Microarray, qPCR, and KCNJ5 sequencing of aldosterone-producing adenomas reveal differences in genotype and phenotype between zona glomerulosa- and zona fasciculata-like tumors.

CONTEXT Aldosterone-producing adenomas (APA) are heterogeneous. The recent finding of somatic KCNJ5 mutations suggests a genetic explanation. OBJECTIVES The objectives of this study were the following: 1) to compare transcriptional profiles in APA and adjacent adrenal gland (AAG); 2) to test whether gene expression profile clusters with different cell histology; and 3) to measure the frequency of KCNJ5 mutations and determine the genotype-phenotype relationship. DESIGN/SETTING The design of the study included laboratory analyses of 46 unselected APA. PATIENTS The patients in this study had primary hyperaldosteronism with unilateral APA. INTERVENTIONS The objectives of this study were the following: 1) Illumina beadchip analysis of RNA from eight paired APA-AAG; 2) a blinded review of cell histology for 46 APA; 3) laser capture microdissection of zona glomerulosa (ZG) and zona fasciculata (ZF) cells; and 4) sequencing of KCNJ5 in 46 APA. MAIN OUTCOME MEASURES The main outcome measures of this study were the following: 1) a difference in gene expression profile and a correlation with histological markers of ZF; 2) a frequency of KCNJ5 mutations and phenotypic comparisons of wild type with mutant APA. RESULTS The results of the study were the following: 1) a cluster analysis of microarray data separated APA from AAG. APA at opposite ends of the APA cluster had an approximately 800-fold difference in CYP17A1 mRNA expression, whereas histology showed 0% ZF-like cells in one vs. 100% in the other. A heat map ranking APA by CYP17A1 expression correctly predicted several genes (e.g. KCNK1, SLC24A3) to be enriched in laser capture microdissection samples of ZG; 2) known or novel mutations of KCNJ5 were found in 20 of 46 consecutive APA [43% (95% confidence interval [CI] (29, 58)%)]. The APA with KCNJ5 gene mutations were larger compared with tumors harboring the wild type, 1.63 [95% CI (1.37, 1.88)] vs. 1.14 [0.97, 1.30] cm (P = 0.0013), had predominantly ZF-like cells, and their CYP17A1 (log(2)-fold change) was higher than in wild type: -0.96 [95% CI (-0.07, -1.85)] vs. -2.54 [-1.61, -3.46], (P = 0.017). CONCLUSIONS KCNJ5 mutations are common in APA, particularly those arising from ZF. The long-recognized heterogeneity among APA may have a genetic basis.

[1]  Morris J. Brown,et al.  Somatic Mutations Affecting the Selectivity Filter of KCNJ5 Are Frequent in 2 Large Unselected Collections of Adrenal Aldosteronomas , 2012, Hypertension.

[2]  Tom R. Gaunt,et al.  Genetic Variants in Novel Pathways Influence Blood Pressure and Cardiovascular Disease Risk , 2011, Nature.

[3]  G. Rossi A comprehensive review of the clinical aspects of primary aldosteronism , 2011, Nature Reviews Endocrinology.

[4]  H. Sasano,et al.  Gene expression profiles in aldosterone-producing adenomas and adjacent adrenal glands. , 2011, European journal of endocrinology.

[5]  B. Mariniello,et al.  A case of primary aldosteronism in pregnancy: do LH and GNRH receptors have a potential role in regulating aldosterone secretion? , 2011, European journal of endocrinology.

[6]  S. Mane,et al.  K+ Channel Mutations in Adrenal Aldosterone-Producing Adenomas and Hereditary Hypertension , 2011, Science.

[7]  C. Gomez-Sanchez,et al.  Aldosterone-producing adenomas: mining for genes. , 2010, Hypertension.

[8]  W. Rainey,et al.  The role of TASK1 in aldosterone production and its expression in normal adrenal and aldosterone‐producing adenomas , 2009, Clinical endocrinology.

[9]  P. O’Reilly,et al.  Faculty Opinions recommendation of Genome-wide association study identifies eight loci associated with blood pressure. , 2009 .

[10]  G. Altavilla,et al.  Zona fasciculata-like histotype and aldosterone response to upright posture are not related in aldosterone-producing adenomas. , 2009, Experimental and clinical endocrinology & diabetes : official journal, German Society of Endocrinology [and] German Diabetes Association.

[11]  Andrew D. Johnson,et al.  Genome-wide association study of blood pressure and hypertension , 2009, Nature Genetics.

