The Pediatric Choroidal and Ciliary Body Melanoma Study: A Survey by the European Ophthalmic Oncology Group.

PURPOSE: To collect comprehensive data on choroidal and ciliary body melanoma (CCBM) in children and to validate hypotheses regarding pediatric CCBM: children younger than 18 years, males, and those without ciliary body involvement (CBI) have more favorable survival prognosis than young adults 18 to 24 years of age, females, and those with CBI. DESIGN: Retrospective, multicenter observational study. PARTICIPANTS: Two hundred ninety-nine patients from 24 ocular oncology centers, of whom 114 were children (median age, 15.1 years; range, 2.7-17.9 years) and 185 were young adults. METHODS: Data were entered through a secure website and were reviewed centrally. Survival was analyzed using Kaplan-Meier analysis and Cox proportional hazards regression. MAIN OUTCOME MEASURES: Proportion of females, tumor-node-metastasis (TNM) stage, cell type, and melanoma-related mortality. RESULTS: Cumulative frequency of having CCBM diagnosed increased steadily by 0.8% per year of age between 5 and 10 years of age and, after a 6-year transition period, by 8.8% per year from age 17 years onward. Of children and young adults, 57% and 63% were female, respectively, which exceeded the expected 51% among young adults. Cell type, known for 35% of tumors, and TNM stage (I in 22% and 21%, II in 49% and 52%, III in 30% and 28%, respectively) were comparable for children and young adults. Melanoma-related survival was 97% and 90% at 5 years and 92% and 80% at 10 years for children compared with young adults, respectively (P = 0.013). Males tended to have a more favorable survival than females among children (100% vs. 85% at 10 years; P = 0.058). Increasing TNM stage was associated with poorer survival (stages I, II, and III: 100% vs. 86% vs. 76%, respectively; P = 0.0011). By multivariate analysis, being a young adult (adjusted hazard rate [HR], 2.57), a higher TNM stage (HR, 2.88 and 8.38 for stages II and III, respectively), and female gender (HR, 2.38) independently predicted less favorable survival. Ciliary body involvement and cell type were not associated with survival. CONCLUSIONS: This study confirms that children with CCBM have a more favorable survival than young adults 18 to 25 years of age, adjusting for TNM stage and gender. The association between gender and survival varies between age groups.

Pathogenic Germline Variants in Uveal Melanoma Driver and BAP1-Associated Genes in Finnish Patients with Uveal Melanoma.

Uveal melanoma (UM) is a rare yet aggressive eye cancer causing over 50% mortality from metastasis. Familial UM, amounting to 1%-6% of patients in Finland and the United States, mostly lack identified genetic cause, while 8% show associations with other cancer syndromes. We searched novel genetic associations for predisposition to UM, additional to already studied BAP1 and MBD4, by using targeted amplicon sequencing of 19 genes associated with UM, BAP1, or renal cell carcinoma in 270 consecutively enrolled Finnish patients with UM. Key UM drivers GNAQ, GNA11, CYSLTR2, PLCB4, EIF1AX, and SF3B1 lacked pathogenic germline variants. One patient carried the pathogenic BRCA1 variant c.3626del p.(Leu1209*), and one harbored a novel truncating MET variant c.252C > G p.(Tyr84*), classified as likely pathogenic. FLCN and BRCA2, previously identified with pathogenic variants in patients with UM, did not have such variants in our cohort. Two patients were heterozygous for a pathogenic recessive BLM variant c.2824-2A > T. None of the carriers of identified variants had familial UM. We identified BRCA1 and MET as genes with pathogenic germline variants in Finnish UM patients, each with a frequency of 0.4% (95% confidence interval, 0-2).

The Pediatric and Young Adult Choroidal and Ciliary Body Melanoma Genetic Study, A Survey by the European Ophthalmic Oncology Group.

