Rare germline mutations identified by targeted next-generation sequencing of susceptibility genes in pheochromocytoma and paraganglioma.

CONTEXT Pheochromocytomas and paragangliomas have a highly diverse genetic background, with a third of the cases carrying a germline mutation in 1 of 14 identified genes. OBJECTIVE This study aimed to evaluate next-generation sequencing for more efficient genetic testing of pheochromocytoma and paraganglioma and to establish germline and somatic mutation frequencies for all known susceptibility genes. DESIGN A targeted next-generation sequencing approach on an Illumina MiSeq instrument was used for a mutation analysis in 86 unselected pheochromocytoma and paraganglioma tumor samples. The study included the genes EGLN1, EPAS1, KIF1Bβ, MAX, MEN1, NF1, RET, SDHA, SDHB, SDHC, SDHD, SDHAF2, TMEM127, and VHL. RESULTS were verified in tumor and constitutional DNA with Sanger sequencing. RESULTS In all cases with clinical syndromes or known germline mutations, a mutation was detected in the expected gene. Among 68 nonfamilial tumors, 32 mutations were identified in 28 of the samples (41%), including germline mutations in EGLN1, KIF1Bβ, SDHA, SDHB, and TMEM127 and somatic mutations in EPAS1, KIF1Bβ, MAX, NF1, RET, and VHL, including one double monoallelic EPAS1 mutation. CONCLUSIONS Targeted next-generation sequencing proved to be fast and cost effective for the genetic analysis of pheochromocytoma and paraganglioma. More than half of the tumors harbored mutations in the investigated genes. Notably, 7% of the apparently sporadic cases carried germline mutations, highlighting the importance of comprehensive genetic testing. KIF1Bβ, which previously has not been investigated in a large cohort, appears to be an equally important tumor suppressor as MAX and TMEM127 and could be considered for genetic testing of these patients.

[1]  P. Rustin,et al.  Germline mutations in FH confer predisposition to malignant pheochromocytomas and paragangliomas. , 2014, Human molecular genetics.

[2]  R. Sandberg,et al.  RNA helicase A is a downstream mediator of KIF1Bβ tumor-suppressor function in neuroblastoma. , 2014, Cancer discovery.

[3]  P. Dahia.  Pheochromocytoma and paraganglioma pathogenesis: learning from genetic heterogeneity , 2014, Nature Reviews Cancer.

[4]  M. Marshall,et al.  Whole exome sequencing is an efficient and sensitive method for detection of germline mutations in patients with phaeochromcytomas and paragangliomas , 2014, Clinical endocrinology.

[5]  A. Barlier,et al.  A MEN1 syndrome with a paraganglioma , 2013, European Journal of Human Genetics.

[6]  P. Söderkvist,et al.  The NF1 gene: a frequent mutational target in sporadic pheochromocytomas and beyond. , 2013, Endocrine-related cancer.

[7]  Laurence Amar,et al.  SDH mutations establish a hypermethylator phenotype in paraganglioma. , 2013, Cancer cell.

[8]  P. Söderkvist,et al.  Germline SDHA mutation detected by next-generation sequencing in a young index patient with large paraganglioma. , 2013, The Journal of clinical endocrinology and metabolism.

[9]  S. Srikantan,et al.  In vivo and in vitro oncogenic effects of HIF2A mutations in pheochromocytomas and paragangliomas. , 2013, Endocrine-related cancer.

[10]  P. Hellman,et al.  Next-generation sequencing in the clinical genetic screening of patients with pheochromocytoma and paraganglioma , 2013, Endocrine connections.

[11]  M. Korbonits,et al.  A comprehensive next generation sequencing-based genetic testing strategy to improve diagnosis of inherited pheochromocytoma and paraganglioma. , 2013, The Journal of clinical endocrinology and metabolism.

[12]  P. Hellman,et al.  Somatic mutations in H-RAS in sporadic pheochromocytoma and paraganglioma identified by exome sequencing. , 2013, The Journal of clinical endocrinology and metabolism.

[13]  K. Pacak,et al.  First report of bilateral pheochromocytoma in the clinical spectrum of HIF2A-related polycythemia-paraganglioma syndrome. , 2013, The Journal of clinical endocrinology and metabolism.

[14]  D. Fraker,et al.  Inherited Mutations in Pheochromocytoma and Paraganglioma: Why All Patients Should Be Offered Genetic Testing , 2013, Annals of Surgical Oncology.

[15]  G. Pita,et al.  Tumoral EPAS1 (HIF2A) mutations explain sporadic pheochromocytoma and paraganglioma in the absence of erythrocytosis. , 2013, Human molecular genetics.

