CONTEXT: Pheochromocytomas and paragangliomas are notable for a high frequency of inherited cases, many of which present as apparently sporadic tumors. OBJECTIVE: The objective of this study was to establish a comprehensive next generation sequencing (NGS)-based strategy for the diagnosis of patients with pheochromocytoma and paraganglioma by testing simultaneously for mutations in MAX, RET, SDHA, SDHB, SDHC, SDHD, SDHAF2, TMEM127, and VHL. DESIGN: After the methodology for the assay was designed and established, it was validated on DNA samples with known genotype and then patients were studied prospectively. SETTING: The study was performed in a diagnostic genetics laboratory. PATIENTS: DNA samples from 205 individuals affected with adrenal or extraadrenal pheochromocytoma/head and neck paraganglioma (PPGL/HNPGL) were analyzed. A proof-of-principle study was performed using 85 samples known to contain a variant in 1 or more of the genes to be tested, followed by prospective analysis of an additional 120 samples. MAIN OUTCOME MEASURES: We assessed the ability to use an NGS-based method to perform comprehensive analysis of genes implicated in inherited PPGL/HNPGL. RESULTS: The proof-of-principle study showed that the NGS assay and analysis gave a sensitivity of 98.7%. A pathogenic mutation was identified in 16.6% of the prospective analysis cohort of 120 patients. CONCLUSIONS: A comprehensive NGS-based strategy for the analysis of genes associated with predisposition to PPGL and HNPGL was established, validated, and introduced into diagnostic service. The new assay provides simultaneous analysis of 9 genes and allows more rapid and cost-effective mutation detection than the previously used conventional Sanger sequencing-based methodology.
CONTEXT: Pheochromocytomas and paragangliomas are notable for a high frequency of inherited cases, many of which present as apparently sporadic tumors. OBJECTIVE: The objective of this study was to establish a comprehensive next generation sequencing (NGS)-based strategy for the diagnosis of patients with pheochromocytoma and paraganglioma by testing simultaneously for mutations in MAX, RET, SDHA, SDHB, SDHC, SDHD, SDHAF2, TMEM127, and VHL. DESIGN: After the methodology for the assay was designed and established, it was validated on DNA samples with known genotype and then patients were studied prospectively. SETTING: The study was performed in a diagnostic genetics laboratory. PATIENTS: DNA samples from 205 individuals affected with adrenal or extraadrenal pheochromocytoma/head and neck paraganglioma (PPGL/HNPGL) were analyzed. A proof-of-principle study was performed using 85 samples known to contain a variant in 1 or more of the genes to be tested, followed by prospective analysis of an additional 120 samples. MAIN OUTCOME MEASURES: We assessed the ability to use an NGS-based method to perform comprehensive analysis of genes implicated in inherited PPGL/HNPGL. RESULTS: The proof-of-principle study showed that the NGS assay and analysis gave a sensitivity of 98.7%. A pathogenic mutation was identified in 16.6% of the prospective analysis cohort of 120 patients. CONCLUSIONS: A comprehensive NGS-based strategy for the analysis of genes associated with predisposition to PPGL and HNPGL was established, validated, and introduced into diagnostic service. The new assay provides simultaneous analysis of 9 genes and allows more rapid and cost-effective mutation detection than the previously used conventional Sanger sequencing-based methodology.
Authors: Iñaki Comino-Méndez; Luis J Leandro-García; Guillermo Montoya; Lucía Inglada-Pérez; Aguirre A de Cubas; María Currás-Freixes; Carolyn Tysoe; Louise Izatt; Rocío Letón; Álvaro Gómez-Graña; Veronika Mancikova; María Apellániz-Ruiz; Massimo Mannelli; Francesca Schiavi; Judith Favier; Anne-Paule Gimenez-Roqueplo; Henri J L M Timmers; Giovanna Roncador; Juan F Garcia; Cristina Rodríguez-Antona; Mercedes Robledo; Alberto Cascón Journal: J Mol Med (Berl) Date: 2015-06-14 Impact factor: 4.599
Authors: David Taïeb; Alexandre Kaliski; Carsten C Boedeker; Victoria Martucci; Tito Fojo; John R Adler; Karel Pacak Journal: Endocr Rev Date: 2014-07-17 Impact factor: 19.871
Authors: Susan Richter; Mirko Peitzsch; Elena Rapizzi; Jacques W Lenders; Nan Qin; Aguirre A de Cubas; Francesca Schiavi; Jyotsna U Rao; Felix Beuschlein; Marcus Quinkler; Henri J Timmers; Giuseppe Opocher; Massimo Mannelli; Karel Pacak; Mercedes Robledo; Graeme Eisenhofer Journal: J Clin Endocrinol Metab Date: 2014-07-11 Impact factor: 5.958