Purpose: To explore the genetic background of choroidal and ciliary body melanoma among children and young adults, with special focus on BAP1 germline variants in this age group. Methods: Patients under the age of 25 and with confirmed choroidal or ciliary body melanoma were included in this retrospective, multicenter observational study. Nuclear BAP1 immunopositivity was used to evaluate the presence of functional BAP1 in the tumor. Next-generation sequencing using Ion Torrent platform was used to determine pathogenic variants of BAP1, EIF1AX, SF3B1, GNAQ and GNA11 and chromosome 3 status in the tumor or in DNA extracted from blood or saliva. Survival was analyzed using Kaplan-Meier estimates. Results: The mean age at diagnosis was 17 years (range 5.0-24.8). A germline BAP1 pathogenic variant was identified in an 18-year-old patient, and a somatic variant, based mainly on immunohistochemistry, in 13 (42%) of 31 available specimens. One tumor had a somatic SF3B1 pathogenic variant. Disomy 3 and the absence of a BAP1 pathogenic variant in the tumor predicted the longest metastasis-free survival. Males showed longer metastasis-free survival than females (P = 0.018). Conclusions: We did not find a stronger-than-average BAP1 germline predisposition for choroidal and ciliary body melanoma among children and young adults compared to adults. Males had a more favorable survival and disomy 3, and the absence of a BAP1 mutation in the tumor tissue predicted the most favorable metastasis-free survival. A BAP1 germline pathogenic variant was identified in one patient (1%), and a somatic variant based mainly on immunohistochemistry in 13 (42%).

Pediatric ocular melanoma: a collaborative multicenter study and meta-analysis.

PURPOSE: To investigate clinical manifestations and prognoses in pediatric patients (≤12 years old) with ocular melanoma. METHODS: This was a retrospective, multicenter cohort study with individual participant data (IPD) meta-analysis pooling available published cases, and unpublished cases from an international collaboration of seven ocular oncology centers. RESULTS: There were 133 eyes of 133 pediatric patients with choroidal or ciliary body (n = 66 [50%]), iris (n = 33 [25%]), conjunctival (n = 26 [19%]), and eyelid (n = 8 [6%]) melanoma. Overall, the mean patient age at presentation was 7 years (median, 8; range, 1-12 years), with 63 males (49%). The mean age by tumor site was 6.50 ± 3.90, 7.44 ± 3.57, 9.12 ± 2.61, and 5.63 ± 2.38 years, for choroid/ciliary body, iris, conjunctiva, and eyelid melanoma, respectively (P = 0.001). Association with ocular melanocytosis was seen in 15%, 11%, 4%, and 0%, respectively (P = 0.01). Frequency of ocular melanoma family history did not vary by tumor site (7%, 17%, 9% and 12%, resp. [P = 0.26]). After mean follow-up of 74, 85, 50, and 105 months (P = 0.65), metastasis was seen in 12%, 9%, 19%, and 13% of choroid/ciliary body, iris, conjunctiva, and eyelid melanoma, respectively. Death was reported in 5%, 3%, 8%, and 0%, respectively, with survival analysis indicating higher mortality in choroidal/ciliary body and conjunctival melanoma patients. CONCLUSIONS: Ocular melanoma in the pediatric population is rare, with unique clinical features and outcomes. Iris melanoma accounts for about one-third of pediatric uveal melanoma cases.

Very late orbital recurrence of choroidal melanoma four decades post enucleation.

Orbital recurrence of malignant choroidal melanoma is quite uncommon, occurring in about 3% of patients undergoing enucleation for large tumors. Orbital recurrences after more than 10 years from enucleation are even rarer. In literature, only few reports described orbital recurrence that occurred between 10 and 40 years after primary diagnosis. Herein we report a very late choroidal melanoma recurrence, 40 years post enucleation, of a 52 year-old female who had undergone left enucleation at the age of 12. She presented to our clinic for consultation in 2017, complaining of difficulty to contain the prosthesis, as well as, presence of small pigmented palpable nodules inferiorly in her anophthalmic socket. The patient was not aware of the medical condition which had lead to the enucleation. We requested her medical reports and detailed history through her family. We realized after reviewing her charts 40 years back, that her enucleation was due to malignant choroidal melanoma (CMM). Full screening was performed. After magnetic resonance imaging (MRI), that showed the presence of nodular masses in the anophthalmic socket, an excisional biopsy was performed. Histopathology confirmed the diagnosis of CMM (epithelioid and spindle cell type), supporting the hypothesis that residual melanoma cells may remain clinically dormant for long periods, even for decades. A literature review was performed in order to review similar cases and to understand and discuss multiple factors, which may explain this extremely delayed recurrence. To the best of our knowledge, this is the third case to be reported in the literature.

