{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T16:59:58Z","timestamp":1791305998370,"version":"4.3.1"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,9,11]],"date-time":"2017-09-11T00:00:00Z","timestamp":1505088000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,9,11]],"date-time":"2017-09-11T00:00:00Z","timestamp":1505088000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Adrenocortical hormone excess, due to primary aldosteronism (PA) or hypercortisolemia, causes hypertension and cardiovascular complications. In PA, hypomethylation of aldosterone synthase (<jats:italic>CYP11B2<\/jats:italic>) is associated with aldosterone overproduction. However, in hypercortisolemia, the role of DNA methylation of 11\u03b2-hydroxylase (<jats:italic>CYP11B1<\/jats:italic>), which catalyzes cortisol biosynthesis and is highly homologous to <jats:italic>CYP11B2<\/jats:italic>, is unclear. The aims of our study were to determine whether the <jats:italic>CYP11B1<\/jats:italic> expression was regulated through DNA methylation in hypercortisolemia with cortisol-producing adenoma (CPA), and to investigate a possible relationship between DNA methylation and somatic mutations identified in CPA. Methylation analysis showed that the <jats:italic>CYP11B1<\/jats:italic> promoter was significantly less methylated in CPA than in adjacent unaffected adrenal tissue and white blood cells. Furthermore, in CPA with somatic mutations in either the catalytic subunit of protein kinase A (<jats:italic>PRKACA<\/jats:italic>) or the guanine nucleotide-binding protein subunit alpha (<jats:italic>GNAS<\/jats:italic>) gene, the <jats:italic>CYP11B1<\/jats:italic> promoter was significantly hypomethylated. In addition, DNA methylation reduced <jats:italic>CYP11B1<\/jats:italic> promoter activity using a reporter assay. Our study results suggest that DNA methylation at the <jats:italic>CYP11B1<\/jats:italic> promoter plays a role in the regulation of <jats:italic>CYP11B1<\/jats:italic> expression and cortisol production in CPA, and that somatic mutations associated with CPA reduce DNA methylation at the <jats:italic>CYP11B1<\/jats:italic> promoter.<\/jats:p>","DOI":"10.1038\/s41598-017-11435-2","type":"journal-article","created":{"date-parts":[[2017,9,5]],"date-time":"2017-09-05T14:50:18Z","timestamp":1504623018000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Cortisol overproduction results from DNA methylation of CYP11B1 in hypercortisolemia"],"prefix":"10.1038","volume":"7","author":[{"given":"Mitsuhiro","family":"Kometani","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takashi","family":"Yoneda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masashi","family":"Demura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Koide","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koshiro","family":"Nishimoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuniaki","family":"Mukai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Celso E.","family":"Gomez-Sanchez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tadayuki","family":"Akagi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takashi","family":"Yokota","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4256-9129","authenticated-orcid":false,"given":"Shin-ichi","family":"Horike","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shigehiro","family":"Karashima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isamu","family":"Miyamori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masakazu","family":"Yamagishi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshiyu","family":"Takeda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,9,11]]},"reference":[{"key":"11435_CR1","doi-asserted-by":"publisher","first-page":"2807","DOI":"10.1210\/jc.2015-1818","volume":"100","author":"LK Nieman","year":"2015","unstructured":"Nieman, L. K. et al. Treatment of Cushing\u2019s Syndrome: An Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. \n                           100, 2807\u20132831 (2015).","journal-title":"J Clin Endocrinol Metab."},{"key":"11435_CR2","doi-asserted-by":"publisher","first-page":"1526","DOI":"10.1210\/jc.2008-0125","volume":"93","author":"LK Nieman","year":"2008","unstructured":"Nieman, L. K. et al. The diagnosis of Cushing\u2019s syndrome: an Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. \n                           93, 1526\u20131540 (2008).","journal-title":"J Clin Endocrinol Metab."},{"key":"11435_CR3","doi-asserted-by":"publisher","first-page":"166","DOI":"10.1038\/ng1501","volume":"37","author":"LA Metherell","year":"2005","unstructured":"Metherell, L. A. et al. Mutations in MRAP, encoding a new interacting partner of the ACTH receptor, cause familial glucocorticoid deficiency type 2. Nat Genet. \n                           37, 166\u2013170 (2005).","journal-title":"Nat Genet."},{"key":"11435_CR4","doi-asserted-by":"publisher","first-page":"1019","DOI":"10.1056\/NEJMoa1310359","volume":"370","author":"F Beuschlein","year":"2014","unstructured":"Beuschlein, F. et al. Constitutive activation of PKA catalytic subunit in adrenal Cushing\u2019s syndrome. N Engl J Med. \n                           370, 1019\u20131028 (2014).","journal-title":"N Engl J Med."