Association of DNA methylation with steroidogenic enzymes in Cushing’s adenoma

in Endocrine-Related Cancer
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  • 1 Department of Molecular and Internal Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan
  • | 2 Department of Urology, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan
  • | 3 Department of Anatomical Pathology, Hiroshima University Hospital, Hiroshima, Japan
  • | 4 Division of Endocrinology, G.V. (Sonny) Montgomery VA Medical Center and University of Mississippi Medical Center, Jackson, Mississippi, USA
  • | 5 Department of Pharmacology & Toxicology, University of Mississippi Medical Center, Jackson, Mississippi, USA

Contributor Notes

Correspondence should be addressed to K Oki: kenjioki@hiroshima-u.ac.jp
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DNA methylation and demethylation regulate the transcription of genes. DNA methylation-associated gene expression of adrenal steroidogenic enzymes may regulate cortisol production in cortisol-producing adenoma (CPA). We aimed to determine the DNA methylation levels of all genes encoding steroidogenic enzymes involved in CPA. Additionally, the aims were to clarify the DNA methylation-associated gene expression and evaluate the difference of CPA genotype from others using DNA methylation data. Twenty-five adrenal CPA and six nonfunctioning adrenocortical adenoma (NFA) samples were analyzed. RNA sequencing and DNA methylation array were performed. The methylation levels at 118 methylation sites of the genes were investigated, and their methylation and mRNA levels were subsequently integrated. Among all the steroidogenic enzyme genes studied, CYP17A1 gene was mainly found to be hypomethylated in CPA compared to that in NFA, and the Benjamini-Hochberg procedure demonstrated that methylation levels at two sites in the CYP17A1 gene body were statistically significant. PRKACA mutant CPAs predominantly exhibited hypomethylation of CYP17A1 gene compared with the GNAS mutant CPAs. Inverse associations between CYP17A1 methylation in three regions of the gene body and its mRNA levels were observed in the NFAs and CPAs. In applying clustering analysis using CYP17A1 methylation and mRNA levels, CPAs with PRKACA mutation were differentiated from NFAs and CPAs with a GNAS mutation. We demonstrated that CPAs exhibited hypomethylation of the CYP17A1 gene body in CPA, especially in the PRKACA mutant CPAs. Methylation of CYP17A1 gene may influence its transcription levels.

 

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  • Baba R, Oki K, Gomez-Sanchez CE, Otagaki Y, Itcho K, Kobuke K, Kodama T, Nagano G, Ohno H, Yoneda M, et al.2021 Genotype-specific cortisol production associated with Cushing’s syndrome adenoma with PRKACA mutations. Molecular and Cellular Endocrinology 538 111456. (https://doi.org/10.1016/j.mce.2021.111456)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Beuschlein F, Fassnacht M, Assie G, Calebiro D, Stratakis CA, Osswald A, Ronchi CL, Wieland T, Sbiera S, Faucz FR, et al.2014 Constitutive activation of PKA catalytic subunit in adrenal Cushing’s syndrome. New England Journal of Medicine 370 10191028. (https://doi.org/10.1056/NEJMoa1310359)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Brown SJ, Stoilov P & Xing Y 2012 Chromatin and epigenetic regulation of pre-mRNA processing. Human Molecular Genetics 21 R90R96. (https://doi.org/10.1093/hmg/dds353)

  • Calebiro D, Hannawacker A, Lyga S, Bathon K, Zabel U, Ronchi C, Beuschlein F, Reincke M, Lorenz K, Allolio B, et al.2014 PKA catalytic subunit mutations in adrenocortical Cushing’s adenoma impair association with the regulatory subunit. Nature Communications 5 5680. (https://doi.org/10.1038/ncomms6680)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Cohanim AB & Haran TE 2009 The coexistence of the nucleosome positioning code with the genetic code on eukaryotic genomes. Nucleic Acids Research 37 64666476. (https://doi.org/10.1093/nar/gkp689)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Di Dalmazi G, Altieri B, Scholz C, Sbiera S, Luconi M, Waldman J, Kastelan D, Ceccato F, Chiodini I, Arnaldi G, et al.2020a RNA sequencing and somatic mutation status of adrenocortical tumors: novel pathogenetic insights. Journal of Clinical Endocrinology and Metabolism 105 dgaa616. (https://doi.org/10.1210/clinem/dgaa616)

