title: Double adrenocortical adenomas harboring independent KCNJ5 and PRKACA somatic mutations.
abstract: OBJECTIVE
  Co-secretion of cortisol and aldosterone can be observed in adrenal adenomas. The aim of this study was to investigate the molecular characteristics of a co-existing aldosterone- and a cortisol-producing adenoma (CPA) in the same patient.
  
  
  METHODS
  Two different adenomas within the same adrenal gland from a 49-year-old female patient with primary aldosteronism (PA) and Cushing's syndrome (CS) were studied. Multiple formalin-fixed paraffin-embedded tumor blocks were used for the analysis. Immunohistochemistry (IHC) was performed using a specific antibody against aldosterone synthase (CYP11B2). DNA and RNA were isolated separately from CYP11B2-positive and -negative tumor regions based on CYP11B2 IHC results.
  
  
  RESULTS
  CYP11B2 IHC clearly demonstrated that three pieces from one adenoma were positive for CYP11B2 and the remaining three from the other adenoma were negative for CYP11B2. In quantitative real-time RT-PCR, CYP11B2 mRNA was upregulated in CYP11B2-positive tumor specimens (219-fold vs CYP11B2-negative tumor specimens). Targeted next-generation sequencing (NGS) detected novel KCNJ5 gene mutations (p.T148I/T149S, present in the same reads) and a PRKACA gene hotspot mutation (p.L206R) in the CYP11B2-positive and -negative tumors, respectively. Sanger sequencing of DNA from each tumor specimen (CYP11B2-positive tumor, n=3; CYP11B2-negative tumor, n=3) showed concordant results with targeted NGS.
  
  
  CONCLUSIONS
  Our findings illustrate the co-existence of two different adrenocortical adenomas causing the concurrent diagnosis of PA and CS in the same patient. Molecular analysis was able to demonstrate that the two diseases resulted from independent somatic mutations seen in double adrenocortical adenomas.
authors.affiliation: Departments of Molecular and Integrative Physiology & Internal Medicine.
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Nanba
authors.affiliation: Department of Pathology.
authors.forename: Kei
authors.initials: K
authors.lastname: Omata
authors.affiliation: Department of Pathology Comprehensive Cancer Center Michigan Center for Translational Pathology Department of Urology.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Tomlins
authors.affiliation: Department of Pathology Comprehensive Cancer Center Division of Metabolism, Endocrinology, and Diabetes.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Giordano
authors.affiliation: Departments of Molecular and Integrative Physiology & Internal Medicine Division of Metabolism, Endocrinology, and Diabetes Endocrine Oncology ProgramCenter for Organogenesis, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Gary D
authors.initials: GD
authors.lastname: Hammer
authors.affiliation: Departments of Molecular and Integrative Physiology & Internal Medicine.
authors.forename: William E
authors.initials: WE
authors.lastname: Rainey
authors.affiliation: Division of Metabolism, Endocrinology, and Diabetes telse@umich.edu.
authors.forename: Tobias
authors.initials: T
authors.lastname: Else
mesh_terms: D000306:Adrenal Cortex Neoplasms / Q000097:blood / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D018246:Adrenocortical Adenoma / Q000097:blood / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D000450:Aldosterone / Q000097:blood; D003480:Cushing Syndrome / Q000097:blood / Q000235:genetics / Q000473:pathology / Q000601:surgery; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / Q000235:genetics*; D005260:Female; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels / Q000235:genetics*; D006801:Humans; D006854:Hydrocortisone / Q000097:blood; D006929:Hyperaldosteronism / Q000097:blood / Q000235:genetics / Q000473:pathology / Q000601:surgery; D008875:Middle Aged; D009154:Mutation; D009378:Neoplasms, Multiple Primary / Q000097:blood / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D016896:Treatment Outcome
chemical_list: D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; C484274:KCNJ5 protein, human; D000450:Aldosterone; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits; C517757:PRKACA protein, human; D006854:Hydrocortisone