Metastatic Cutaneous Melanoma Research Articles

Pembrolizumab-induced secondary adrenal insufficiency and thyroid dysfunction in an 81 year old male with metastatic cutaneous melanoma - A Case Report

Pembrolizumab-induced secondary adrenal insufficiency and thyroid dysfunction in an 81 year old male with metastatic cutaneous melanoma - A Case Report

Uveal melanoma immunogenomics predict immunotherapy resistance and susceptibility

Immune checkpoint inhibition has shown success in treating metastatic cutaneous melanoma but has limited efficacy against metastatic uveal melanoma, a rare variant arising from the immune privileged eye. To better understand this resistance, we comprehensively profile 100 human uveal melanoma metastases using clinicogenomics, transcriptomics, and tumor infiltrating lymphocyte potency assessment. We find that over half of these metastases harbor tumor infiltrating lymphocytes with potent autologous tumor specificity, despite low mutational burden and resistance to prior immunotherapies. However, we observe strikingly low intratumoral T cell receptor clonality within the tumor microenvironment even after prior immunotherapies. To harness these quiescent tumor infiltrating lymphocytes, we develop a transcriptomic biomarker to enable in vivo identification and ex vivo liberation to counter their growth suppression. Finally, we demonstrate that adoptive transfer of these transcriptomically selected tumor infiltrating lymphocytes can promote tumor immunity in patients with metastatic uveal melanoma when other immunotherapies are incapable.

Combining Immunotherapy and Targeted Therapies With Cryotherapy in Patients With Cutaneous Metastatic Melanoma.

Combining Immunotherapy and Targeted Therapies With Cryotherapy in Patients With Cutaneous Metastatic Melanoma.

Abstract 2905: Spatial analysis of neuropilin 2 expression in the microenvironment of melanoma

Abstract Background: Neuropilin 2 (NRP2) is a co-receptor that enhances signaling of growth factor and cytokine receptors and is expressed in tumor and microenvironment (TME) cells in melanoma and other tumors. Neuropilin 2 expression in tumor cells is associated with more aggressive tumors. Expression in tumor associated macrophages enhances efferocytosis and immunologically silent tumor cell death. Methods: Whole excisional lymph node biopsies were obtained from 10 treatment naïve patients with metastatic cutaneous melanoma who subsequently received anti-PD1 treatment. A single 5mm formalin-fixed paraffin embedded tissue section was used to assess a panel of 39 analytes by cyclic multiplex immunofluorescence (MxIF). Fields of view (~1mm2, n=288) were selected from pathologist-annotated regions of the tumor core, tumor-immune interface and adjacent lymphoid tissue. Pixel-based single cell segmentation and a supervised classifier approach was applied to resolve 12 distinct tumor, stromal and immune cell phenotypes and functional states (NRP2 positive or negative) in 1.5 million cells. Results: Tumor recurrence was observed in 6 patients (relapsed), whereas 4 patients were recurrence-free (non-relapsed) during follow up. Among the different cell types, NRP2 expression was more frequent in macrophages (44%), tumor cells (40%), dendritic cells (40%), followed by Tregs (31%) and T helper cells (29%), and less frequent in neutrophils (24%), cytotoxic T cells (23%) and B cells (15%). In colocalization permutation analysis, NRP2 expressing macrophages clustered away from tumor cells in non-relapsed patients while a random spatial distribution of NRP2 expressing macrophages with respect to tumor cells was observed in relapsed patients. In comparing the odds of tumor cell interactions with NRP2-expressing macrophages to those with NRP2 non-expressing macrophages, we found in the tumor:stromal interface (10% tumor cells) that cells from both non-relapsed and relapsed patients were more likely to interact with NRP2 non-expressing macrophages (OR = 0.73, 95% CI = [0.71, 0.75] and OR = 0.56, 95% CI = [0.55, 0.58] for non-relapsed and relapsed patients, respectively). However, in the tumor core (90% tumor cells), tumor cells from relapsed patients had significantly higher odds of interacting with NRP2 expressing macrophages, (OR = 1.15, 95% CI = [1.12, 1.18]), and this was not seen for non-relapsed patients (OR = 0.68, 95% CI = [0.66, 0.71]). In FOVs dominated by tumor cells (75% tumor), tumor cells from relapsed patients had 4.97 times the odds of expressing NRP2 compared to tumor cells from non-relapsed patients (95% CI = [2.23, 11.09], p = 0.0001). Conclusions: These spatial analytics suggest complex interactions among NRP2 expression in tumor and TME of melanoma determining risk for relapse with anti-PD1 treatment. Citation Format: Anastasios Dimou, Raymond M. Moore, Caitlin Ward, Alexey Leontovich, Ruifeng Guo, Chen Wang, Shankar Suman, Betty Dicke, Jill M. Schimke, Noah A. Stueven, Chathu A. Atherton, Wendy Nevala, Svetomir N. Markovic. Spatial analysis of neuropilin 2 expression in the microenvironment of melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2905.

