{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T15:08:02Z","timestamp":1787238482298,"version":"build-2736575974"},"reference-count":42,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,6,13]],"date-time":"2025-06-13T00:00:00Z","timestamp":1749772800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Promotion and Mutual Aid Corporation for Private Schools of Japan","award":["S1511008L"],"award-info":[{"award-number":["S1511008L"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Epigenomes"],"abstract":"<jats:p>Background: Somatic symptom disorder (SSD) in children may be influenced by stress reactivity and psychosocial factors. The glucocorticoid receptor (GR), encoded by NR3C1, is a key mediator of stress responses. However, the relationship between NR3C1 methylation and SSD remains unclear. Methods: We analyzed NR3C1 exon 1F methylation in cell-free DNA from saliva in 34 children with SSD and 29 age- and sex-matched controls using bisulfite amplicon sequencing. Psychological assessments included the Beck Depression Inventory-II (BDI-II) and KINDL questionnaires to evaluate associations with methylation patterns. Results: Methylation levels showed age-related differences. In children under 13, CpG sites displayed mixed methylation, and specific sites correlated with KINDL and BDI-II scores. KINDL physical and total well-being scores negatively correlated with CpG30 and positively with CpG35; BDI-II scores negatively correlated with CpG32 and CpG35. In children aged 13 or older, CpG sites showed uniformly high methylation with no correlation to psychological measures. The SSD group showed significantly higher average methylation across the exon 1F region than controls in the older age group. These children also had more cases of orthostatic dysregulation and longer illness duration. Conclusions: This study suggests age-dependent epigenetic regulation of NR3C1 in SSD. While younger children showed CpG-specific correlations with psychological symptoms, older children demonstrated uniformly high methylation and potentially reduced gene expression, potentially reflecting cumulative stress, autonomic dysfunction, and internalizing disorders such as anxiety and depression.<\/jats:p>","DOI":"10.3390\/epigenomes9020022","type":"journal-article","created":{"date-parts":[[2025,6,13]],"date-time":"2025-06-13T09:51:24Z","timestamp":1749808284000},"page":"22","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Methylation of the Glucocorticoid Receptor Gene in Children with Somatic Symptom Disorder: A Case-Control Study"],"prefix":"10.3390","volume":"9","author":[{"given":"Kyoko","family":"Hatta","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Juntendo University Faculty of Medicine, Tokyo 113-0033, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9392-3562","authenticated-orcid":false,"given":"Masato","family":"Kantake","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Juntendo University Faculty of Medicine, Tokyo 113-0033, Japan"},{"name":"Department of Neonatology, Juntendo University Nerima Hospital, 3-1-10 Nerima Takanodai, Nerima-ku, Tokyo 177-8521, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyoko","family":"Tanaka","sequence":"additional","affiliation":[{"name":"Department of Psychosocial Medicine, National Center for Child Health and Development, Tokyo 157-8535, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hirofumi","family":"Nakaoka","sequence":"additional","affiliation":[{"name":"Department of Cancer Genome Research, Sasaki Institute, Sasaki Foundation, Tokyo 101-0062, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0364-0022","authenticated-orcid":false,"given":"Toshiaki","family":"Shimizu","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Juntendo University Faculty of Medicine, Tokyo 113-0033, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2532-8910","authenticated-orcid":false,"given":"Hiromichi","family":"Shoji","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Juntendo University Faculty of Medicine, Tokyo 113-0033, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1111\/nmo.12741","article-title":"Genome-wide DNA methylation profiling of peripheral blood mononuclear cells in irritable bowel syndrome","volume":"28","author":"Mahurkar","year":"2016","journal-title":"Neurogastroenterol. Motil."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Eising, E., Datson, N.A., van den Maagdenberg, A.M.J.M., and Ferrari, M.D. (2013). Epigenetic mechanisms in migraine: A promising avenue?. BMC Med., 11.","DOI":"10.1186\/1741-7015-11-26"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1111\/j.1526-4610.2011.01906.x","article-title":"Childhood abuse and migraine: Epidemiology, sex differences, and potential mechanisms","volume":"51","author":"Tietjen","year":"2011","journal-title":"Headache"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1111\/j.1468-2982.2007.01303.x","article-title":"The triggers or precipitants of the acute migraine attack","volume":"27","author":"Kelman","year":"2007","journal-title":"Cephalalgia"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.neubiorev.2015.05.016","article-title":"Glucocorticoid receptor gene (NR3C1) methylation processes as mediators of early adversity in stress-related disorders causality: A critical review","volume":"55","author":"Leza","year":"2015","journal-title":"Neurosci. Biobehav. