title: Molecular Heterogeneity in Aldosterone-Producing Adenomas.
abstract: BACKGROUND
  The use of next-generation sequencing has resulted in the identification of recurrent somatic mutations underlying primary aldosteronism (PA). However, significant gaps remain in our understanding of the relationship between tumor aldosterone synthase (CYP11B2) expression and somatic mutation status.
  
  
  OBJECTIVE
  The objective of the study was to investigate tumor CYP11B2 expression and somatic aldosterone-driver gene mutation heterogeneity.
  
  
  METHODS
  Fifty-one adrenals from 51 PA patients were studied. Immunohistochemistry for CYP11B2 was performed. Aldosterone-producing adenomas with intratumor CYP11B2 heterogeneity were analyzed for mutation status using targeted next-generation sequencing. DNA was isolated from CYP11B2-positive, CYP11B2-negative, and adjacent normal areas from formalin-fixed, paraffin-embedded sections.
  
  
  RESULTS
  Of 51 adrenals, seven (14 %) showed distinct heterogeneity in CYP11B2 by immunohistochemistry, including six adenomas with intratumor heterogeneity and one multinodular hyperplastic adrenal with both CYP11B2-positive and -negative nodules. Of the six adrenocortical adenomas with CYP11B2 heterogeneity, three had aldosterone-regulating mutations (CACNA1D p.F747C, KCNJ5 p.L168R, ATP1A1 p.L104R) only in CYP11B2-positive regions, and one had two different mutations localized to two histologically distinct CYP11B2-positive regions (ATP2B3 p.L424_V425del, KCNJ5 p.G151R). Lastly, one adrenal with multiple CYP11B2-expressing nodules showed different mutations in each (CACNA1D p.F747V and ATP1A1 p.L104R), and no mutations were identified in CYP11B2-negative nodule or adjacent normal adrenal.
  
  
  CONCLUSIONS
  Adrenal tumors in patients with PA can demonstrate clear heterogeneity in CYP11B2 expression and somatic mutations in driver genes for aldosterone production. These findings suggest that aldosterone-producing adenoma tumorigenesis can occur within preexisting nodules through the acquisition of somatic mutations that drive aldosterone production.
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Nanba
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Andrew X
authors.initials: AX
authors.lastname: Chen
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Kei
authors.initials: K
authors.lastname: Omata
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Michelle
authors.initials: M
authors.lastname: Vinco
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Giordano
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Tobias
authors.initials: T
authors.lastname: Else
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Gary D
authors.initials: GD
authors.lastname: Hammer
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Tomlins
authors.affiliation: Departments of Molecular and Integrative Physiology and Internal Medicine (K.N., A.X.C., G.D.H., W.E.R.), Pathology (K.O., M.V., T.J.G., S.A.T.), and Urology (S.A.T.), Comprehensive Cancer Center (T.J.G., T.E., S.A.T.), Division of Metabolism, Endocrinology, and Diabetes (T.J.G., T.E., G.D.H.), Endocrine Oncology Program (T.E., G.D.H.), Center for Organogenesis, and Michigan Center for Translational Pathology (S.A.T.), University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: William E
authors.initials: WE
authors.lastname: Rainey
mesh_terms: D000311:Adrenal Glands / Q000473:pathology; D000315:Adrenalectomy; D018246:Adrenocortical Adenoma / Q000737:chemistry* / Q000235:genetics* / Q000473:pathology; D000450:Aldosterone / Q000096:biosynthesis*; D019405:Cytochrome P-450 CYP11B2 / Q000235:genetics*; D004247:DNA / Q000032:analysis / Q000737:chemistry; D015870:Gene Expression; D018740:Genetic Heterogeneity; D006801:Humans; D006929:Hyperaldosteronism / Q000235:genetics* / Q000473:pathology; D007150:Immunohistochemistry; D009154:Mutation; D017422:Sequence Analysis, DNA
chemical_list: D000450:Aldosterone; D004247:DNA; D019405:Cytochrome P-450 CYP11B2