Abstract A005: Quantification of cancer effect sizes of somatic variants reveals stage-dependent evolution in primary and metastatic skin cutaneous melanoma

Abstract The identities of driver and passenger genes in skin cutaneous melanoma (SKCM) have been intensively studied. However, the relative enhancement of cancer cell lineage survival and proliferation by SKCM driver mutations, as well as how these cancer effects vary with cancer stage, remains limited. The cancer effect size (also known as the scaled selection coefficient) quantifies the relative cancer cell lineage survival and proliferation in the context of each driver mutation. Using a diverse set of 999 primary and 1149 metastatic SKCM samples from multiple datasets, including The Cancer Genome Atlas (TCGA) and the AACR Project GENIE, we computed the cancer effect size of all recurrent mutations in primary (Stage I–III) and metastatic (Stage IV) tumors. Our analysis revealed that both primary and metastatic tumors were alike in their trinucleotide mutation profiles. However, the underlying gene mutation rates typically increased from primary to metastatic tumors as the latter group typically features ineffective DNA repair machinery. Additionally, the two cohorts noticeably differed in both magnitudes of effect sizes and in which genes specific somatic variants were most highly selected. Effect sizes of several driver mutations, most notably BRAF V600E, were orders of magnitude higher in metastatic tumors than in primary tumors. Furthermore, certain variants, particularly NONO R184L, were highly selected along the trajectory from normal tissue to primary tumors, yet new mutations were no longer selected between primary tumor resection and the development of resectable metastases. Conversely, for several variants, including H3F3B R18L, TP53 R273L, and PIK3CA R115P, selection was very strong only along the trajectory from resectable primary tumors to metastases. These findings illustrate the shifting evolutionary trajectory of tumors across cancer stages, underscoring the importance of early detection of SKCM and informing the development of targeted therapeutics aimed at specific stages of tumorigenesis. Citation Format: Rishi M Shah, Moein Rajaei, Jeff Mandell, Jeffrey P Townsend. Quantification of cancer effect sizes of somatic variants reveals stage-dependent evolution in primary and metastatic skin cutaneous melanoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A005.

Survival of patients with unfavorable prognosis cutaneous melanoma with increased use of immunotherapy agents: apopulation-based study in Belgium.