[12]  H. Sakai,et al.  Primary Aldosteronism with Aldosterone-Producing Adenoma Consisting of Pure Zona Glomerulosa-Type Cells in a Pregnant Woman , 2009, Endocrine pathology.

[13]  R. Lifton,et al.  A novel form of human mendelian hypertension featuring nonglucocorticoid-remediable aldosteronism. , 2008, The Journal of clinical endocrinology and metabolism.

[14]  P. Bernante,et al.  Heterogeneity of Aldosterone-Producing Adenomas Revealed by a Whole Transcriptome Analysis , 2007, Hypertension.

[15]  Ping Ye,et al.  G-protein-coupled receptors in aldosterone-producing adenomas: a potential cause of hyperaldosteronism. , 2007, The Journal of endocrinology.

[16]  P. White,et al.  Elevated expression of luteinizing hormone receptor in aldosterone-producing adenomas. , 2006, The Journal of clinical endocrinology and metabolism.

[17]  F. Veglio,et al.  Genetics of primary aldosteronism , 2004, Journal of hypertension.

[18]  D. Calhoun,et al.  Hyperaldosteronism Among Black and White Subjects With Resistant Hypertension , 2002, Hypertension.

[19]  J. Mathis,et al.  Quantitative assessment of CYP11B1 and CYP11B2 expression in aldosterone-producing adenomas. , 2002, European journal of endocrinology.

[20]  D. Farley,et al.  Primary Aldosteronism: Factors Associated with Normalization of Blood Pressure after Surgery , 2001, Annals of Internal Medicine.

[21]  H. Suzuki Pathophysiology and diagnosis of primary aldosteronism. , 2000, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[22]  K. Wu,et al.  Differential expression of type 1 angiotensin II receptor mRNA and aldosterone responsiveness to angiotensin in aldosterone-producing adenoma , 1999, Molecular and Cellular Endocrinology.

[23]  J. Chu,et al.  Zona fasciculata-like cells determine the response of plasma aldosterone to metoclopramide and aldosterone synthase messenger ribonucleic acid level in aldosterone-producing adenoma. , 1995, The Journal of clinical endocrinology and metabolism.

[24]  M. Stowasser,et al.  Primary aldosteronism: hypertension with a genetic basis , 1992, The Lancet.

[25]  A. Ganguly,et al.  Cellular origin of aldosteronomas , 1992, The clinical investigator.

[26]  J. Lalouel,et al.  A chimaeric llβ-hydroxylase/aldosterone synthase gene causes glucocorticoid-remediable aldosteronism and human hypertension , 1992, Nature.

[27]  R. Gordon,et al.  Histological and biochemical distinctiveness of atypical aldosterone‐producing adenomas responsive to upright posture and angiotensin , 1991, Clinical endocrinology.

[28]  H. Sasano,et al.  Immunohistochemical study of cytochrome P-45017α in human adrenocortical disorders , 1989 .

[29]  R. Gordon,et al.  ALDOSTERONE‐PRODUCING ADENOMAS RESPONSIVE TO ANGIOTENSIN POSE PROBLEMS IN DIAGNOSIS , 1987, Clinical and experimental pharmacology & physiology.

[30]  A. Neville,et al.  Histopathology of the human adrenal cortex. , 1985, Clinics in endocrinology and metabolism.

[31]  J. Douglas,et al.  Angiotensin II receptors and in vitro aldosterone responses of aldosterone-producing adenomas, adjacent nontumorous tissue, and normal human adrenal glomerulosa. , 1980, The Journal of clinical endocrinology and metabolism.

[32]  D. Sutherland,et al.  Hypertension, increased aldosterone secretion and low plasma renin activity relieved by dexamethasone. , 1966, Canadian Medical Association journal.

[33]  K. Nakagawa,et al.  Adrenocortical zonation in humans under normal and pathological conditions. , 2010, The Journal of clinical endocrinology and metabolism.

[34]  Ning Guang Expression of aldosterone synthesis related enzyme and associated regulatory factor genes in aldosterone-producing adenoma , 2009 .

[35]  G. Arnaldi,et al.  0021-972X/05/$15.00/0 The Journal of Clinical Endocrinology & Metabolism 90(9):5446–5455 Printed in U.S.A. Copyright © 2005 by The Endocrine Society doi: 10.1210/jc.2005-0836 Expression Profiles for Steroidogenic Enzymes in Adrenocortical Disease , 2005 .

[36]  R. Ardaillou Angiotensin II receptors. , 1999, Journal of the American Society of Nephrology : JASN.