{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T05:41:37Z","timestamp":1762321297640,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T00:00:00Z","timestamp":1762128000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008891","name":"Loma Linda University","doi-asserted-by":"crossref","award":["2200459"],"award-info":[{"award-number":["2200459"]}],"id":[{"id":"10.13039\/100008891","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Glucocorticoids are essential for fetal organ maturation and form the basis of antenatal corticosteroid therapy that has significantly reduced preterm-related morbidity such as respiratory distress syndrome (RDS). However, neonatal morbidity remains a clinical challenge regardless of antenatal corticosteroid therapy. Currently, it is thought that adverse intrauterine environments dysregulate glucocorticoid receptor (GR) homeostasis, yet the biological mechanisms remain poorly understood. Therefore, we aimed to study ex vivo glucocorticoid sensitivity in cord blood immune cells from two independent preterm cohorts to identify associations with neonatal morbidity and uncover potential mechanisms of dysregulated glucocorticoid homeostasis. In the first cohort, thawed cord blood mononuclear cells were exposed to betamethasone in the presence of lipopolysaccharides (LPS) for 4 h. In the second cohort, freshly isolated white blood cells were treated with dexamethasone under unstimulated and LPS-stimulated conditions for 48 h. GR isoform expression and regulation of transactivated and transrepressed genes were assessed via qPCR, immunoblotting, flow cytometry, and ELISA. In both cohorts, reduced GR expression, particularly of the GR\u03b1 isoform, was observed in neonates with morbidity, but only with culture time and not in freshly isolated cells. Ex vivo impaired glucocorticoid-mediated transrepression of proinflammatory genes IL6 and TNF was also observed in the morbidity groups. In contrast, all samples were comparable in basal immune cell distributions and transactivation of glucocorticoid response element (GRE)-dependent genes GILZ and FKBP5, irrespective of neonatal morbidity. These findings suggest that neonates that develop morbidities experience an early postnatal GR dysfunction that is potentially programmed in utero. Moreover, under conditions of decreased GR abundance, classical transactivation functions appear to be preserved at the expense of more complex regulatory mechanisms such as transrepression.<\/jats:p>","DOI":"10.3390\/ijms262110686","type":"journal-article","created":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T13:15:14Z","timestamp":1762175714000},"page":"10686","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Decreased Glucocorticoid Receptor Expression and Function in Cord Blood Immune Cells from Preterm Neonates with Morbidity"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-8473-190X","authenticated-orcid":false,"given":"Nana A. O.","family":"Anti","sequence":"first","affiliation":[{"name":"Lawrence D. Longo MD Center for Perinatal Biology, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Douglas D.","family":"Deming","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Division of Neonatology, Loma Linda University Children\u2019s Hospital, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ciprian P.","family":"Gheorghe","sequence":"additional","affiliation":[{"name":"Lawrence D. Longo MD Center for Perinatal Biology, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92354, USA"},{"name":"Department of Gynecology and Obstetrics, Division of Maternal-Fetal Medicine, Loma Linda University Children\u2019s Hospital, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashra","family":"Tugung","sequence":"additional","affiliation":[{"name":"Department of Gynecology and Obstetrics, Division of Maternal-Fetal Medicine, Loma Linda University Children\u2019s Hospital, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikia","family":"Gray-Hutto","sequence":"additional","affiliation":[{"name":"Department of Gynecology and Obstetrics, Division of Maternal-Fetal Medicine, Loma Linda University Children\u2019s Hospital, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9386-8938","authenticated-orcid":false,"given":"Lubo","family":"Zhang","sequence":"additional","affiliation":[{"name":"Lawrence D. Longo MD Center for Perinatal