Although metastatic cutaneous melanoma is associated with an unfavorable prognosis, innovative therapies including immunomodulating agents and targeted therapies have shown survival benefits in clinical trials. We assessed the impact of the introduction of innovative drugs into clinical practice on the survival of patients with metastatic cutaneous melanoma during the period 2004-2017, in Belgium. The evolution of associated expenses was also analyzed. This is a retrospective population-based study using data from the national Belgian Cancer Registry, compulsory health insurance, and administrative survival data. The immunomodulating drugs were ipilimumab, nivolumab and pembrolizumab, while targeted therapies included vemurafenib, dabrafenib and trametinib. We did not identify a trend for improvement over time. Median survival (years) was 1.5 (95% CI: 1.1-1.8) in 2004-2008, 1.1 (95% CI: 0.8-1.5) in 2009-2013, and 1.6 (95% CI: 1.3-2.4) in 2014-2017, respectively. In contrast, survival improved in those with unknown primary tumor localization. In this group, median survival time was 2.0 (95% CI: 1.4-2.9) in the most recent period, while it was 1.1 (95% CI: 0.7-1.3) in 2009-2013, and 0.9 (95% CI: 0.6-1.2) in 2004-2008. The uptake of innovative drugs remained modest, with no drug being used by more than 30% of patients. Yearly expenditure was almost non-existent, and gradually increased, reaching several million euros in 2014-2017. Patients with metastatic cutaneous melanoma who were diagnosed between 2004 and 2017 showed no apparent improvement in survival. In contrast, increased survival was observed in the subgroup of patients with unknown primary tumor localization.

Case Series of Metastatic Cutaneous Malignant Melanoma to the Gallbladder and the First Reported Case of Metachronous Adenocarcinoma of the Colon

Two female patients in their fifties with a previous history of cutaneous malignant melanoma were found during follow-up to have a 'hot' lesion in the gallbladder on a Positron Emission Tomography scan. Imaging showed a gallbladder polyp. Histology revealed infiltration of the polyp mucosa by metastatic malignant melanoma. One case had a BRAF mutation. A male in his 70s was found on a staging computed tomography scan to have a suspicious intraluminal lesion in the gallbladder and thickening of the sigmoid colon. Subsequent histology confirmed metastatic malignant melanoma in the chest wall and to the gallbladder and adenocarcinoma in the colon. Molecular testing showed BRAF mutation. The metachronous adenocarcinoma in the colon was mismatch repair protein proficient and had no KRAS mutation. Histology from all cases showed that metastatic malignant melanoma to the gallbladder is superficial. Discussion: Reports from autopsy examinations revealed that metastasis from malignant melanoma to the gallbladder can be up to 15% - 20%. Most patients have mild symptoms or are asymptomatic which explains the paucity of cases reported in living patients within the published literature. Most of the previous reports showed the metastatic malignant melanoma to the gallbladder presented macroscopically as a polyploidal lesion. Conclusion: Our histological observation and previous reports showed that metastatic malignant melanoma in the gallbladder tends to be superficial. All our cases show no lymphatic or vascular invasion in the histological examination as previously published reports, however, the hematological spread is the most commonly suggested mechanism of spread.

Real-World Outcomes of Inoperable and Metastatic Cutaneous Head and Neck Melanoma Patients.

This study aims to describe the overall survival (OS) and to identify associated prognostic factors in patients with inoperable and metastatic cutaneous melanoma of the head and neck (H&N) region, undergoing modern systemic treatments. This is a retrospective single institutional study. Data on all consecutive H&N melanoma patients treated with systemic oncologic treatments between 2015 and 2022 were collected from electronic medical files. Kaplan-Meier curves were used to describe survival and Cox regression analysis was used to identify patient and tumor factors associated with prognosis. A total of 144 patients were included. Median OS was 45 months (95% confidence interval [CI] 28-65 m). On univariable analysis for OS, the primary disease site, specifically the nape and neck (hazard ratio [HR] 3.3, 95% CI 1.4-7.7, p = 0.007), high Eastern Cooperative Oncology Group Performance Status ([ECOG-PS], HR 2.5, 95% CI = 1.9-3.3, p < 0.001), high lactate dehydrogenase (LDH) levels (HR 2.8, 95% CI = 1.7-4.6, p < 0.001), and treatment with targeted therapy (TT) as compared with immunotherapy (HR 2.6, 95% CI = 1.06-6.3, p = 0.03) were all associated with shorter OS. High-grade adverse events (AEs) were associated with a longer OS (HR 0.41, 95% CI = 0.25-0.68, p = 0.001). On multivariable analysis for OS, the ECOG-PS, LDH levels, site of disease, and the development of moderate-severe AEs remained significant. In the era of modern oncologic treatments, the prognosis of inoperable and metastatic cutaneous H&N melanoma aligns with other cutaneous melanomas. Primary tumor site of the nape and neck region emerges as a significant prognostic factor. 3 Laryngoscope, 2024.

