PURPOSE:To investigate temporal trends in the incidence of posterior uveal melanoma in Scandinavia (Sweden, Denmark, and Norway) between 1960 and 2022 and explore potential associations with changes in tumor size and patient age at diagnosis. DESIGN:Retrospective, registry-based cohort study utilizing nationwide data from the 3 Scandinavian countries. PARTICIPANTS:All patients diagnosed with posterior uveal melanoma (choroid or ciliary body) in Sweden (1960-2022), Denmark (1960-2022), and Norway (1993-2022), totaling 10,154 cases. Iris melanomas were excluded. METHODS:Crude, age-standardized, and log-transformed age-standardized incidence rates were calculated using direct standardization (1970-1974 population). Linear regression was applied to assess trends in incidence, largest basal diameter (LBD), tumor thickness, and patient age at diagnosis. A multivariate linear regression model tested whether tumor size or patient age accounted for the time-dependent increase in incidence. P values were Bonferroni-adjusted for multiple comparisons. MAIN OUTCOME MEASURES:Crude, age-standardized, and log-transformed age-standardized incidence rates of posterior uveal melanoma. RESULTS:Crude, age-standardized, and log-transformed age-standardized incidence rates increased significantly in all 3 countries. Combined Scandinavian age-standardized rates rose from 5.2 to 6.8 per million inhabitants and year in the 1960s to 6.1-8.7 in the 2010s (slope 0.033; 95% CI: 0.022-0.044). Log-transformed rates showed a similar upward trend (slope 0.002; 95% CI: 0.002-0.003). Tumor size (diameter and thickness) decreased, while patient age at diagnosis increased over time. Multivariate analysis confirmed a time-dependent increase in incidence, independent of tumor size or patient age. CONCLUSIONS:Posterior uveal melanoma incidence in Scandinavia has increased since 1960. The linear trend is independent of changes in tumor size or patient age.

Introduction:The brachytherapy plaques used for uveal melanoma treatment are reused following sterilization processes. Here, we report patients with adverse reaction following a new cleaning procedure for brachytherapy plaques. Case Presentations:The brachytherapy plaque sewn on the outer eyewall would be removed following the delivery of calculated radiation dose. They are reused following standard calibration testing and sterilization processes. From September 2021 to January 2022, 6 patients treated with ruthenium-106 plaque brachytherapy presented with severe unilateral pain, eyelid swelling, and conjunctival injection with hemorrhage 1-2 days following plaque insertion. The surgeons observed a frosted appearance of the plaques in contrast to the normal shiny look during removal. The unusual severe postoperative reactions and the frosted appearance of plaques led to further investigations. A change in sterilization procedures with the use of LifeClean™ instead of PeraSafe™ was noticed. Ruthenium-106 brachytherapy plaques contain a uniformly distributed radioactive source covered by a thin silver shield. A test was performed by using two plates of pure silver that underwent its respective cleaning procedure using either PeraSafe™ or LifeClean™. Conclusion:The test results demonstrated formation of silver chloride with LifeClean™, while no such formation was demonstrated with PeraSafe™. We did not observe any new cases of severe postoperative reaction or frosting of plaques after changing back to PeraSafe™. Chlorine salt deposits probably contributed to severe inflammatory reaction of ocular surface.

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%).

Introduction: This report presents a case of an exceptionally delayed distant recurrence of a choroidal melanoma, occurring 4 decades after the enucleation of the affected eye. Case Presentation: In 1977, a 29-year-old man underwent enucleation for a choroidal melanoma. At the age of 68 years, he was diagnosed with advanced prostate cancer. Although the metastatic prostate cancer responded to treatment, a persistent lung lesion warranted further examination. A lung biopsy, somewhat surprisingly, confirmed the presence of melanoma metastasis, 4 decades after the enucleation. The cells were positive for Melan-A, while no BRAF mutation was identified. Two years later, new lesions appeared in the liver, and CT showed progression with multiple new sites. A liver biopsy revealed again melanoma recurrence, and its choroidal origin was verified by the presence of a GNA11 mutation. The patient underwent radiation therapy for the lung and liver lesions, followed by immunotherapy. However, the patient died 11 months after the recurrence in the liver. In this case report, the micrometastatic melanoma cells appear to have remained dormant for an extended period, before the patient’s treatment in 1977, but the reason for the late reactivation from the dormant state remains unclear. Conclusion: The recurrence of a choroidal melanoma is substantiated by the histopathological and molecular analyses, including the finding of a GNA11 mutation. This case exemplifies a remarkably delayed distant recurrence of a choroidal melanoma, which manifested clinically 40 years following enucleation.

