Molecular Profile of Subungual Melanoma: A MelaNostrum Consortium Study of 68 Cases Reporting BRAF, NRAS, KIT, and TERT Promoter Status.

BACKGROUND: Subungual melanoma (SM) is an unusual type of melanocytic tumor affecting the nail apparatus. The mutational prevalence of the most prominently mutated genes in melanoma has been reported in small cohorts of SM, with unclear conclusions on whether SM is different from the rest of melanomas arising in acral locations or not. Hence, the molecular profile of a large series of SM is yet to be described. OBJECTIVES: The aim of this study was to describe the molecular characteristics of a large series of SM and their association with demographic and histopathological features. METHODS: Patients diagnosed with SM between 2001 and 2021 were identified from six Spanish and Italian healthcare centers. The mutational status for BRAF, NRAS, KIT, and the promoter region of TERT (TERTp) were determined either by Sanger sequencing or next-generation sequencing. Clinical data were retrieved from the hospital databases to elucidate potential associations. RESULTS: A total of 68 SM cases were included. Mutations were most common in BRAF (10.3%) and KIT (10%), followed by NRAS (7.6%), and TERTp (3.8%). Their prevalence was similar to that of non-subungual acral melanoma but higher in SM located on the hand than on the foot. CONCLUSIONS: To date, this study represents the largest cohort of SM patients with data on the known driver gene mutations. The low mutation rate supports a different etiopathogenic mechanism for SM in comparison of non-acral cutaneous melanoma, particularly for SM of the foot.

Protocol to characterize the melanoma tumor immune microenvironment in mice from single cell to flow cytometry analysis.

Here, we present a protocol to study and describe immune cells that surround or infiltrate tumor cells or get through the body of a melanoma syngeneic mice model. We describe steps for creating and establishing the syngeneic mouse model, euthanasia, and tumor or organ harvest. We then detail procedures to rapidly achieve a single-cell suspension from different tissue samples to further quantify and analyze the phenotype of the immune cell population (lymphocytes T and B, tumor-associated macrophages, and myeloid-derived suppressor cells) by flow cytometry.

ARID1A-deficient cells require HDAC6 for progression of endometrial carcinoma.

AT-rich interactive domain-containing protein 1A (ARID1A) loss-of-function mutation accompanied by a loss of ARID1A protein expression is frequently observed in endometrial carcinomas. However, the molecular mechanisms linking these genetic changes to the altered pathways regulating tumour initiation, maintenance and/or progression remain poorly understood. Thus, the main aim of this study was to analyse the role of ARID1A loss of function in endometrial tumorigenesis. Here, using different endometrial in vitro and in vivo models, such as tumoral cell lines, 3D primary cultures and metastatic or genetically modified mouse models, we show that altered expression of ARID1A is not enough to initiate endometrial tumorigenesis. However, in an established endometrial cancer context, ARID1A loss of function accelerates tumoral progression and metastasis through the disruption of the G2/M cell cycle checkpoint and ATM/ATR-mediated DNA damage checkpoints, increases epithelial cell proliferation rates and induces epithelial mesenchymal transition through the activation of histone deacetylase 6 (HDAC6). Next, we demonstrated that the inhibition of HDAC6 function, using the HDAC6-specific inhibitor ACY1215 or by transfection with HDAC6 short hairpin RNA (shRNA), can reverse the migratory and invasive phenotype of ARID1A-knockdown cells. Further, we also show that inhibition of HDAC6 activity causes an apoptotic vulnerability to etoposide treatments in ARID1A-deficient cells. In summary, the findings exposed in this work indicate that the inhibition of HDAC6 activity is a potential therapeutic strategy for patients suffering from ARID1A-mutant endometrial cancer diagnosed in advanced stages.

BRAFV600E Mutant Allele Frequency (MAF) Influences Melanoma Clinicopathologic Characteristics.

BACKGROUND: Cutaneous melanoma shows high variability regarding clinicopathological presentation, evolution and prognosis. METHODS: Next generation sequencing was performed to analyze hotspot mutations in different areas of primary melanomas (MMp) and their paired metastases. Clinicopathological features were evaluated depending on the degree of variation of the BRAFV600E mutant allele frequency (MAF) in MMp. RESULTS: In our cohort of 14 superficial spreading, 10 nodular melanomas and 52 metastases, 17/24 (71%) melanomas had a BRAFV600E mutation and 5/24 (21%) had a NRASQ61 mutation. We observed a high variation of BRAFV600E MAF (H-BRAFV600E) in 7/17 (41%) MMp. The H-BRAFV600E MMp were all located on the trunk, had lower Breslow and mitotic indexes and predominantly, a first nodal metastasis. Regions with spindled tumor cells (Spin) and high lymphocytic infiltrate (HInf) were more frequent in the H-BRAFV600E patients (4/7; 57%), whereas regions with epithelial tumor cells (Epit) and low lymphocytic infiltrate (LInf) were predominant (6/10; 60%) and exclusive in the low BRAFV600E MAF variation tumors (L-BRAFV600E). The H-BRAFV600E/Spin/HInf MMp patients had better prognostic features and nodal first metastasis. CONCLUSIONS: The H-BRAFV600E MMp were located on the trunk, had better prognostic characteristics, such as lower Breslow and mitotic indexes as well as high lymphocytic infiltrate.