Distinct patterns of proteostasis network gene expression are associated with different prognoses in melanoma patients

The proteostasis network (PN) is a collection of protein folding and degradation pathways that spans cellular compartments and acts to preserve the integrity of the proteome. The differential expression of PN genes is a hallmark of many cancers, and the inhibition of protein quality control factors is an effective way to slow cancer cell growth. However, little is known about how the expression of PN genes differs between patients and how this impacts survival outcomes. To address this, we applied unbiased hierarchical clustering to gene expression data obtained from primary and metastatic cutaneous melanoma (CM) samples and found that two distinct groups of individuals emerge across each sample type. These patient groups are distinguished by the differential expression of genes encoding ATP-dependent and ATP-independent chaperones, and proteasomal subunits. Differences in PN gene expression were associated with increased levels of the transcription factors, MEF2A, SP4, ZFX, CREB1 and ATF2, as well as markedly different survival outcomes. However, surprisingly, similar PN alterations in primary and metastatic samples were associated with discordant survival outcomes in patients. Our findings reveal that the expression of PN genes demarcates CM patients and highlights several new proteostasis sub-networks that could be targeted for more effective suppression of CM within specific individuals.

Optimizing diagnostic success for vitreoretinal lymphoma

Aims/Purpose: To determine the diagnostic yield and pathological entities in a retrospective cohort of diagnostic vitrectomy patients.Methods: A retrospective cohort of diagnostic vitrectomy samples submitted to a tertial referral hospital pathology laboratory was identified by database search. The specimen submission protocol for this cohort consisted of two components: (i) intraoperative preparation of a neat vitreous aspirate as well as submission of the casette washings in RPMI and (ii) laboratory triage of each sample for microbiology culture, flow cytometry, cytological assessment and cell block preparation.Results: A total of 133 diagnostic vitrectomy samples were submitted between November 2015 and January 2022 (mean age 74 years, 0.8 M:F). A diagnostic report resulted in 132 (99.2%) of cases. Benign diagnoses (infectious and non‐infectious vitritis, degenerative or reactive changes, haemorrhage) were found in 88 of the cases (66.1%), while 20 cases (15%) were malignant. Of the malignant diagnoses, 18 (13.5%) were vitreoretinal lymphoma (17 diffuse large B cells; 1 T cell) by cytology and flow cytometry. The other two malignant diagnoses consisted of metastatic adenocarcinoma and metastatic cutaneous melanoma. 24 cases (18%) were classified as atypical, consisting of a lymphocytosis without demonstrable clonality by flow cytometry.Conclusions: This intraoperative and laboratory protocol maximizes the sample for all diagnostic tests required to discern infectious, inflammatory and malignant causes of increased vitreous cellularity. Importantly, the protocol allows for immunophenotyping of vitreoretinal lymphoma by flow cytometry and immunohistochemistry to WHO classification standards, essential for treatment and surveillance decision‐making.

Diagnostic challenges of posterior scleritis in pembrolizumab treated patient for cutaneous melanoma: A case report

PurposeTo describe various clinical pictures of posterior scleritis following pembrolizumab therapy for metastatic cutaneous melanoma. MethodsDescription of the clinical case with data from patient presentation and recurrences, diagnostic examinations, treatment, and six-months follow up. Clinical presentation of previous reports of pembrolizumab related of eye inflammation are discussed. ResultsA 49-year-old patient underwent resection of metastatic melanoma of the skin with sentinel lymph nodes removal and pembrolizumab adjuvant therapy every three weeks. Nine months after pembrolizumab initiation the patient accessed ophthalmology emergency service complaining one week vision loss on her right eye associated with pain on eye movement. Ocular examination revealed right eye papillitis and posterior scleritis. Intravenous corticosteroid treatment was started associated with topical prednisolone drops, after which complete resolution of the subretinal fluid was seen. During the six months follow up one episode of posterior scleritis was observed in the contralateral eye and resolved after peribulbar corticosteroid injection and oral therapy and one recurrence of intermediate scleritis in the right eye which resolved after oral therapy. ConclusionPosterior and intermediate scleritis may be added to the spectrum of pembrolizumab related eye inflammation; in the case presented posterior scleritis occurred asymmetrically in both eyes suggesting chronic features, importance of steroid tapering and follow up.