PURPOSE:In a population-based cohort of 960 uveal melanoma (UM) patients, we describe patients with three additional malignancies, including one unique patient with four synchronous primary malignancies. METHOD:A descriptive presentation of the clinical course and outcome for UM patients with three additional primary malignancies. RESULTS:After more than 20 years of follow-up of the UM cohort, 11 patients (1.1%) were diagnosed with three additional primary malignancies before, simultaneously or after UM. Among these, one patient had four synchronous primary malignancies, detected during workup for a symptomatic UM. All diagnoses were treated during the following 4 months, firstly the breast cancer, thereafter, the lung and pancreatic cancers and finally the UM. The patient died 3 years later of abdominal carcinomatosis due to the pancreatic cancer. The family history and gene testing did not disclose any genetic predisposition for cancer. A comparison of the four synchronous tumours, morphologically and immunohistochemically, showed no similarities and the expression of antibodies was different. CONCLUSION:Patients with UM may be diagnosed with non-ocular additional primary cancers. Thus, a comprehensive workup is obligatory and a further follow-up of the UM patients seems necessary. The UM is not always the main problem.

We report a 46-year-old male patient with retinocytoma who presented at the age of 31 asymptomatically. An intraocular retinal mass was incidentally found in his right eye, when he underwent ophthalmological assessment for refractive surgery. This tumor consisted of a calcified sessile basis partially covered by a pedunculated salmon-pink growth. Initially, the tumor was diagnosed as a retinocytoma with clinical suspicion of malignant transformation into retinoblastoma and treated by four sessions of laser photocoagulation. Six and a half years later, the tumor relapsed, and he was treated with a Ruthenium plaque. Following brachytherapy, he had two episodes of right-sided vitreous hemorrhage that spontaneously cleared up, and the remaining finding in the vitreous cavity was interpreted as asteroid hyalosis. He underwent vitrectomy about five years following brachytherapy. The analysis of the vitreous material revealed the presence of inactive vitreous seeds composed of small round blue cells, compatible with a type 2 regression.

Intraocular medulloepithelioma is a rare embryonal tumor that is believed to arise from the epithelium of the medullary tube. We report a 37-year-old female with medulloepithelioma presented at the age of 17 with a one-month history of left-sided visual deterioration and visible iris lesion. Birth history and medical and family histories were insignificant. The left eye revealed a vascularized iris mass. Further examination revealed a grey-white ciliary body mass and a subluxated lens with best-corrected visual acuity (BCVA) of 0.5. The patient underwent partial lamellar corneo-sclerouvectomy. The histological and electron microscopic findings revealed medulloepithelioma. To reduce the risk of recurrence of the probable malignant tumor, she was treated with Ruthenium plaque therapy about six weeks following surgical removal. Pars plana vitrectomy and lensectomy with laser photocoagulation of the peripheral retina were performed at the removal of the brachytherapy plaque. She regained her BCVA of 1.0 in her left eye 3.5 months following pars plana vitrectomy. At 20-year follow-up, no tumor recurrence was seen and her BCVA remained 1.0.

PURPOSE:The purpose of this study is to explore the frequency of additional primary malignancies in uveal melanoma (UM) patients and cause-specific mortality, to help guide surveillance strategies after UM.METHODS:All patients diagnosed with UM at Oslo University Hospital during 1990-2017 were eligible for inclusion. Linkage to the Cancer Registry of Norway obtained information on additional malignancies and cause of death throughout 2019. UM patients were categorized according to timing of additional malignancy (prior/simultaneously or after UM) or no additional cancer, and by UM stage at diagnosis. Age-adjusted mortality rates were presented per 1000 person-years with 95% confidence intervals (CI).RESULTS:The study population included 960 UM patients: 77% were diagnosed in stage and I/II and 56% were men. Mean age at diagnosis was 63 years. Additional malignancies were observed in 152 patients prior/simultaneous to UM, and in 120 patients >1 year after UM. Overall, mortality per 1000 person-years was 3.5 (95% CI 3.1-3.9) for UM and 3.0 (2.6-3.4) for other causes. Lowest UM mortality [1.3 (0.60-2.1)] was seen in patients with a second malignancy after UM, regardless of stage. Highest UM mortality was seen for UM patients in stage III/IV, both without [16.1 (13.2-19.1)] and with any additional malignancy [16.9 (6.6-27.3)].CONCLUSION:Our results support that UM patients frequently have additional malignancies, both before and after UM. Low-UM mortality in patients with a primary malignancy after UM, might indicate less aggressive UM. The cumulative UM mortality flattens about 10 years after diagnosis and annual follow-up of patients for 10 years seems adequate.

