title: Towards Personalized Medicine in Melanoma: Implementation of a Clinical Next-Generation Sequencing Panel.
abstract: 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.
authors.affiliation: Department of Dermatology, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Blanca
authors.initials: B
authors.lastname: de Unamuno Bustos
authors.affiliation: Molecular Biology Laboratory, Service of Clinical Analysis, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Murria Estal
authors.affiliation: Molecular Biology Laboratory, Service of Clinical Analysis, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Gema
authors.initials: G
authors.lastname: Pérez Simó
authors.affiliation: Molecular Biology Laboratory, Service of Clinical Analysis, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: de Juan Jimenez
authors.affiliation: Department of Dermatology, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Begoña
authors.initials: B
authors.lastname: Escutia Muñoz
authors.affiliation: Department of Dermatology, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Rodríguez Serna
authors.affiliation: Department of Dermatology, Hospital General Universitario de Valencia, Valencia, Spain.
authors.forename: Victor
authors.initials: V
authors.lastname: Alegre de Miquel
authors.affiliation: Department of Pathology, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Margarita
authors.initials: M
authors.lastname: Llavador Ros
authors.affiliation: Department of Dermatology, La Plana Hospital, Villarreal, Castellón, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Ballester Sánchez
authors.affiliation: Department of Dermatology, Department of Dermatology, Instituto Valenciano de Oncología, Valencia, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Nagore Enguídanos
authors.affiliation: Molecular Biology Laboratory, Service of Clinical Analysis, Hospital Universitari i Politecnic La Fe, Valencia, Spain. palanca_sar@gva.es.
authors.forename: Sarai
authors.initials: S
authors.lastname: Palanca Suela
authors.affiliation: Department of Dermatology, Hospital Universitari i Politecnic La Fe, Valencia, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Botella Estrada
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor; D045744:Cell Line, Tumor; D003362:Cost-Benefit Analysis; D005260:Female; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008545:Melanoma / Q000175:diagnosis* / Q000235:genetics* / Q000628:therapy; D008875:Middle Aged; D009154:Mutation; D016017:Odds Ratio; D057285:Precision Medicine; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor