors

Clinical experience of our single center in dealing with pigmented epibulbar lesions - melanocytic conjunctival tumors is presented. Since 2008 we use the topical treatment with mitomycin C (MMC) as an alternative or adjunctive method for excision in the treatment of melanocytic neoplasia of the conjunctiva. The retrospective case series of 85 patients with pigmented lesions of the conjunctiva - melanocytic conjunctival tumors, histopathologically examined in the period 2001-2010 is presented. Since 2008 we started to apply MMC in the treatment of primary acquired melanosis (PAM) and dysplastic nevi. We apply MMC topically directly after an excision as 2-times five minutes application. In 85 patients with pigmented lesions of the conjunctiva histopathological findings after excision of the lesion showed in 68 (80 %) cases melanocytic nevocelullar nevus, out of which 55 cases were combined and 13 cases were junctional nevi. In 60 (80 %) cases of melanocytic nevi atypia was found in 25 patients (42 %), nevus without atypia was present in 35 cases (58 %). PAM with atypia was found in 16 patients (classified since 2000). During the period of application of MMC we diagnosed only one patient with primary conjunctival melanoma. There was no presence of relapse of the pigmented lesion either after primary excision or after excision with MMC. Resection of more than one quadrant of bulbar conjunctiva in patients with pigmented lesions of the conjunctiva in cases of conjunctival nevus with atypia and PAM with atypia combined with topical MMC chemotherapy is an alternative therapy for residual pigmented lesions. There was no presence of relapse of pigmentation in area of excision with or without using MMC during the surgery in patients with PAM. The number of our patients is not sufficient yet to draw a conclusion (Fig. 6, Ref. 21).

Local conjunctival metastases from primary conjunctival melanoma: clinico-pathological correlation and implications

Background/AimsA retrospective service evaluation identified seven patients who developed local conjunctival metastases (LCMs) of the conjunctiva after a primary diagnosis of conjunctival invasive melanoma. The study was conducted to identify...

RUNX2 expression in conjunctival melanocytic proliferations

AbstractPurpose Amplification of RUNX2 (6p21.2), a transcription factor involved in bone formation and in tumor progression of prostate and breast cancer, has previously been identified in 76% (16/21) of primary conjunctival melanoma and in 100% (4/4) of metastatic conjunctival melanoma. The aim of this study was to investigate RUNX2 expression at the protein level in a panel of conjunctival melanocytic proliferationsMethods Expression of RUNX2 was assessed by immunohistochemistry in 26 conjunctival naevi (14 compound naevi, 11 subepithelial naevi and 1 cellular blue naevus), 13 PAM (9 PAM with atypia) and 25 conjunctival melanoma. Statistical analysis was performed with JUMP 8,0 software. Expression was regarded as negative if less than 10% of the tumor cells expressed RUNX2. Examination was independently assessed by two observers.Results RUNX2 was found in 3 naevi (11,5 %), in one PAM with atypia (7,6%) and in 5 melanoma (20 %) without significant variation between the 3 groups. In the melanoma group, there was no correlation between RUNX2 expression and TNM category, depth of invasion, proliferation index (Ki67), local lymphatic invasion and the presence of metastasis. There was however a significant correlation between RUNX2 expression and death (p=0,0191).Conclusion RUNX2 is expressed in a limited number of conjunctival melanocytic proliferations without significant differential expression between benign and malignant lesions. The biological significance of RUNX2 in conjunctival melanoma tumor progression requires further functional investigations.

Expression of cancer-testis antigens (MAGE-A1, MAGE-A3/6, MAGE-A4, MAGE-C1 and NY-ESO-1) in primary human uveal and conjunctival melanoma

AimMetastatic disease in ocular melanoma remains untreatable, is associated with late detection and is resistant to conventional systemic therapies. Many tumours including cutaneous melanoma express specific cancer-testis (CT) antigens and...

Multiplex Ligation-Dependent Probe Amplification of Conjunctival Melanoma Reveals CommonBRAFV600E Gene Mutation and Gene Copy Number Changes

To determine the occurrence of BRAF V600E gene mutations and copy number changes of all autosome arms and genes known to be frequently altered in tumorigenesis in primary and metastatic conjunctival melanomas (CoMs). DNA (200 ng) was analyzed by three multiplex ligation-dependent probe amplification assays (P027 uveal melanoma, P036 human telomere, and P206 spitzoid melanoma). Eight of 16 primary tumor samples and 4 of 6 metastatic samples showed BRAF V600E gene mutations. CDKN1A and RUNX2 (both 6p21.2) were amplified in 11 and 16 of 21 primary CoMs, respectively. In metastatic CoMs, MLH1 (3p22.1) and TIMP2 (17q25.3) were frequently amplified, and MGMT (20q26.3) and ECHS1 (10q26.3) were frequently deleted. The BDH (3q), FLJ20265 (4p), OPRL1 (20q), and PAO (10q) genes, representing the telomeres of their respective chromosome arms in the P036 assay, were frequently amplified in metastatic CoMs. No statistically significant associations were identified between BRAF mutation or CDKN1A or RUNX2 amplification and sex, age, histologic cell type, or patient survival. No copy number changes were associated exclusively with metastatic CoMs. However, further investigation of the role of CDKN1A and RUNX2 in CoMs development and that of MLH1, TIMP2, MGMT, and ECHS1 in metastatic CoMs is warranted. Validation of the observed gene and chromosome arm copy number changes in a larger cohort of primary and metastatic CoMs is necessary to identify the patients at highest risk for CoMs metastasis.