Purpose: To evaluate arteriovenous malformations (AVM) with swept-source (SS) optical coherence tomography (OCT) angiography (OCTA) in iris racemose hemangioma and compare it with traditional intravenous iris fluorescein angiography (IVFA). Methods: A cross-sectional observational clinical study was conducted on patients with iris racemose hemangioma with the ZEISS PLEX Elite 9000 SS OCT & OCTA. Results: Three eyes of three patients were imaged. Iris racemose hemangiomas demonstrated a tortuous, well-defined, and continuous course of the AVM. The ZEISS PLEX Elite 9000 SS OCT & OCTA allowed for a detailed visualization of the ARM and was superior to IVFA in depicting small caliber, fine vessels. Conclusions: SS-OCTA may provide a dye-free, no-injection, cost-effective method comparable to spectral domain OCTA and IVFA for diagnosing and monitoring iris racemose hemangiomas for growth and vascularity.

Endophthalmitis caused by Phialemonium species are rare. We report a 68-year-old female with unilateral Phialemonium curvatum endophthalmitis. She presented with a two day history of right-sided floaters and reduced vision to finger counting, initially treated as iridocyclitis with topical and systemic steroids. Due to worsening, a diagnostic vitrectomy was performed. The culture from the vitreous cavity yielded Phialemonium curvatum growth. She was treated with amphotericin B intravitreally once and voriconazole tablets for three months with gain of Best Corrected Visual Acuity (BCVA) to 0.3. Six weeks following discontinuation of voriconazole, the patient developed right-sided eye pain and deterioration of the visual acuity. A second diagnostic vitrectomy was performed and once again the culture demonstrated growth of Phialemonium curvatum. She was treated with voriconazole for six months. Further, she developed eye pain and reduced vision in her right eye about 2 years following the second diagnostic vitrectomy. A third diagnostic vitrectomy was performed which was negative. Two weeks following the third diagnostic vitrectomy, the patient presented with retinal detachment and macular hole in the same eye. Retinal detachment persisted even following surgical repair with silicone insertion. The patient declined further surgeries. About ten years following initial presentation she developed intolerable right-sided eye pain and phthisis. The eye was eviscerated.

Juxtapapillary retinal capillary haemangiomas are rare benign vascular tumors located adjacent to the optic disc. They may occur sporadically or in association with von Hippel-Lindau disease. Treatment modalities for juxtapapillary retinal capillary haemangiomas vary from observation to aggressive vitreoretinal surgery. A combination of different treatment modalities is often tried with modest success. We report a patient diagnosed with right-sided juxtapapillary retinal capillary haemangioma at the age of 36 in an amblyopic eye previously treated with patching and surgery. In 2013, at the age of 45 he presented with increasing shadow in his right visual field. The ophthalmological examination revealed a right-sided subtotal serous retinal detachment with 6.2×5.6×3.5 mm juxtapapilllary retinal capillary haemangioma and visual acuity of hand movements. He underwent vitreoretinal surgery, radiation of juxtapapillary retinal capillary haemangioma with intraocular strontium applicator and insertion of silicone oil, all in one session. We describe a novel treatment, the intraocular radiation with strontium. This can be applied in complicated cases where vitrectomy is necessary.

PURPOSE: To determine the size at which choroidal melanomas can metastasize and to report the characteristics of small fatal choroidal melanomas (SFCM). DESIGN: Retrospective case series. METHODS: Ten ocular oncology services submitted 45 patients with a choroidal melanoma 3 mm or less in thickness and 9 mm or less in largest basal diameter (LBD), when treated, who developed metastases. RESULTS: Median tumor thickness was 2.4 mm (range, 1.0-3.0 mm) and LBD 7.3 mm (range, 3.0-9.0 mm). Of 14 (31%) tumors that were first observed, 12 grew a median of 0.5 mm (range, 0.1-1.2 mm) in thickness and 1.0 mm (range, 0-3.0 mm) in LBD within a median of 7 months; 3 were initially smaller than 3 mm in LBD. Number of risk factors for growth and metastasis was 0 for 4% of the tumors; 60% were over 2 mm in thickness, 63% had subretinal fluid, 84% caused symptoms, 57% had orange pigment, and 92% were within 3 mm of the disc. Local recurrence occurred in 8 of 31 eyes (26%) treated conservatively. Median metastasis-free survival was 4.5 years (range, 0.8-15.7 years). Kaplan-Meier estimate of metastasis developing was 15% (95% confidence interval [CI], 7-26), 51% (95% CI, 36-64) and 85% (95% CI, 71-92) by 2, 5, and 10 years, respectively. By the time of analysis, 37 patients had died of metastasis after a median of 7 months. CONCLUSIONS: Choroidal melanomas less than 3.0 mm in LBD are highly unlikely to metastasize. Risk factors of an SFCM are similar to those for all choroidal melanomas of similar size. (C) 2019 The Author(s). Published by Elsevier Inc.