},{"key":"11435_CR5","doi-asserted-by":"publisher","first-page":"917","DOI":"10.1126\/science.1252328","volume":"344","author":"Y Sato","year":"2014","unstructured":"Sato, Y. et al. Recurrent somatic mutations underlie corticotropin-independent Cushing\u2019s syndrome. Science. \n                           344, 917\u2013920 (2014).","journal-title":"Science."},{"key":"11435_CR6","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1016\/0092-8674(92)90526-I","volume":"70","author":"A Bird","year":"1992","unstructured":"Bird, A. The essentials of DNA methylation. Cell. \n                           70, 5\u20138 (1992).","journal-title":"Cell."},{"key":"11435_CR7","doi-asserted-by":"publisher","first-page":"E536","DOI":"10.1210\/jc.2013-3495","volume":"99","author":"B Howard","year":"2014","unstructured":"Howard, B. et al. Integrated analysis of genome-wide methylation and gene expression shows epigenetic regulation of CYP11B2 in aldosteronomas. J Clin Endocrinol Metab. \n                           99, E536\u2013543 (2014).","journal-title":"J Clin Endocrinol Metab."},{"key":"11435_CR8","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1530\/EJE-15-0148","volume":"173","author":"M Murakami","year":"2015","unstructured":"Murakami, M. et al. Integration of transcriptome and methylome analysis of aldosterone-producing adenomas. Eur J Endocrinol. \n                           173, 185\u2013195 (2015).","journal-title":"Eur J Endocrinol."},{"key":"11435_CR9","doi-asserted-by":"publisher","first-page":"1432","DOI":"10.1161\/HYPERTENSIONAHA.116.08313","volume":"68","author":"Y Yoshii","year":"2016","unstructured":"Yoshii, Y. et al. Hypomethylation of CYP11B2 in Aldosterone-Producing Adenoma. Hypertension. \n                           68, 1432\u20131437 (2016).","journal-title":"Hypertension."},{"key":"11435_CR10","doi-asserted-by":"crossref","first-page":"20961","DOI":"10.1016\/S0021-9258(19)30030-4","volume":"264","author":"E Mornet","year":"1989","unstructured":"Mornet, E., Dupont, J., Vitek, A. & White, P. C. Characterization of two genes encoding human steroid 11 beta-hydroxylase P-450(11) beta. J Biol Chem. \n                           264, 20961\u201320967 (1989).","journal-title":"J Biol Chem."},{"key":"11435_CR11","doi-asserted-by":"publisher","first-page":"2296","DOI":"10.1210\/jc.2009-2010","volume":"95","author":"K Nishimoto","year":"2010","unstructured":"Nishimoto, K. et al. Adrenocortical zonation in humans under normal and pathological conditions. J Clin Endocrinol Metab. \n                           95, 2296\u20132305 (2010).","journal-title":"J Clin Endocrinol Metab."},{"key":"11435_CR12","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1016\/j.humpath.2016.03.016","volume":"54","author":"F Kubota-Nakayama","year":"2016","unstructured":"Kubota-Nakayama, F. et al. Expression of steroidogenic enzymes and their transcription factors in cortisol-producing adrenocortical adenomas: immunohistochemical analysis and quantitative real-time polymerase chain reaction studies. Hum Pathol. \n                           54, 165\u2013173 (2016).","journal-title":"Hum Pathol."},{"key":"11435_CR13","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1038\/hr.2015.42","volume":"38","author":"M Demura","year":"2015","unstructured":"Demura, M., Demura, Y., Takeda, Y. & Saijoh, K. Dynamic regulation of the angiotensinogen gene by DNA methylation, which is influenced by various stimuli experienced in daily life. Hypertens Res. \n                           38, 519\u2013527 (2015).","journal-title":"Hypertens Res."},{"key":"11435_CR14","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1016\/j.steroids.2011.10.010","volume":"77","author":"LC Cheng","year":"2012","unstructured":"Cheng, L. C., Pai, T. W. & Li, L. A. Regulation of human CYP11B1 and CYP11B2 promoters by transposable elements and conserved cis elements. Steroids. \n                           77, 100\u2013109 (2012).","journal-title":"Steroids."},{"key":"11435_CR15","doi-asserted-by":"publisher","first-page":"3587","DOI":"10.1210\/endo.141.10.7689","volume":"141","author":"XL Wang","year":"2000","unstructured":"Wang, X. L. et al. Transcriptional regulation of human 11beta-hydroxylase (hCYP11B1). Endocrinology. \n                           141, 3587\u20133594 (2000).","journal-title":"Endocrinology."},{"key":"11435_CR16","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1530\/EJE-14-1113","volume":"172","author":"A Thiel","year":"2015","unstructured":"Thiel, A. et al. PRKACA mutations in cortisol-producing adenomas and adrenal hyperplasia: a single-center study of 60 cases. Eur J Endocrinol. \n                           172, 677\u2013685 (2015).","journal-title":"Eur J Endocrinol."},{"key":"11435_CR17","doi-asserted-by":"publisher","first-page":"613","DOI":"10.1038\/ng.2956","volume":"46","author":"G Goh","year":"2014","unstructured":"Goh, G. et al. Recurrent activating mutation in PRKACA in cortisol-producing adrenal tumors. Nat Genet. \n                           46, 613\u2013617 (2014).","journal-title":"Nat Genet."},{"key":"11435_CR18","doi-asserted-by":"publisher","first-page":"768","DOI":"10.1126\/science.1198785","volume":"331","author":"M Choi","year":"2011","unstructured":"Choi, M. et al. K\u2009+\u2009channel mutations in adrenal aldosterone-producing adenomas and hereditary hypertension. Science. \n                           331, 768\u2013772 (2011).","journal-title":"Science."