T-Type Calcium Channels: A Potential Novel Target in Melanoma.

T-type calcium channels (TTCCs) are overexpressed in several cancers. In this review, we summarize the recent advances and new insights into TTCC biology, tumor progression, and prognosis biomarker and therapeutic potential in the melanoma field. We describe a novel correlation between the Cav3.1 isoform and the increased basal autophagy in BRAFV600E-mutant melanomas and after acquired resistance to BRAF inhibitors. Indeed, TTCC blockers reduce melanoma cell viability and migration/invasion in vitro and tumor growth in mice xenografts in both BRAF-inhibitor-sensitive and -resistant scenarios. These studies open a new, promising therapeutic approach for disseminated melanoma and improved treatment in BRAFi relapsed melanomas, but further validation and clinical trials are needed for it to become a real therapeutic option.

Retrograde intrarenal surgery (RIRS) in the treatment of calyceal diverticulum with lithiasis.

OBJECTIVES: To evaluate the result of retrograde intrarenal surgery (RIRS) assisted by flexible ureterorenoscopy (FURS) and Holmium laser in the treatment of lithiasis within calyceal diverticula as a minimally invasive therapeutic option. METHODS: We retrospectively evaluated 11 cases of symptomatic lithiasis within calyceal diverticula treated between January 2010 and December 2011. We defined treatment success as absence of residual stones and absence/disappearance of symptomatology over the course of follow-up. We describe the RIRS technique and maneuvers for locating the diverticulum, opening the neck, and fragmenting intradiverticular lithiasis. RESULTS: The most frequently experienced symptom was flank pain (72.7%). The size of the lithiasis treated ranged from 7-20 mm. The overall success rate of RIRS was approximately 73% (absence of lithiasis and disappearance of symptoms) with an average follow-up of 13.3 months. Three cases were not solved by RIRS (2 due to unsuccessful location of the neck, 1 due to persistence of lithiasis and symptoms) . Cases of unsuccessful location were treated with laparoscopic surgery. CONCLUSION: RIRS assisted by FURS and Holmium laser is an effective and minimally invasive procedure for the treatment of lithiasis in the interior of the calyceal diverticulum. This treatment's efficacy improves upon the results from ESWL (extracorporeal shock wave lithotripsy( and equals that of the percutaneous method, exhibiting a lower rate of complications.

Cirugía retrograda intrarenal en el tratamiento del divertículo calicial con litiasis / Retrograde intrarenal surgery (RIRS) in the treatment of calyceal diverticulum with lithiasis

OBJETIVO: Evaluar el resultado de la cirugía retrograda intrarenal (CRIR) con uso de ureterorrenoscopio flexible (URSF) y láser Holmium en el tratamiento de la litiasis en el interior del divertículo calicial como una opción terapéutica mínimamente invasiva. MÉTODOS: Evaluamos retrospectivamente 11 casos de litiasis sintomáticas en el interior de divertículo calicial tratadas entre enero de 2010 y Diciembre de 2011. Definimos éxito del tratamiento: ausencia de restos litiásicos residuales y ausencia/desaparición de la sintomatología a lo largo del seguimiento. Describimos la técnica de CRIR y maniobras para la localización del divertículo, apertura del cuello y fragmentación de litiasis intradiverticular. RESULTADOS: El síntoma de presentación más frecuente fue dolor en el flanco (72,/%). El tamaño de litiasis tratada osciló entre 7-20 mm, El éxito global de CRIR fue de un 73% (ausencia litiasis y desaparición síntomas) con media de seguimiento de 13,3 meses. En 3 casosla CRIR no fue resolutiva (2 por no localización del cuello, 1 por persistencia de litiasis y sintomatología). Los casos de no localización se trataron con cirugía laparoscópica. CONCLUSIÓN: La CRIR con uso de URSF y láser holmium es un procedimiento efectivo y mínimamente invasivo en el manejo de la litiasis en el interior del divertículo calicial

OBJECTIVES: To evaluate the result of retrograde intrarenal surgery (RIRS) assisted by flexible ureterorenoscopy (FURS) and Holmium laser in the treatment of lithiasis within calyceal diverticula as a minimally invasive therapeutic option. METHODS: We retrospectively evaluated 11 cases of symptomatic lithiasis within calyceal diverticula treated between January 2010 and December 2011. We defined treatment success as absence of residual stones and absence/disappearance of symptomatology over the course of follow-up. We describe the RIRS technique and maneuvers for locating the diverticulum, opening the neck, and fragmenting intradiverticular lithiasis. RESULTS: The most frequently experienced symptom was flank pain (72.7 %). The size of the lithiasis treated ranged from 7-20 mm. The overall success rate of RIRS was approximately 73% (absence of lithiasis and disappearance of symptoms) with an average follow-up of 13.3 months. Three cases were not solved by RIRS (2 due to unsuccessful location of the neck, 1 due to persistence of lithiasis and symptoms). Cases of unsuccessful location were treated with laparoscopic surgery. CONCLUSION: RIRS assisted by FURS and Holmium laser is an effective and minimally invasive procedure for the treatment of lithiasis in the interior of the calyceal diverticulum. This treatment’s efficacy improves upon the results from ESWL [extracorporeal shock wave lithotripsy] and equals that of the percutaneous method, exhibiting a lower rate of complications