Background: Early confirmation of the effect of brachytherapy for choroidal melanoma showing that tumour coverage is valuable. The irradiated retinal pigment epithelium (RPE) commonly develops atrophy. This study compares the fundus autofluorescence (AF) changes to the development of RPE atrophy following brachytherapy. Methods: Retrospective study of 19 patients treated with 106Ru and 2 with 125I plaques with either a 3- or 6-month follow-up period. Ultra-widefield (UW) composite colour and AF images were obtained with Optomap 200Tx and interpreted as complete, partial, or no RPE changes and complete or partial hyperautofluorescence, hypoautofluorescence, or isoautofluorescence. Results: At the 3-month follow-up, 9 of 13 patients (69%) (95% confidence interval [CI], 0.389–0.896) treated with 106Ru plaques developed complete homogenous hyperautofluorescence surrounding the tumour, but only 1 of 13 (8%) (95% CI, 0.004–0.379) developed complete RPE atrophy at the same time point. Six patients in the 106Ru plaque group had their first follow-up with UW imaging at 6 months. Four of them developed homogenous hyperautofluorescence and none developed complete RPE atrophy around the tumour. The 2 patients treated with 125I did not demonstrate any clear RPE or AF changes. Conclusion: The effect of 106Ru plaque treatment on fundus UW imaging is detected as homogenous and well-demarcated hyperautofluorescence before visible RPE atrophy.

PURPOSE:Uveal melanoma (UM) has a high propensity for metastatic spread, and approximately 40-50% of patients die of metastatic disease. Metastases can be found at the time of diagnosis but also several years after the tumor has been removed. The survival of disseminated cancer cells is known to be linked to anchorage independence, anoikis resistance, and an adaptive cellular metabolism. The cultivation of cancer cells as multicellular tumor spheroids (MCTS) by anchorage-independent growth enriches for a more aggressive phenotype. The present study examines the differential gene expression of adherent cell cultures, non-adherent MCTS cultures, and uncultured tumor biopsies from three patients with UM. We elucidate the biochemical differences between the culture conditions to find whether the culture of UM as non-adherent MCTS could be linked to an anchorage-independent and more aggressive phenotype, thus unravelling potential targets for treatment of UM dissemination. METHODS:The various culture conditions were evaluated with microarray analysis, quantitative reverse-transcription polymerase chain reaction (qRT-PCR), RNAscope, immunohistochemistry (IHC), and transmission electron microscopy (TEM) followed by gene expression bioinformatics. RESULTS:The MCTS cultures displayed traits associated with anoikis resistance demonstrated by ANGPTL4 upregulation, and a shift toward a lipogenic profile by upregulation of ACOT1 (lipid metabolism), FADS1 (biosynthesis of unsaturated fatty acids), SC4MOL, DHCR7, LSS (cholesterol biosynthesis), OSBPL9 (intracellular lipid receptor), and PLIN2 (lipid storage). Additionally, the present study shows marked upregulation of synovial sarcoma X breakpoint proteins (SSXs), transcriptional repressors related to the Polycomb group (PcG) proteins that modulate epigenetic silencing of genes. CONCLUSIONS:The MCTS cultures displayed traits associated with anoikis resistance, a metabolic shift toward a lipogenic profile, and upregulation of SSXs, related to the PcG proteins.

Approximately 50% of uveal melanoma patients develop metastases. We want to evaluate the effect of stricter criteria on our data from our previous study correlating survival and bone marrow (BM) micrometastasis results using our immunomagnetic separation (IMS) method. Mononuclear cell fractions (MNC) isolated from BM were examined for tumour cells and the patients were classified as BM positive (BM+) or BM negative (BM-). The study originally included 328 consecutive patients with uveal melanoma from 1997 to 2006. The cohort was limited to 217 patients when we introduced cyto- or histopathological verification of melanoma cells in the patient as a main new criterion for inclusion. Tumour cells were found in BM-samples in 38.7% (95% CI, 32-45) at enrolment. Until the latest work-up 43.8% (95% CI, 38-50) of patients had developed melanoma metastases. After a minimum follow-up time of 8.5 years, 60.4% (95% CI, 54-66) of patients had died. The causes were: melanoma metastases 69.5%, another type of cancer 5.4% and non-cancerous causes 19.5%. Overall median survival was shorter for the BM- patients (11.3 years) (95% CI, 10-12) compared to the BM+ (16.5 years) (95% CI, 12-14), p = 0.04, log rank test. All-cause mortality and specific melanoma mortality estimated after 12 year follow-up showed a highly significant difference comparing BM- and BM+, p = 0.010 and p = 0,017, respectively. IMS yields a high fraction of BM+ samples due to micrometastasis at diagnosis and these cells appear to have a positive prognostic impact strengthening our previous report. The late recurrences support the concept of tumour dormancy.