Clinical and economic analysis of treatment sequences with prolgolimab and BRAF/MEK inhibitors in adult patients with metastatic or unresectable cutaneous melanoma

Objective: evaluation of the comparative pharmacoeconomic effectiveness of treatment sequences with prolgolimab as the first line and combination therapy with BRAF/MEK inhibitors as the second line versus a regimen with BRAF/MEK inhibitors as the first line and prolgolimab as the second line in adult patients with metastatic or unresectable cutaneous melanoma.Material and methods. A detailed Markov and decision tree model was developed to allocate patients with metastatic cutaneous melanoma (mCM) with BRAF gene mutation (BRAF+) to treatment with prolgolimab or to targeted therapy with BRAF/MEK inhibitors (“dabrafenib + trametinib”, or “vemurafenib + cobimetinib” combinations). The costs of BRAF+ mCM therapy and the number of life years gained (LYGs) depending on the treatment regimen were calculated using approximated overall survival (OS) and progression-free survival (PFS) curves taken from relevant publications.Results. The treatment sequence for BRAF+ mCM had a significant impact on patient treatment outcomes: the median OS for the “prolgolimab → BRAF/MEK inhibitors” regimen was 41 months, while for the “BRAF/MEK inhibitors → prolgolimab” regimen it was 26 months; the median PFS was 11.5 months for the sequence starting with prolgolimab and 12.2 months for the strategy starting with “dabrafenib + trametinib” combination. The number of LYGs for a therapy regimen starting with prolgolimab and a regimen starting with “dabrafenib + trametinib” combination when modeling in the 1st year of therapy was 0.92 and 0.94 years, and at a 5-year horizon it was 3.19 and 2.75 years, respectively. At the same time, the cost of 1 LYG with a strategy starting with prolgolimab was 156 thousand rubles (5%) lower than the strategy starting with “dabrafenib + trametinib” combination.Conclusion. The developed pharmacoeconomic research model facilitated a clinical and economic analysis of using prolgolimab compared to targeted therapy with BRAF/MEK inhibitors across four lines of therapy, closely reflecting real clinical practice in the treatment of BRAF+ mCM patients.

Prognostic and predictive biomarkers in melanoma

Biomarkers help to inform the clinical management of patients with melanoma. For patients with clinically localised primary melanoma, biomarkers can help to predict post-surgical outcome (including via the use of risk prediction tools), better select patients for sentinel lymph node biopsy, and tailor catch-all follow-up protocols to the individual. Systemic drug treatments, including immune checkpoint inhibitor (ICI) therapies and BRAF-targeted therapies, have radically improved the prognosis of metastatic (stage III and IV) cutaneous melanoma patients, and also shown benefit in the earlier setting of stage IIB/C primary melanoma. Unfortunately, a response is far from guaranteed. Here, we review clinically relevant, established, and emerging, prognostic, and predictive pathological biomarkers that refine clinical decision-making in primary and metastatic melanoma patients. Gene expression profile assays and nomograms are emerging tools for prognostication and sentinel lymph node risk prediction in primary melanoma patients. Biomarkers incorporated into clinical practice guidelines include BRAF V600 mutations for the use of targeted therapies in metastatic cutaneous melanoma, and the HLA-A∗02:01 allele for the use of a bispecific fusion protein in metastatic uveal melanoma. Several predictive biomarkers have been proposed for ICI therapies but have not been incorporated into Australian clinical practice guidelines. Further research, validation, and assessment of clinical utility is required before more prognostic and predictive biomarkers are fluidly integrated into routine care.

