title: DACH1, a zona glomerulosa selective gene in the human adrenal, activates transforming growth factor-β signaling and suppresses aldosterone secretion.
abstract: Common somatic mutations in CACNAID and ATP1A1 may define a subgroup of smaller, zona glomerulosa (ZG)-like aldosterone-producing adenomas. We have therefore sought signature ZG genes, which may provide insight into the frequency and pathogenesis of ZG-like aldosterone-producing adenomas. Twenty-one pairs of zona fasciculata and ZG and 14 paired aldosterone-producing adenomas from 14 patients with Conn's syndrome and 7 patients with pheochromocytoma were assayed by the Affymetrix Human Genome U133 Plus 2.0 Array. Validation by quantitative real-time polymerase chain reaction was performed on genes >10-fold upregulated in ZG (compared with zona fasciculata) and >10-fold upregulated in aldosterone-producing adenomas (compared with ZG). DACH1, a gene associated with tumor progression, was further analyzed. The role of DACH1 on steroidogenesis, transforming growth factor-β, and Wnt signaling activity was assessed in the human adrenocortical cell line, H295R. Immunohistochemistry confirmed selective expression of DACH1 in human ZG. Silencing of DACH1 in H295R cells increased CYP11B2 mRNA levels and aldosterone production, whereas overexpression of DACH1 decreased aldosterone production. Overexpression of DACH1 in H295R cells activated the transforming growth factor-β and canonical Wnt signaling pathways but inhibited the noncanonical Wnt signaling pathway. Stimulation of primary human adrenal cells with angiotensin II decreased DACH1 mRNA expression. Interestingly, there was little overlap between our top ZG genes and those in rodent ZG. In conclusion, (1) the transcriptome profile of human ZG differs from rodent ZG, (2) DACH1 inhibits aldosterone secretion in human adrenals, and (3) transforming growth factor-β signaling pathway is activated in DACH1 overexpressed cells and may mediate inhibition of aldosterone secretion in human adrenals.
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Junhua
authors.initials: J
authors.lastname: Zhou
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Lalarukh Haris
authors.initials: LH
authors.lastname: Shaikh
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Sudeshna G
authors.initials: SG
authors.lastname: Neogi
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Ian
authors.initials: I
authors.lastname: McFarlane
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Wanfeng
authors.initials: W
authors.lastname: Zhao
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Nichola
authors.initials: N
authors.lastname: Figg
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Brighton
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Carmela
authors.initials: C
authors.lastname: Maniero
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Ada E D
authors.initials: AE
authors.lastname: Teo
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.).
authors.forename: Elena A B
authors.initials: EA
authors.lastname: Azizan
authors.affiliation: From the Clinical Pharmacology Unit, Department of Medicine (J.Z., L.H.S., C.A.B., C.M., A.E.D.T, E.A.B.A., M.J.B.), Cardiovascular Division, Department of Medicine (N.F.), University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom; Department of Clinical Biochemistry, GenomicsCoreLab, Cambridge NIHR BRC, Department of Clinical Biochemistry (S.G.N., I.M.), and Human Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust (W.Z.), Addenbrooke's Hospital, Cambridge, United Kingdom; and Department of Medicine, Faculty of Medicine, The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia (E.A.B.A.). m.j.brown@cai.cam.ac.uk.
authors.forename: Morris J
authors.initials: MJ
authors.lastname: Brown
mesh_terms: D018268:Adrenocortical Carcinoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000450:Aldosterone / Q000378:metabolism*; D018450:Disease Progression; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D012334:RNA, Neoplasm / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics*; D014407:Tumor Cells, Cultured; D015384:Zona Glomerulosa / Q000378:metabolism* / Q000473:pathology
chemical_list: C486747:DACH1 protein, human; D005136:Eye Proteins; D012334:RNA, Neoplasm; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D000450:Aldosterone
keywords: DACH1 protein, human; aldosterone; hyperaldosteronism; transforming growth factor beta; zona glomerulosa