Ultra-wide-field imaging assessment of small choroidal pigmented lesions using red and green colour channels.

BACKGROUND: Diagnosis of small choroidal melanoma is mainly based on tumour thickness, subretinal fluid, or lipofuscin pigment. Ultra-wide-field imaging (UWF) allows depiction of choroidal lesions through a red (RC) and a green channel (GC). Aim of the study was to determine the utility of this tool in the detection of small choroidal melanoma. METHODS: Retrospective cross-sectional study of patients with small choroidal pigmented lesions up to 3 mm in thickness. All patients underwent clinical and imaging assessment including UWF. Lesions were subcategorized based on thickness and lesion type. A qualitative assessment ensued using the red and green channels feature. RESULTS: A total of 152 patients were included. Melanotic naevi (76/152,50%) and small choroidal melanomas (55/152,36%) were the predominant types. Thickness was <1 mm in 30% (46/152), 1-2 mm in 46% (70/152) and 2-3 mm in 24% (36/152) of cases. Two distinct imaging patterns were noted: dark on RC/undetectable on GC and dark on RC/light on GC. In melanotic naevi the dark on RC/light on GC pattern was significantly associated with increased tumour thickness (p = 0.006) and the presence of lipofuscin (p < 0.001) suggesting a potential prognostic significance. In small melanomas such an association was not established. The majority of small melanomas manifested a dark on RC/undetectable on GC pattern despite the presence of subretinal fluid and lipofuscin. CONCLUSIONS: UWF imaging of choroidal pigmented tumours with red-green channels revealed two distinct patterns. The dark on RC/light on GC pattern was more common in suspicious melanotic naevi, but not in small melanomas. The use of red-green channels is not a reliable diagnostic tool in the early detection of small melanomas.

Very late recurrence of iris melanoma: 45 years after treatment.

Iris melanoma is a rare tumour that accounts for ~3-4% of all uveal melanoma cases. In the past, surgical resection was the sole treatement option, whereas nowadays, it has been replaced by brachytherapy as treatment of choice. Surgical resection is still in use in selected cases but combined with adjuvant radiotherapy in cases of incomplete resection. Herein, we report the case of a very late recurrence, 45 years after surgical resection. A literature review was performed to detect the latest recurrence reported to date and to understand the etiology behind very late recurrences. The role of multiple factors is discussed to explain this extremely delayed growth.

Iris extramedullary hematopoiesis in choroidal melanoma.

PURPOSE: Extramedullary hematopoiesis (EMH) usually occurs in patients with loss of bone marrow hematopoietic function, and in a vast majority of cases, it involves the liver, the spleen, or the lymph nodes. We report EMH in the iris of patients enucleated for choroidal melanoma (CM). METHODS: We report a series of three patients with CM, two treated with primary enucleation and one with secondary enucleation. RESULTS: Histopathology revealed EMH in the iris of all patients. CONCLUSION: EMH of the iris can be associated with CM in the absence of any hematological or systemic disorders. To the best of our knowledge, our work is the first report of this abnormal histopathological finding.

Comprehensive Study of the Clinical Phenotype of Germline BAP1 Variant-Carrying Families Worldwide.

