RESUMEN
Molecular diagnostics are increasingly performed routinely in the diagnosis and management of patients with melanoma due to the development of novel therapies that target specific genetic mutations. The development of next-generation sequencing (NGS) technologies has enabled to sequence multiple cancer-driving genes in a single assay, with improved sensitivity in mutation detection. The main objective of this study was the design and implementation of a melanoma-specific sequencing panel, and the identification of the spectrum of somatic mutations in a series of primary melanoma samples. A custom panel was designed to cover the coding regions of 35 melanoma-related genes. Panel average coverage was 2,575.5 reads per amplicon, with 92,8% of targeted bases covered ≥500×. Deep coverage enabled sensitive discovery of mutations in as low as 0.5% mutant allele frequency. Eighty-five percent (85/100) of the melanomas had at least one somatic mutation. The most prevalent mutated genes were BRAF (50%;50/199), NRAS (15%;15/100), PREX2 (14%;14/100), GRIN2A (13%;13/100), and ERBB4 (12%;12/100). Turn-around-time and costs for NGS-based analysis was reduced in comparison to conventional molecular approaches. The results of this study demonstrate the cost-effectiveness and feasibility of a custom-designed targeted NGS panel, and suggest the implementation of targeted NGS into daily routine practice.
Asunto(s)
Melanoma/diagnóstico , Melanoma/genética , Medicina de Precisión , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor , Línea Celular Tumoral , Análisis Costo-Beneficio , Femenino , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Pruebas Genéticas/métodos , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Melanoma/terapia , Persona de Mediana Edad , Mutación , Oportunidad Relativa , Sensibilidad y EspecificidadAsunto(s)
Adenocarcinoma/secundario , Neoplasias del Pene/secundario , Neoplasias de la Próstata/patología , Neoplasias Cutáneas/secundario , Adenocarcinoma/terapia , Diagnóstico Diferencial , Humanos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Pelvis , Neoplasias del Pene/diagnóstico , Neoplasias del Pene/tratamiento farmacológico , Neoplasias de la Próstata/terapia , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/tratamiento farmacológicoRESUMEN
BACKGROUND AND OBJECTIVE: Several clinical and histological prognostic factors have been identified in localized melanoma. However, further studies with better defined and more reproducible histological parameters are needed. Our aim was to identify the prognostic factors for survival in cutaneous melanoma in the Spanish population. PATIENTS AND METHOD: Six hundred and thirty nine patients with localized melanoma, stages I and II of the last version of the American Joint Committee on Cancer staging system for cutaneous melanoma, with 2 years of follow-up or documented relapse, were selected from the database of the Department of Dermatology. Univariate and multivariate Cox regression analyses were performed for overall and disease free survival. RESULTS: Tumor thickness appeared as the most important prognostic factor for both overall and disease free survival in the multivariate analysis. Inflammatory infiltrate and sex were only significant for overall survival, and location, age and ulceration were significant for disease free survival. Other variables, such as histological type, mitotic rate or level of invasion, lost their prognostic significance in the multivariate analysis. CONCLUSIONS: Tumor thickness is the most important prognostic factor to predict survival in localized melanoma. Other factors such as sex, inflammatory infiltrate, location, age or ulceration, have also an important role in prognosis.
