A Comprehensive Next Generation Sequencing-Based Genetic Testing Strategy to Improve Diagnosis of Inherited Pheochromocytoma and Paraganglioma

J Clin Endocrinol Metab. 2013 Jul;98(7):E1248-56. doi: 10.1210/jc.2013-1319. Epub 2013 May 10.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adrenal Gland Neoplasms / diagnosis*
  • Adrenal Gland Neoplasms / economics
  • Adrenal Gland Neoplasms / genetics
  • Adrenal Gland Neoplasms / metabolism
  • Cohort Studies
  • Cost Savings
  • Costs and Cost Analysis
  • DNA Mutational Analysis / economics
  • Genetic Predisposition to Disease
  • Genetic Testing / economics
  • Genetic Testing / methods
  • Germ-Line Mutation*
  • Head and Neck Neoplasms / diagnosis*
  • Head and Neck Neoplasms / economics
  • Head and Neck Neoplasms / genetics
  • Head and Neck Neoplasms / metabolism
  • Health Care Costs
  • Humans
  • Paraganglioma / diagnosis*
  • Paraganglioma / economics
  • Paraganglioma / genetics
  • Paraganglioma / metabolism
  • Pheochromocytoma / diagnosis*
  • Pheochromocytoma / economics
  • Pheochromocytoma / genetics
  • Pheochromocytoma / metabolism
  • Prospective Studies
  • Protein Subunits / chemistry
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • Proto-Oncogene Proteins c-ret / chemistry
  • Proto-Oncogene Proteins c-ret / genetics
  • Proto-Oncogene Proteins c-ret / metabolism
  • Sensitivity and Specificity
  • Succinate Dehydrogenase / chemistry
  • Succinate Dehydrogenase / genetics
  • Succinate Dehydrogenase / metabolism
  • United Kingdom
  • Von Hippel-Lindau Tumor Suppressor Protein / chemistry
  • Von Hippel-Lindau Tumor Suppressor Protein / genetics
  • Von Hippel-Lindau Tumor Suppressor Protein / metabolism

Substances

  • Protein Subunits
  • Succinate Dehydrogenase
  • Von Hippel-Lindau Tumor Suppressor Protein
  • Proto-Oncogene Proteins c-ret
  • RET protein, human
  • VHL protein, human