Background: The BRCA1-associated protein-1 (BAP1) tumor predisposition syndrome (BAP1-TPDS) is a hereditary tumor syndrome caused by germline pathogenic variants in BAP1 encoding a tumor suppressor associated with uveal melanoma, mesothelioma, cutaneous melanoma, renal cell carcinoma, and cutaneous BAP1-inactivated melanocytic tumors. However, the full spectrum of tumors associated with the syndrome is yet to be determined. Improved understanding of the BAP1-TPDS is crucial for appropriate clinical management of BAP1 germline variant carriers and their families, including genetic counseling and surveillance for new tumors. Methods: We collated germline variant status, tumor diagnoses, and information on BAP1 immunohistochemistry or loss of somatic heterozygosity on 106 published and 75 unpublished BAP1 germline variant-positive families worldwide to better characterize the genotypes and phenotypes associated with the BAP1-TPDS. Tumor spectrum and ages of onset were compared between missense and null variants. All statistical tests were two-sided. Results: The 181 families carried 140 unique BAP1 germline variants. The collated data confirmed the core tumor spectrum associated with the BAP1-TPDS and showed that some families carrying missense variants can exhibit this phenotype. A variety of noncore BAP1-TPDS -associated tumors were found in families of variant carriers. Median ages of onset of core tumor types were lower in null than missense variant carriers for all tumors combined (P < .001), mesothelioma (P < .001), cutaneous melanoma (P < .001), and nonmelanoma skin cancer (P < .001). Conclusions: This analysis substantially increases the number of pathogenic BAP1 germline variants and refines the phenotype. It highlights the need for a curated registry of germline variant carriers for proper assessment of the clinical phenotype of the BAP1-TPDS and pathogenicity of new variants, thus guiding management of patients and informing areas requiring further research.

Conjunctival Melanoma during Pregnancy.

PURPOSE: To describe the clinical and histopathological features of a conjunctival melanoma (CM) during early pregnancy. PROCEDURES: A 37-year-old, 20-week pregnant primigravida was referred to the Sheffield Ocular Oncology Service with a rapidly growing lesion arising from the right superior conjunctival fornix, noted from the first trimester of pregnancy. This was associated with pain and bloody discharge. Incisional biopsy confirmed the clinical suspicion of invasive CM. She was treated by primary surgical excision and cryotherapy under local anaesthesia. RESULTS: Histology of the excised specimen showed an invasive malignant melanoma with surrounding in situ conjunctival changes arising from a naevus. The melanoma was 10.5 mm thick, focally necrotic, and had a mitotic count of 11/mm2 focally. The patient responded well to surgical treatment. She gave birth to a healthy boy, and the placenta showed no evidence of metastatic melanoma. There has been no recurrence or distant metastasis during 5 years of follow-up. CONCLUSION: CM during pregnancy is extremely rare. Because of possible transformation to malignant melanoma, we recommend close monitoring of females known to have pigmented conjunctival lesions of the conjunctiva during pregnancy.

KI-67 immunopositivity in choroidal and ciliary body melanoma with respect to nucleolar diameter and other prognostic factors.

PURPOSE: To study the association of cell proliferation in uveal melanoma with the mean of the 10 largest nucleoli (MLN), microvascular prognostic factors, and survival. METHODS: Population-based, retrospective cohort study of 167 choroidal and ciliary body melanomas enucleated from 1972 to 1981. Mouse monoclonal antibody Ki-67 (clone 7B11) was used to identify proliferating cells. MLN was measured from silver stained slides. Kaplan-Meier and Cox regression analysis were performed. RESULTS: The median number of Ki-67 immunopositive cells was 8 per tumor cross-section (range, 0-272) corresponding to a median of 0.30 cells/mm2. The percentage of immunopositive cells was 0.02% of all cells (range, 0-3.2). Tumors with epithelioid cells tended to have a higher percentage of immunopositive cells (p = 0.089). Ki-67 immunoreactivity, MLN, and presence of extravascular loops and networks were independent predictors of melanoma-related mortality, whereas Ki-67 immunoreactivity was not an independent predictor of mortality when presence of epithelioid cells was in the model. CONCLUSIONS: Taken together with previous data, the results of this population-based study support the theory that high cell proliferation index is associated with presence of epithelioid cells and that presence of a high cell proliferation index is associated with a higher risk of metastatic death independent of MLN, microvascular density, and presence extravascular loops and networks.