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1038\/nn1276","article-title":"Epigenetic programming by maternal behavior","volume":"7","author":"Weaver","year":"2004","journal-title":"Nat. Neurosci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1111\/cdev.12484","article-title":"Methylation of the Glucocorticoid Receptor Gene Promoter in Preschoolers: Links With Internalizing Behavior Problems","volume":"87","author":"Parade","year":"2016","journal-title":"Child. Dev."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Kantake, M., Ohkawa, N., Iwasaki, T., Ikeda, N., Awaji, A., Saito, N., Shoji, H., and Shimizu, T. (2018). Postnatal relative adrenal insufficiency results in methylation of the glucocorticoid receptor gene in preterm infants: A retrospective cohort study. Clin. Epigenetics, 10.","DOI":"10.1186\/s13148-018-0497-9"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1321","DOI":"10.4161\/epi.26634","article-title":"The roles of DNA methylation of NR3C1 and 11\u03b2-HSD2 and exposure to maternal mood disorder in utero on newborn neurobehavior","volume":"8","author":"Conradt","year":"2013","journal-title":"Epigenetics"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"97","DOI":"10.4161\/epi.3.2.6034","article-title":"Prenatal exposure to maternal depression, neonatal methylation of human glucocorticoid receptor gene (NR3C1) and infant cortisol stress responses","volume":"3","author":"Oberlander","year":"2008","journal-title":"Epigenetics"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1017\/S0954579415000176","article-title":"Methylation of exons 1D, 1F, and 1H of the glucocorticoid receptor gene promoter and exposure to adversity in preschool-aged children","volume":"27","author":"Tyrka","year":"2015","journal-title":"Dev. Psychopathol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Perroud, N., Paoloni-Giacobino, A., Prada, P., Oli\u00e9, E., Salzmann, A., Nicastro, R., Guillaume, S., Mouthon, D., Stouder, C., and Dieben, K. (2011). Increased methylation of glucocorticoid receptor gene (NR3C1) in adults with a history of childhood maltreatment: A link with the severity and type of trauma. Transl. Psychiatry, 1.","DOI":"10.1038\/tp.2011.60"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.1017\/S1461145713000102","article-title":"Genetic and epigenetic associations of MAOA and NR3C1 with depression and childhood adversities","volume":"16","author":"Melas","year":"2013","journal-title":"Int. J. Neuropsychopharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Sch\u00fcr, R.R., Boks, M.P., Rutten, B.P.F., Daskalakis, N.P., de Nijs, L., van Zuiden, M., Kavelaars, A., Heijnen, C.J., Jo\u00ebls, M., and Kahn, R.S. (2017). Longitudinal changes in glucocorticoid receptor exon 1F methylation and psychopathology after military deployment. Transl. Psychiatry, 7.","DOI":"10.1038\/tp.2017.150"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1101\/gad.2037511","article-title":"CpG islands and the regulation of transcription","volume":"25","author":"Deaton","year":"2011","journal-title":"Genes. Dev."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"American Psychiatric Association (2013). Diagnostic and Statistical Manual of Mental Disorders, American Psychiatric Publishing. [5th ed.].","DOI":"10.1176\/appi.books.9780890425596"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1678","DOI":"10.1111\/dme.13779","article-title":"WHO-5 and BDI-II are acceptable screening instruments for depression in people with diabetes","volume":"35","author":"Rauwerda","year":"2018","journal-title":"Diabet. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1590\/1516-4446-2012-1048","article-title":"Psychometric properties of the Beck Depression Inventory-II: A comprehensive review","volume":"35","author":"Wang","year":"2013","journal-title":"Rev. Bras. Psiquiatr."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S0165-1781(02)00106-3","article-title":"Cross-cultural validation of the Beck Depression Inventory-II in Japan","volume":"110","author":"Kojima","year":"2002","journal-title":"Psychiatry Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1007\/s10578-015-0568-5","article-title":"Cross-cultural measurement equivalence of the KINDL questionnaire for quality of life assessment in children and adolescents","volume":"47","author":"Jafari","year":"2016","journal-title":"Child. Psychiatry Hum. Dev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"187","DOI":"10.2332\/allergolint.08-OA-0034","article-title":"Exercise-induced asthma is associated with impaired quality of life among children with asthma in Japan","volume":"58","author":"Kojima","year":"2009","journal-title":"Allergol. Int."},{"key":"ref_22","first-page":"1404","article-title":"A study of the Kiddo-KINDL (Questionnaire for Measuring Health-Related Quality of Life in Children, Revised Version for 13 to 17-year-olds) in Japan","volume":"111","author":"Matsuzaki","year":"2007","journal-title":"J. Jpn. Pediatr. Soc."},{"key":"ref_23","first-page":"1514","article-title":"A study of the Kid-KINDL questionnaire for measuring quality of life in elementary school children in Japan","volume":"107","author":"Shibata","year":"2003","journal-title":"J. Jpn. Pediatr. Soc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1007\/s10519-012-9579-1","article-title":"Comparison of whole-genome DNA methylation patterns in whole blood, saliva, and lymphoblastoid cell lines","volume":"43","author":"Thompson","year":"2013","journal-title":"Behav. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Nazzari, S., Grumi, S., Mambretti, F., Villa, M., Giorda, R., and Provenzi, L. (2022). Maternal and infant NR3C1 and SLC6A4 epigenetic signatures of the COVID-19 pandemic lockdown: When timing matters. Transl. Psychiatry., 12.","DOI":"10.1038\/s41398-022-02160-0"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Leakey, T.I., Zielinski, J., Siegfried, R.N., Siegel, E.R., Fan, C.Y., and Cooney, C.A. (2008). A simple algorithm for quantifying DNA methylation levels on multiple independent CpG sites in bisulfite genomic sequencing electropherograms. Nucleic Acids Res., 36.","DOI":"10.1093\/nar\/gkn210"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.pnpbp.2018.09.002","article-title":"Epigenetics-by-sex interaction for somatization conferred by methylation at the promoter region of SLC6A4 gene","volume":"89","author":"Peralta","year":"2019","journal-title":"Prog. Neuropsychopharmacol. Biol. Psychiatry."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"376","DOI":"10.3748\/wjg.v20.i2.376","article-title":"Molecular basis of the irritable bowel syndrome","volume":"20","author":"Vaiopoulou","year":"2014","journal-title":"World J. Gastroenterol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.neubiorev.2018.08.017","article-title":"Glucocorticoid receptor gene (NR3C1) DNA methylation in association with trauma, psychopathology, transcript expression, or genotypic variation: A systematic review","volume":"95","author":"Watkeys","year":"2018","journal-title":"Neurosci. Biobehav. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Tyrka, A.R., Parade, S.H., Welch, E.S., Ridout, K.K., Price, L.H., Marsit, C., Philip, N.S., and Carpenter, L.L. (2016). Methylation of the leukocyte glucocorticoid receptor gene promoter in adults: Associations with early adversity and depressive, anxiety and substance-use disorders. Transl. Psychiatry, 6.","DOI":"10.1038\/tp.2016.112"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/S0304-3959(00)00483-8","article-title":"A cross-national study of the course of persistent pain in primary care","volume":"92","author":"Gureje","year":"2001","journal-title":"Pain"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1093\/jpepsy\/20.2.233","article-title":"Long-term health outcomes in patients with recurrent abdominal pain","volume":"20","author":"Walker","year":"1995","journal-title":"J. Pediatr. Psychol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1037\/0022-006X.69.1.85","article-title":"The relation of daily stressors to somatic and emotional symptoms in children with and without recurrent abdominal pain","volume":"69","author":"Walker","year":"2001","journal-title":"J. Consult. Clin. Psychol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Kozlowska, K., Scher, S., and Helgeland, H. (2020). Functional Somatic Symptoms in Children and Adolescents: A Stress-System Approach to Assessment and Treatment, Springer Nature.","DOI":"10.1007\/978-3-030-46184-3"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1016\/j.bbi.2020.12.018","article-title":"Linking the hemodynamic consequences of adverse childhood experiences to an altered HPA axis and acute stress response","volume":"93","author":"Dempster","year":"2021","journal-title":"Brain Behav. Immun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1097\/PSY.0000000000001237","article-title":"DNA methylation signatures of functional somatic syndromes: Systematic review","volume":"85","author":"Fischer","year":"2023","journal-title":"Psychosom. Med."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Emich-Widera, E., Kazek, B., Szwed-Bialozyt, B., Kopyta, I., and Kostorz, A. (2012). Headaches as somatoform disorders in children and adolescents. Ment. Illn., 4.","DOI":"10.4081\/mi.2012.e9"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Guidetti, V., Faedda, N., and Siniatchkin, M. (2016). Migraine in childhood: Biobehavioural or psychosomatic disorder?. J. Headache Pain, 17.","DOI":"10.1186\/s10194-016-0675-0"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1007\/s13311-013-0213-6","article-title":"Epigenetics and psychiatry","volume":"10","author":"Mahgoub","year":"2013","journal-title":"Neurotherapeutics"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"616","DOI":"10.3748\/wjg.v18.i7.616","article-title":"Psychosocial determinants of irritable bowel syndrome","volume":"18","author":"Baban","year":"2012","journal-title":"World J. Gastroenterol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5532","DOI":"10.3748\/wjg.v21.i18.5532","article-title":"Psychosocial mechanisms for the transmission of somatic symptoms from parents to children","volume":"21","author":"Levy","year":"2015","journal-title":"World J. Gastroenterol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1111\/j.1572-0241.2007.01722.x","article-title":"Early life risk factors that contribute to irritable bowel syndrome in adults: A systematic review","volume":"103","author":"Chitkara","year":"2008","journal-title":"Am. J. Gastroenterol."}],"container-title":["Epigenomes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4655\/9\/2\/22\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:51:31Z","timestamp":1760032291000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4655\/9\/2\/22"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,13]]},"references-count":42,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["epigenomes9020022"],"URL":"https:\/\/doi.org\/10.3390\/epigenomes9020022","relation":{},"ISSN":["2075-4655"],"issn-type":[{"value":"2075-4655","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,13]]}}}