Abstract A035: Comparing the clinical and genomic landscapes of acral, mucosal, and cutaneous metastatic melanomas treated with immune checkpoint inhibitors

Abstract Background: Melanoma is a melanocytic malignancy that is classified into different subtypes, including cutaneous (CM), acral (AM), and mucosal melanoma (MM). Immune checkpoint inhibitors (ICI) targeting PD-1 and/or CTLA-4 have emerged as the standard of care for advanced metastatic melanoma. We investigated responses to ICIs, genomic profiles, and molecular differences among these different subtypes of melanoma. Methods: We performed a multi-center retrospective cohort study including patients with metastatic melanoma who received anti-PD1 +/- CTLA4 inhibitor ICI for metastatic disease. We employed the AACR GENIE (v13.1) cancer database, DepMap portal, and Enrichr web tool with MSigDB Hallmark 2020 database to analyze the incidence and distribution of significant alterations, differentially expressed genes, and pathway enrichment within distinct melanoma subtypes. Results: We identified 337 patients with advanced melanoma who received anti-PD1 +/- anti-CTLA4 for metastatic disease. CM was the most frequent melanoma subtype (81%), followed by MM (12%), and AM (8%). Patients with AM had the shortest OS and PFS CM (OS 3.4 years, PFS of 1.1 years), AM median (OS 1.4 years, PFS 3.8 months), and MM (OS 1.7 years, PFS 6 months; OS, P= 0.028; PFS P=0.029). Patients with CM or MM experienced longer OS with anti-PD1 +/- CTLA4 vs. anti-PD1 monotherapy, but no survival advantage was observed in patients with AM. In the GENIE dataset, we identified 2015/374/177 samples with CM/MM/AM, respectively. AM and MM patients were significantly more likely to be Female, Black or Asian than CM patients. BRAF V600 mutations were most frequent in CM (40%), followed by MM (7%) and AM (14%). However, MM and AM had an increase in alterations in cell cycle regulator genes. The incidence of CDK4 and CCND1 amplification, respectively was highest in AM (17%, 16%), followed by MM (6%, 6%) and CM (2%, 3%). We compared the pathways represented by differentially altered genes in AM vs. CM: G2M-Checkpoint (Q&amp;lt;0.05) and E2F targets (Q&amp;lt;0.05) pathways were enriched in AM. We also compared RNAseq data from n=4) AM and n=20 CM cell lines, and found the same pathways enriched amongst genes that were differentially expressed between AM and CM: G2M-Checkpoint (Q&amp;lt;0.01) and E2F targets (Q&amp;lt;0.01) Conclusions: Our findings highlight comparatively poor outcomes with ICIs in Acral Melanoma. AM has lower rates of actionable BRAF mutations; as such, very few patients with metastatic AM have access to effective 1st or 2nd line therapies. Thus, there is an urgent need to explore alternative therapeutic targets. Our data implicate cell cycle inhibitors as potentially important components of novel treatment strategies that could augment immunotherapy efficacy for patients with metastatic AM. Citation Format: Sadaf Solati, April A. N. Rose, Anna Spreafico, Adrian Sacher, Denis Yahiaoui, Wilson H. Miller. Comparing the clinical and genomic landscapes of acral, mucosal, and cutaneous metastatic melanomas treated with immune checkpoint inhibitors [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A035.

Recent Advances in the Field of Artificial Intelligence for Precision Medicine in Patients with a Diagnosis of Metastatic Cutaneous Melanoma.