Nucleolar diameter and microvascular factors as independent predictors of mortality from malignant melanoma of the choroid and ciliary body.

PURPOSE: To determine whether nucleolar diameter and microvascular factors are independent predictors of mortality in malignant uveal melanoma of the choroid and ciliary body. METHOD: A population-based retrospective cohort study was conducted of melanoma-specific and all-cause mortality in 167 consecutive patients who had an eye enucleated because of choroidal and ciliary body melanoma from 1972 through 1981. The largest nucleoli were measured from digital photographs of silver-stained tumors along a central 5-mm-wide linear field parallel to the base of the tumor. The mean of the 10 largest nucleoli (MLN) was calculated. Microvascular loops and networks and microvascular density (MVD) were assessed. Kaplan-Meier and Cox regression analyses were performed. Associations between MLN and other variables were determined. RESULTS: The MLN could be determined in 126 (75%) melanomas. It ranged from 2.60 to 6.18 microm (median, 4.05). The association of large MLN with the presence of epithelioid cells (P = 0.017) and high MVD (P = 0.0053) was statistically significant. MLN was not significantly associated with tumor diameter and microvascular loops and networks. The 10-year melanoma-specific survival decreased with MLN (0.74, 0.60, and 0.42, arranged in tertiles; P = 0.0060), presence of loops and networks (P = 0.0001), and increasing MVD (P = 0.0001). By Cox regression, MLN was an independent predictor of survival, when adjusting in turn for presence of epithelioid cells, loops and networks, and MVD. In multivariate models with MVD, the independent prognostic information carried by MLN decreased, but the model as a whole was a better predictor of survival. The magnitude of this effect depended on whether MLN was modeled as a continuous or categorical variable. CONCLUSIONS: In this population-based data set, MLN and microvascular loops and networks were unrelated, independent predictors of survival. MLN and MVD were found to be partially interrelated. Multivariate models that included MVD in addition to MLN fitted better with observed melanoma-specific survival than models that excluded MVD.

Microvascular loops and networks in uveal melanoma.

Microvascular patterns--three-dimensional architectural arrangements of microvessels and extravascular matrix in uveal melanoma--were discovered when investigators were looking for histopathological features of sufficient size to be imaged clinically. Evidence that these patterns may be formed by tumour cells and that they may be able to conduct plasma and blood as well as discovery of similar elements in other cancers make them of general importance. Of nine different patterns described, closed microvascular loops and networks have been studied most extensively. When cell type, microvascular density and nucleolar size are controlled for, these two patterns independently predict time to metastasis. In addition to visualization in tumour specimens stained with periodic acid-Schiff reagent, they can often be visualized clinically on confocal indocyanine green angiography. The presence of networks is clinically associated with probability of growth of small uveal melanocytic tumours and with the rate of regression of uveal melanoma after brachytherapy. Networks are also associated with development of exudative retinal detachment from uveal melanoma. Histopathological studies show that loops and networks are less common in tumours enucleated after irradiation and that they are frequently repeated in metastases of uveal melanoma. Avenues for immediate future research include detailed elucidation of the histogenesis of microvascular patterns and determination of these patterns in metastatic melanoma to identify new histopathological characteristics for prognostication when clinical metastases have developed.

Uveal melanoma among Finnish children and young adults.