},{"key":"11435_CR19","doi-asserted-by":"publisher","first-page":"440","DOI":"10.1038\/ng.2550","volume":"45","author":"F Beuschlein","year":"2013","unstructured":"Beuschlein, F. et al. Somatic mutations in ATP1A1 and ATP2B3 lead to aldosterone-producing adenomas and secondary hypertension. Nat Genet. \n                           45, 440\u2013444 (2013).","journal-title":"Nat Genet."},{"key":"11435_CR20","doi-asserted-by":"publisher","first-page":"1050","DOI":"10.1038\/ng.2695","volume":"45","author":"UI Scholl","year":"2013","unstructured":"Scholl, U. I. et al. Somatic and germline CACNA1D calcium channel mutations in aldosterone-producing adenomas and primary aldosteronism. Nat Genet. \n                           45, 1050\u20131054 (2013).","journal-title":"Nat Genet."},{"key":"11435_CR21","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1038\/nature10716","volume":"480","author":"MB Stadler","year":"2011","unstructured":"Stadler, M. B. et al. DNA-binding factors shape the mouse methylome at distal regulatory regions. Nature. \n                           480, 490\u2013495 (2011).","journal-title":"Nature."},{"key":"11435_CR22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1003994","volume":"9","author":"A Feldmann","year":"2013","unstructured":"Feldmann, A. et al. Transcription factor occupancy can mediate active turnover of DNA methylation at regulatory regions. PLoS Genet. \n                           9, e1003994 (2013).","journal-title":"PLoS Genet."},{"key":"11435_CR23","doi-asserted-by":"publisher","first-page":"34287","DOI":"10.1074\/jbc.R113.512517","volume":"288","author":"A Blattler","year":"2013","unstructured":"Blattler, A. & Farnham, P. J. Cross-talk between site-specific transcription factors and DNA methylation states. J Biol Chem. \n                           288, 34287\u201334294 (2013).","journal-title":"J Biol Chem."},{"key":"11435_CR24","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1016\/j.devcel.2013.07.016","volume":"26","author":"BD Freedman","year":"2013","unstructured":"Freedman, B. D. et al. Adrenocortical zonation results from lineage conversion of differentiated zona glomerulosa cells. Dev Cell. \n                           26, 666\u2013673 (2013).","journal-title":"Dev Cell."},{"key":"11435_CR25","first-page":"526","volume":"31","author":"EA Kazerooni","year":"1990","unstructured":"Kazerooni, E. A. et al. Diagnostic accuracy and pitfalls of [iodine-131]6-beta-iodomethyl-19-norcholesterol (NP-59) imaging. J Nucl Med. \n                           31, 526\u2013534 (1990).","journal-title":"J Nucl Med."},{"key":"11435_CR26","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.mce.2013.11.022","volume":"383","author":"CE Gomez-Sanchez","year":"2014","unstructured":"Gomez-Sanchez, C. E. et al. Development of monoclonal antibodies against human CYP11B1 and CYP11B2. Mol Cell Endocrinol. \n                           383, 111\u2013117 (2014).","journal-title":"Mol Cell Endocrinol."},{"key":"11435_CR27","doi-asserted-by":"publisher","first-page":"22460","DOI":"10.1074\/jbc.M115.675595","volume":"290","author":"T Akagi","year":"2015","unstructured":"Akagi, T. et al. ETS-related transcription factors ETV4 and ETV5 are involved in proliferation and induction of differentiation-associated genes in embryonic stem (ES) cells. J Biol Chem. \n                           290, 22460\u201322473 (2015).","journal-title":"J Biol Chem."},{"key":"11435_CR28","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1016\/j.mce.2016.07.031","volume":"441","author":"A Tamura","year":"2017","unstructured":"Tamura, A. et al. Somatic KCNJ5 mutation occurring early in adrenal development may cause a novel form of juvenile primary aldosteronism. Mol Cell Endocrinol. \n                           441, 134\u2013139 (2017).","journal-title":"Mol Cell Endocrinol."},{"key":"11435_CR29","doi-asserted-by":"publisher","first-page":"1774","DOI":"10.1210\/en.2011-1733","volume":"153","author":"K Oki","year":"2012","unstructured":"Oki, K., Plonczynski, M. W., Luis Lam, M., Gomez-Sanchez, E. P. & Gomez-Sanchez, C. E. Potassium channel mutant KCNJ5 T158A expression in HAC-15 cells increases aldosterone synthesis. Endocrinology. \n                           153, 1774\u20131782 (2012).","journal-title":"Endocrinology."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-11435-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-11435-2","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-11435-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T10:45:51Z","timestamp":1671792351000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-11435-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,9,11]]},"references-count":29,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["11435"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-11435-2","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,9,11]]},"assertion":[{"value":"23 March 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 August 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 September 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"11205"}}