[16]  C. Larsson,et al.  Acquired hypermethylation of the P16INK4A promoter in abdominal paraganglioma: relation to adverse tumor phenotype and predisposing mutation , 2012, Endocrine-related cancer.

[17]  E. Letouzé,et al.  Somatic NF1 inactivation is a frequent event in sporadic pheochromocytoma. , 2012, Human molecular genetics.

[18]  C. Larsson,et al.  Integrative genomics reveals frequent somatic NF1 mutations in sporadic pheochromocytomas. , 2012, Human molecular genetics.

[19]  J. Prchal,et al.  A novel EPAS1/HIF2A germline mutation in a congenital polycythemia with paraganglioma , 2012, Journal of Molecular Medicine.

[20]  Electron Kebebew,et al.  Somatic HIF2A gain-of-function mutations in paraganglioma with polycythemia. , 2012, The New England journal of medicine.

[21]  M. Urioste,et al.  MAX Mutations Cause Hereditary and Sporadic Pheochromocytoma and Paraganglioma , 2012, Clinical Cancer Research.

[22]  P. Söderkvist,et al.  Genetics and clinical characteristics of hereditary pheochromocytomas and paragangliomas. , 2011, Endocrine-related cancer.

[23]  Heikki Joensuu,et al.  Comparison of solution-based exome capture methods for next generation sequencing , 2011, Genome Biology.

[24]  A. Vénisse,et al.  Integrative genomic analysis reveals somatic mutations in pheochromocytoma and paraganglioma. , 2011, Human molecular genetics.

[25]  W. Dinjens,et al.  SDHA immunohistochemistry detects germline SDHA gene mutations in apparently sporadic paragangliomas and pheochromocytomas. , 2011, The Journal of clinical endocrinology and metabolism.

[26]  J. Benítez,et al.  Exome sequencing identifies MAX mutations as a cause of hereditary pheochromocytoma , 2011, Nature Genetics.

[27]  Helga Thorvaldsdóttir,et al.  Integrative Genomics Viewer , 2011, Nature Biotechnology.

[28]  C. Ricketts,et al.  Mutation analysis of HIF prolyl hydroxylases (PHD/EGLN) in individuals with features of phaeochromocytoma and renal cell carcinoma susceptibility , 2010, Endocrine-related cancer.

[29]  S. Gruber,et al.  Spectrum and prevalence of FP/TMEM127 gene mutations in pheochromocytomas and paragangliomas. , 2010, JAMA.

[30]  Jana Marie Schwarz,et al.  MutationTaster evaluates disease-causing potential of sequence alterations , 2010, Nature Methods.

[31]  P. Bork,et al.  A method and server for predicting damaging missense mutations , 2010, Nature Methods.

[32]  Patricia L. M. Dahia,et al.  Germline mutations in TMEM127 confer susceptibility to pheochromocytoma , 2010, Nature Genetics.

[33]  F. Galateau-Sallé,et al.  PHD2 mutation and congenital erythrocytosis with paraganglioma. , 2008, The New England journal of medicine.

[34]  A. Ligon,et al.  A germline mutation of the KIF1Bβ gene on 1p36 in a family with neural and nonneural tumors , 2008, Human Genetics.

[35]  C. Larsson,et al.  Global and Regional CpG Methylation in Pheochromocytomas and Abdominal Paragangliomas: Association to Malignant Behavior , 2008, Clinical Cancer Research.

[36]  M. Meyerson,et al.  The kinesin KIF1Bbeta acts downstream from EglN3 to induce apoptosis and is a potential 1p36 tumor suppressor. , 2008, Genes & development.

[37]  Patrick D. Sutphin,et al.  Coordinate Regulation of the Oxygen-Dependent Degradation Domains of Hypoxia-Inducible Factor 1α , 2005, Molecular and Cellular Biology.

[38]  P. Rustin,et al.  Mutations in the SDHB gene are associated with extra-adrenal and/or malignant phaeochromocytomas. , 2003, Cancer research.

[39]  W. Simonds,et al.  Multiple endocrine neoplasia type 1: new clinical and basic findings , 2001, Trends in Endocrinology & Metabolism.

[40]  C. Larsson,et al.  Comparative genomic hybridization reveals frequent losses of chromosomes 1p and 3q in pheochromocytomas and abdominal paragangliomas, suggesting a common genetic etiology. , 2000, The American journal of pathology.

[41]  Masahide Takahashi,et al.  Somatic Mutations in RET Exons 12 and 15 in Sporadic Medullary Thyroid Carcinomas: Different Spectrum of Mutations in Sporadic Type from Hereditary Type , 1999, Japanese journal of cancer research : Gann.