    • Search Google Scholar
    • Export Citation
  • Di Dalmazi G, Morandi L, Rubin B, Pilon C, Asioli S, Vicennati V, De Leo A, Ambrosi F, Santini D, Pagotto U, et al.2020b DNA methylation of steroidogenic enzymes in benign adrenocortical tumors: new insights in aldosterone-producing adenomas. Journal of Clinical Endocrinology and Metabolism 105 dgaa585. (https://doi.org/10.1210/clinem/dgaa585)

    • Search Google Scholar
    • Export Citation
  • Goh G, Scholl UI, Healy JM, Choi M, Prasad ML, Nelson-Williams C, Kunstman JW, Korah R, Suttorp AC, Dietrich D, et al.2014 Recurrent activating mutation in PRKACA in cortisol-producing adrenal tumors. Nature Genetics 46 613617. (https://doi.org/10.1038/ng.2956)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Gomez-Sanchez CE, Qi X, Velarde-Miranda C, Plonczynski MW, Parker CR, Rainey W, Satoh F, Maekawa T, Nakamura Y, Sasano H, et al.2014 Development of monoclonal antibodies against human CYP11B1 and CYP11B2. Molecular and Cellular Endocrinology 383 111117. (https://doi.org/10.1016/j.mce.2013.11.022)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Howard B, Wang Y, Xekouki P, Faucz FR, Jain M, Zhang L, Meltzer PG, Stratakis CA & Kebebew E 2014 Integrated analysis of genome-wide methylation and gene expression shows epigenetic regulation of CYP11B2 in aldosteronomas. Journal of Clinical Endocrinology and Metabolism 99 E536E543. (https://doi.org/10.1210/jc.2013-3495)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Hyeon JW, Kim AH & Yano H 2021 Epigenetic regulation in Huntington’s disease. Neurochemistry International 148 105074. (https://doi.org/10.1016/j.neuint.2021.105074)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Itcho K, Oki K, Gomez-Sanchez CE, Gomez-Sanchez EP, Ohno H, Kobuke K, Nagano G, Yoshii Y, Baba R, Hattori N, et al.2020 Endoplasmic reticulum chaperone calmegin is upregulated in aldosterone-producing adenoma and associates With aldosterone production. Hypertension 75 492499. (https://doi.org/10.1161/HYPERTENSIONAHA.119.14062)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Kan RL, Chen J & Sallam T 2021 Crosstalk between epitranscriptomic and epigenetic mechanisms in gene regulation. Trends in Genetics 38 182193. (https://doi.org/10.1016/j.tig.2021.06.014)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Klutstein M, Nejman D, Greenfield R & Cedar H 2016 DNA methylation in cancer and aging. Cancer Research 76 34463450. (https://doi.org/10.1158/0008-5472.CAN-15-3278)

  • Kobuke K, Oki K, Gomez-Sanchez CE, Ohno H, Itcho K, Yoshii Y, Yoneda M & Hattori N 2018 Purkinje cell protein 4 expression is associated With DNA methylation status in aldosterone-producing adenoma. Journal of Clinical Endocrinology and Metabolism 103 965971. (https://doi.org/10.1210/jc.2017-01996)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Kometani M, Yoneda T, Demura M, Koide H, Nishimoto K, Mukai K, Gomez-Sanchez CE, Akagi T, Yokota T, Horike SI, et al.2017 Cortisol overproduction results from DNA methylation of CYP11B1 in hypercortisolemia. Scientific Reports 7 11205. (https://doi.org/10.1038/s41598-017-11435-2)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Koralewski TE & Krutovsky KV 2011 Evolution of exon-intron structure and alternative splicing. PLoS ONE 6 e18055. (https://doi.org/10.1371/journal.pone.0018055)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Kubota-Nakayama F, Nakamura Y, Konosu-Fukaya S, Azmahani A, Ise K, Yamazaki Y, Kitawaki Y, Felizola SJ, Ono Y, Omata K, et al.2016 Expression of steroidogenic enzymes and their transcription factors in cortisol-producing adrenocortical adenomas: immunohistochemical analysis and quantitative real-time polymerase chain reaction studies. Human Pathology 54 165173. (https://doi.org/10.1016/j.humpath.2016.03.016)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • La Rosa S, Furlan D, Franzi F, Battaglia P, Frattini M, Zanellato E, Marando A, Sahnane N, Turri-Zanoni M, Castelnuovo P, et al.2013 Mixed exocrine-neuroendocrine carcinoma of the nasal cavity: clinico-pathologic and molecular study of a case and review of the literature. Head and Neck Pathology 7 7684. (https://doi.org/10.1007/s12105-012-0379-y)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Lacroix A, Feelders RA, Stratakis CA & Nieman LK 2015 Cushing’s syndrome. Lancet 386 913927. (https://doi.org/10.1016/S0140-6736(1461375-1)