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.

Purpose: Our objective was to study survival rates with the bone marrow (BM) results in a cohort of uveal melanoma patients with long follow-up.Methods: Mononuclear cell fractions isolated from BM were examined for tumour cells using our immunomagnetic separation (IMS) method. The patients were classified as BM positive or BM negative. Clinical follow-up, histopathological findings, vital status and cause of death were registered.Results: The study included 328 consecutive patients with uveal melanoma from 1997 to 2006. Tumour cells were found in BM samples in 29% (95% CI, 25-34) at enrolment (96 cases). After a minimum follow-up time of 6 years, 156 (48%) (95% CI, 42-53) melanoma patients had died. The causes were as follows: melanoma metastases 92 (59%), another cancer 20 (13%) and non-cancer 44 (28%). Nine patients were still living with melanoma metastases. Until the latest work-up, 101 (31%) (95% CI, 26-36) patients had developed melanoma metastases. Cyto- or histopathological verification of the metastatic lesions was obtained in 85 cases (84%). In the group with melanoma metastases, 28 tested BM positive at study entry (28%) (95% CI, 19-38). In total, 39 of 101 with metastases tested positive at least once after a maximum of three tests (39%) (95% CI, 29-49). The overall median survival from the first BM test was shorter for the BM negative patients (9.5 years) compared with the BM positive (14.4 years), p = 0.02, log rank test.Conclusion: Ocular melanoma cells detected in BM seem to have a positive prognostic impact on survival in contrast to our original hypothesis.

PurposeOptic disc melanocytoma (ODM) is a benign tumour that usually occurs on or adjacent to the optic nerve head. The aim of the study was to evaluate fundus autofluorescence (FAF) imaging as a diagnostic tool in ODM.MethodsRetrospective comparative case series study of six patients with ODM and a comparing group of four patients with juxtapapillary choroidal nevus (JCN) and four with juxtapapillary uveal melanoma (JUM). Clinical examination was supplemented with ultrasound B-scan examination and spectral-domain optical coherence tomography. FAF images were obtained with the 532-nm laser (Optomap P200Tx) from all patients.ResultsClinical examination in the ODM group revealed a dome-shaped, darkly pigmented tumour on or adjacent to the optic disc in all patients, with a mean tumour basal dimension 1.4mm and mean tumour thickness by ultrasonography of 1.0mm. FAF revealed a totally hypofluorescent mass with sharply demarcated, feathery edges. No hyperfluorescent changes due to orange pigment or subretinal fluid were seen. In contrast, patients with JCN and JUM manifested focal hyperfluorescence as well as larger hyperfluorescent areas at the tumour and its borders.ConclusionFundus autofluorescence imaging is a non-invasive adjuvant tool in the differential diagnosis of ODM characterized by lack of hyperfluorescence compared to JCN and JUM.

Uveal melanoma (UM) is the most common intraocular malignancy and is fatal in approximately 50% of the cases. Cancer relapse can be seen years after treatment. Diverse mechanisms have been proposed to explain the late relapse of UM e.g cancer dormancy and the presence of cancers stem cells (CSCs). UM has the disadvantage of being difficult to culture, thus a limited number of cell lines are available. 3D sphere cultures of early passages offer a lower degree of clonal selection and perhaps a more accurate reflection of the primary tumours. The propagation of cells as spheres is believed to enrich for a CSC-like population. 2D cultures were expanded in RPMI 1640 with 10% FBS, while 3D sphere cultures were expanded in hESC + MEF for 7–10 days. We compared UM primary tumours fresh frozen vs. 2D culture and 3D sphere culture of the same donors by microarray, qRT-PCR, transmission electron microscopy (TEM), immunohistochemistry (IHC) and chromatin immunoprecipitation (ChIP). 3D sphere cultures induce signalling pathways that enhance survival and increase oncogenic activity. They show ultrastructural resemblance to in vivo UMs. The metabolic shift seen in 3D cultures could reflect the survival patterns of pre-metastatic cancer cells. The molecular signature of the 3D cultures reveals the presence of attractive targets for future epigenetic modulation or cancer cell vaccines.