PURPOSE: To characterize uveal melanoma (UM) among children and young adults in a high-risk region for this cancer. METHODS: The medical records of consecutive patients <25 years of age with UM treated from 1962 to 2009 were retrospectively reviewed. The following data were collected: sex, tumor location and size, tumor node metastasis (TNM) stage, vision, and survival. We compared data with five previous series reporting 70 matching patients and combined them for meta-analysis of survival. RESULTS: Of 1,185 UM patients, 18 were eligible. UM frequency in patients <25 years of age was 1.3%; in those ≤20 years of age, 0.6%. Median follow-up time was longer than in the previous series combined (11.6 vs 5.4 years). Females outnumbered males 2:1. Median tumor thickness was higher in our series (8 vs 5 mm) and increased with age. Median tumor diameter was similar to previous series (12 mm). Of our patients, 83% were stage II; 17%, stage III. In previous series, 26% were stage I; 64%, stage II; and 10%, stage III. Survival was 76% at 5 and 10 years, compared to 98% in previous series. By meta-analysis, mortality increased with stage, age > 17 years, female sex, and if the ciliary body was involved. CONCLUSIONS: Tumor stage was higher than in other regions. Age >17 years, female sex, and tumor stage adversely influenced survival among patients <25 years of age with UM.

Prognostic associations of insulin-like growth factor-1 receptor in primary uveal melanoma.

OBJECTIVE: To study the association between immunoreactivity for insulin-like growth factor-1 receptor (IGF-1R) in primary ciliary body and choroidal melanoma and metastatic death in a consecutive, population-based data set. DESIGN: Retrospective, consecutive, population-based cohort study. PARTICIPANTS: A total of 167 patients with choroidal and ciliary body melanoma, enucleated from 1972 to 1981, with long-term survival data. METHODS: Specimens were immunostained by using the avidin-biotinylated peroxidase complex method and polyclonal antibodies to IGF-1R. The percentage of tumour area that was immunopositive was recorded. Survival was assessed by Cox multivariate regression analysis. RESULTS: The tumour area could be reliably measured from 129 (78%) of the 167 choroidal or ciliary body melanomas. More heavy pigmentation (p = 0.001), larger number of macrophages (p = 0.003) and higher microvascular density (p = 0.060) were associated with a higher percentage of tumour area that was immunopositive for IGF-1R, the reverse being true of extrascleral extension (p = 0.049). No significant association was observed with ciliary body extension, largest basal tumour diameter, cell type, mean diameter of the 10 largest nucleoli, and presence of extravascular matrix loops and networks (p = 0.61-0.96). The percentage of tumour area that was immunopositive for IGF-1R was not associated with survival. CONCLUSIONS: In our data set, immunoreactivity for IGF-IR did not independently predict metastasis from primary uveal melanoma. Partial loss of antigenicity can not be ruled out as a confounding factor because no frozen sections were available. Results of previous studies have likewise been variable, suggesting that immunohistochemical determination of IGF-1R from formalin-fixed, paraffin-embedded specimens is not practical as a routine test.

Nucleolar size in choroidal and ciliary body melanomas and corresponding hepatic metastases.

PURPOSE: This study aimed to investigate the relationship between hepatic metastasis and the mean diameter of the 10 largest nucleoli (MLN) in uveal melanoma. METHODS: A cross-sectional histopathological analysis of 37 metastases (13 surgical or needle biopsies, 24 autopsies) and corresponding primary choroidal and ciliary body melanomas was conducted, using statistical tests appropriate for paired data. The largest nucleoli were measured from digital photographs of silver-stained sections along a 5-mm-wide linear field. Confounders considered were presence of epithelioid cells and microvascular density (MVD), counted as the number of discrete elements labelled by monoclonal antibody QBEND/10 to the CD34 epitope. RESULTS: Hepatic metastases had more frequent epithelioid cells (p = 0.0047) and a higher MVD (median difference, 7.5 counts/0.313 mm(2) more; p = 0.044) than their corresponding primary tumours. Hepatic metastases, especially in autopsy specimens rather than surgical biopsies, tended to have a smaller MLN (median 3.6 mum) than the corresponding primary tumour (median difference, 0.55 mum; p = 0.066). The MLN in hepatic metastases was not associated with presence of epithelioid cells and MVD. Overall survival after diagnosis of metastasis was comparable whether hepatic metastases had a large or small MLN (p = 0.95), whereas a high MVD tended to be associated with shorter survival (p = 0.096) among the 13 patients with known survival. CONCLUSIONS: The results suggest that MLN is not a useful marker for assessing prognosis after diagnosis of hepatic metastasis from uveal melanoma.