  • Legendre CR, Demeure MJ, Whitsett TG, Gooden GC, Bussey KJ, Jung S, Waibhav T, Kim S & Salhia B 2016 Pathway implications of aberrant global methylation in adrenocortical cancer. PLoS ONE 11 e0150629. (https://doi.org/10.1371/journal.pone.0150629)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Lev Maor G, Yearim A & Ast G 2015 The alternative role of DNA methylation in splicing regulation. Trends in Genetics 31 274280. (https://doi.org/10.1016/j.tig.2015.03.002)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Martisova A, Holcakova J, Izadi N, Sebuyoya R, Hrstka R & Bartosik M 2021 DNA methylation in solid tumors: functions and methods of detection. International Journal of Molecular Sciences 22 4247. (https://doi.org/10.3390/ijms22084247)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Moran S, Arribas C & Esteller M 2016 Validation of a DNA methylation microarray for 850,000 CpG sites of the human genome enriched in enhancer sequences. Epigenomics 8 389399. (https://doi.org/10.2217/epi.15.114)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Nanao Y, Oki K, Kobuke K, Itcho K, Baba R, Kodama T, Otagaki Y, Okada A, Yoshii Y, Nagano G, et al.2022 Hypomethylation associated vitamin D receptor expression in ATP1A1 mutant aldosterone-producing adenoma. Molecular and Cellular Endocrinology 548 111613. (https://doi.org/10.1016/j.mce.2022.111613)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Nieman LK, Biller BM, Findling JW, Newell-Price J, Savage MO, Stewart PM & Montori VM 2008 The diagnosis of Cushing’s syndrome: an Endocrine Society clinical practice guideline. Journal of Clinical Endocrinology and Metabolism 93 15261540. (https://doi.org/10.1210/jc.2008-0125)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • Sakuma I, Suematsu S, Matsuzawa Y, Saito J, Omura M, Maekawa T, Nakamura Y, Sasano H & Nishikawa T 2013 Characterization of steroidogenic enzyme expression in aldosterone-producing adenoma: a comparison with various human adrenal tumors. Endocrine Journal 60 329336. (https://doi.org/10.1507/endocrj.ej12-0270)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Schwartz S, Meshorer E & Ast G 2009 Chromatin organization marks exon-intron structure. Nature Structural and Molecular Biology 16 990995. (https://doi.org/10.1038/nsmb.1659)

  • Slieker RC, Roost MS, van Iperen L, Suchiman HE, Tobi EW, Carlotti F, de Koning EJ, Slagboom PE, Heijmans BT & Chuva de Sousa Lopes SM 2015 DNA methylation landscapes of human fetal development. PLoS Genetics 11 e1005583. (https://doi.org/10.1371/journal.pgen.1005583)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Takeda Y, Demura M, Wang F, Karashima S, Yoneda T, Kometani M, Hashimoto A, Aono D, Horike SI, Meguro-Horike M, et al.2018 Epigenetic regulation of aldosterone synthase gene by sodium and angiotensin II. Journal of the American Heart Association 7 e008281. (https://doi.org/10.1161/JAHA.117.008281)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • Yoshii Y, Oki K, Gomez-Sanchez CE, Ohno H, Itcho K, Kobuke K & Yoneda M 2016 Hypomethylation of CYP11B2 in aldosterone-producing adenoma. Hypertension 68 14321437. (https://doi.org/10.1161/HYPERTENSIONAHA.116.08313)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Zardo G 2021 The role of H3K4 trimethylation in CpG islands hypermethylation in cancer. Biomolecules 11 143. (https://doi.org/10.3390/biom11020143)