Standard-of-care medical imaging techniques such as CT, MRI, and PET play a critical role in managing patients diagnosed with metastatic cutaneous melanoma. Advancements in artificial intelligence (AI) techniques, such as radiomics, machine learning, and deep learning, could revolutionize the use of medical imaging by enhancing individualized image-guided precision medicine approaches. In the present article, we will decipher how AI/radiomics could mine information from medical images, such as tumor volume, heterogeneity, and shape, to provide insights into cancer biology that can be leveraged by clinicians to improve patient care both in the clinic and in clinical trials. More specifically, we will detail the potential role of AI in enhancing detection/diagnosis, staging, treatment planning, treatment delivery, response assessment, treatment toxicity assessment, and monitoring of patients diagnosed with metastatic cutaneous melanoma. Finally, we will explore how these proof-of-concept results can be translated from bench to bedside by describing how the implementation of AI techniques can be standardized for routine adoption in clinical settings worldwide to predict outcomes with great accuracy, reproducibility, and generalizability in patients diagnosed with metastatic cutaneous melanoma.

Endothelial and tumor-intrinsic mechanisms of hepatic melanoma metastasis.

The treatment of metastatic cutaneous melanoma was fundamentally improved by the discovery and introduction of immune checkpoint inhibitors, such as anti-PD-1 and anti-CTLA-4 antibodies, and targeted therapy with BRAF and MEK inhibition. Unfortunately, many patients suffer a relapse due to resistance mechanisms that in part are mediated by organ-specific metastatic sites. Especially, brain and liver metastases are negative predictive factors for both treatment modalities. There is still high unmet clinical need to prevent and treat spread to these organs. Therefore, experimental research should focus on mechanisms of hepatic melanoma metastasis to better understand this process and to identify therapeutic targets.

Caffeine reduces viability, induces apoptosis, inhibits migration and modulates the CD39/CD73 axis in metastatic cutaneous melanoma cells.

We aimed to evaluate the effect of caffeine on viability, apoptosis, migration, redox profile and modulatory effect of the purinergic system of cutaneous melanoma cells. The melanoma cells SK-MEL-28 and non-tumoural CCD-1059sk cells were treated for 24 h with different concentrations of caffeine. Cell viability was evaluated by a biochemical assay and fluorescence microscopy, and flow cytometry assessed apoptosis induction. A wound-healing assay assessed cell migration. The redox profile was evaluated by the levels of markers of reactive oxygen species (ROS), nitric oxide (NOx), total thiols (PSH) and non-protein thiols (NPSH). RT-qPCR and flow cytometry assessed the expression of CD39 and CD73. ATPase/ADPase and AMPase enzyme activities were evaluated by hydrolysis of ATP, ADP and AMP nucleotides. A bioluminescent assay assessed extracellular ATP levels. Caffeine significantly reduced melanoma cell viability and migration and did not affect non-tumoural cells. Caffeine increased ROS levels and improved PSH levels in melanoma cells. Furthermore, caffeine reduced CD39 and CD73 expression, decreased ATP, ADP and AMP nucleotide hydrolysis and increased extracellular ATP levels. We have shown that caffeine reduces metastatic cutaneous melanoma cell viability and migration, induces ROS generation and improves PSH levels. In an unprecedented manner, we also showed that caffeine reduces the expression of CD39 and CD73 and, consequently, ATPase/ADPase/AMPase hydrolytic activity of ectonucleotidases, thus displacing the CD39/CD73 axis and increasing extracellular ATP levels. Therefore, caffeine may be an interesting compound for clinical trials with the CD39/CD73 axis as a therapeutic target.

Predicting cutaneous malignant melanoma patients' survival using deep learning: a retrospective cohort study.

Cutaneous malignant melanoma (CMM) has the worst prognosis among skin cancers, especially metastatic CMM. Predicting its prognosis accurately could direct clinical decisions. The Surveillance, Epidemiology, and End Results database was screened to collect CMM patients' data. According to diagnosed time, patients were subdivided into three cohorts, train cohort (diagnosed between 2010 and 2013), validation cohort (diagnosed in 2014), and test cohort (diagnosed in 2015). Train cohort was used to train deep learning survival model for cutaneous malignant melanoma (DeepCMM). DeepCMM was then evaluated in train cohort and validation cohort internally, and validated in test cohort externally. DeepCMM showed 0.8270 (95% CI, confidence interval, CI 0.8260-0.8280) as area under the receiver operating characteristic curve (AUC) in train cohort, 0.8274 (95% CI 0.8286-0.8298) AUC in validation cohort, and 0.8303 (95% CI 0.8289-0.8316) AUC in test cohort. Then DeepCMM was packaged into a Windows 64-bit software for doctors to use. Deep learning survival model for cutaneous malignant melanoma (DeepCMM) can offer a reliable prediction on cutaneous malignant melanoma patients' overall survival.

RESOLUTION OF CHOROIDAL METASTASIS AND ASSOCIATED SUBRETINAL EXUDATION OF METASTATIC CUTANEOUS MELANOMA 15 DAYS AFTER TREATMENT WITH COMBINED TARGETED THERAPY.

To report a case of metastatic cutaneous melanoma to the choroid with rapid and complete resolution of associated choroidal elevation and subretinal fluid after initiation of combined targeted therapy. We describe a case of a 41-year-old man diagnosed with a metastatic cutaneous melanoma to the choroid of his right eye, for which treatment with dabrafenib/trametinib was initiated. A 41-year-old man with a past medical history of a BRAF V600E/V600E2/V600D mutated invasive superficial spreading cutaneous melanoma presented with acute metamorphopsia and blurred vision in his right eye. Examination revealed a best-corrected visual acuity (BCVA) of 20/22 and 2 elevated choroidal lesions temporal to the fovea with subretinal exudation to the fovea on fundoscopy. On repeat examination 3 days later, his vision had further decreased to 20/50 with an increase of subretinal fluid. Treatment with BRAF/MEK-inhibitor dabrafenib/trametinib was initiated, with complete resolution of the choroidal masses and subretinal exudation and improvement of the BCVA to 20/22 after only 15 days. Follow-up 8 weeks after start of therapy showed stable fundoscopic and tomographic findings, with further improvement of BCVA to 20/17 and no ocular side effects. A case of metastatic cutaneous melanoma to the choroid with choroidal elevation and subretinal exudation to the fovea, for which treatment with dabrafenib/trametinib was initiated. Rapid and complete resolution of choroidal metastasis and the associated subretinal exudation after initiation of combined targeted therapy was seen, without any ocular side effects.

Comprehensive in silico analysis of glycosylphosphatidylinositol- anchored protein (GPI-AP) related genes expression profiles in human normal and cancer tissues.

In human cells, there are more than 146 glycosylphosphatidylinositol-anchored proteins (GPI-APs), including receptors, ligands, adhesion molecules and enzymes. The proteins are associated with membrane microdomains called lipid rafts through GPI, and plays a variety of important biological functions. At present, plenty of studies have been carried out on the biosynthesis of GPI-APs. The biosynthesis of GPI-APs requires at least 20 steps, and more than 40 GPI biosynthetic genes have been identified. However, it remains unclear how expression of GPI-AP related genes is regulated in normal and cancer tissues. In this study, we utilized gene expression data from both the TCGA database and GTEx portal to analysis the gene expression involved in GPI-AP biosynthesis and encoding GPI-APs in normal and cancer tissues. In order to perform a comprehensive analysis, we employed the GlycoMaple, a tool that is specifically designed to analyze glycosylation pathways. The results showed that compared with normal tissues, the expression of genes involved in GPI-AP biosynthesis in cancer tissues such as early glioma, glioblastoma multiforme, pancreatic cancer, testicular germ cell carcinoma, skin primary cutaneous melanoma and skin metastatic cutaneous melanoma, was changed significantly. Particularly, the expression of PIGY in these six cancers was increased. In addition, the expression of CD14, a GPI-AP gene, was increased in these six cancers. The expression of GAS1, GPC2 and GPC4 was increased only in early glioma and glioblastoma multiforme indicating that some GPI-APs such as GAS1 can be used as biomarkers of glioma. This study provides new insights into the expression of GPI-AP related genes in normal and cancer tissues, and lays a solid foundation for the development of GPI-APs as biomarkers.