Biology, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eugenia","family":"Mata-Greenwood","sequence":"additional","affiliation":[{"name":"Lawrence D. Longo MD Center for Perinatal Biology, Department of Basic Sciences, School of Medicine, Loma Linda University, Loma Linda, CA 92354, USA"},{"name":"Department of Gynecology and Obstetrics, Division of Maternal-Fetal Medicine, Loma Linda University Children\u2019s Hospital, Loma Linda, CA 92354, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Anti, N.A.O., Gheorghe, C.P., Deming, D.D., Adeoye, O.O., Zhang, L., and Mata-Greenwood, E. (2025). Perinatal Glucocorticoid Sensitivity in the Preterm Newborn: Molecular Mechanisms, Endogenous Determinants, and Clinical Implications. Front. Endocrinol., 16.","DOI":"10.3389\/fendo.2025.1587891"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/bs.ctdb.2016.12.004","article-title":"Mechanisms of Glucocorticoid Action During Development","volume":"125","author":"Busada","year":"2017","journal-title":"Curr. Top. Dev. Biol."},{"key":"ref_3","first-page":"CD004454","article-title":"Antenatal Corticosteroids for Accelerating Fetal Lung Maturation for Women at Risk of Preterm Birth","volume":"2021","author":"McGoldrick","year":"2020","journal-title":"Cochrane Database Syst. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1016\/j.ajog.2022.07.030","article-title":"The Complex Challenge of Antenatal Steroid Therapy Nonresponsiveness","volume":"227","author":"Takahashi","year":"2022","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1210\/endrev\/bnae008","article-title":"The Glucocorticoid Receptor: Isoforms, Functions, and Contribution to Glucocorticoid Sensitivity","volume":"45","author":"Lockett","year":"2024","journal-title":"Endocr. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5718","DOI":"10.1111\/febs.16100","article-title":"A Guide to Changing Paradigms of Glucocorticoid Receptor Function\u2014A Model System for Genome Regulation and Physiology","volume":"289","author":"Vettorazzi","year":"2022","journal-title":"FEBS J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1210\/er.2014-1010","article-title":"Comprehensive Overview of the Structure and Regulation of the Glucocorticoid Receptor","volume":"35","author":"Vandevyver","year":"2014","journal-title":"Endocr. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1038\/s41467-018-03780-1","article-title":"Cryptic Glucocorticoid Receptor-Binding Sites Pervade Genomic NF-\u03baB Response Elements","volume":"9","author":"Hudson","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8596","DOI":"10.1093\/nar\/gkx509","article-title":"Tethering Not Required: The Glucocorticoid Receptor Binds Directly to Activator Protein-1 Recognition Motifs to Repress Inflammatory Genes","volume":"45","author":"Weikum","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Gerber, A.N., Newton, R., and Sasse, S.K. (2021). Repression of Transcription by the Glucocorticoid Receptor: A Parsimonious Model for the Genomics Era. J. Biol. Chem., 296.","DOI":"10.1016\/j.jbc.2021.100687"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1038\/nrendo.2013.183","article-title":"Glucocorticoid Sensitivity in Health and Disease","volume":"9","author":"Quax","year":"2013","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"R328","DOI":"10.1530\/EC-18-0421","article-title":"Disease- and Treatment-associated Acquired Glucocorticoid Resistance","volume":"7","author":"Wilkinson","year":"2018","journal-title":"Endocr. Connect."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1172\/JCI107581","article-title":"Cytoplasmic Receptor for Glucocorticoids in Lung of the Human Fetus and Neonate","volume":"53","author":"Ballard","year":"1974","journal-title":"J. Clin. Investig."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/0022-4731(85)90106-2","article-title":"Low Levels of Glucocorticoid Binding Sites in Circulating Lymphocytes of Premature Infants Suffering from Hyaline Membrane Disease","volume":"22","author":"Kerepesi","year":"1985","journal-title":"J. Steroid. Biochem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1080\/14767058.2016.1174992","article-title":"Clinical Significance of Glucocorticoid Receptor Expression in Premature Infants with Respiratory Distress Syndrome","volume":"30","author":"Ataseven","year":"2017","journal-title":"J. Matern. Fetal. Neonatal Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1350","DOI":"10.1111\/j.1651-2227.1997.tb14912.x","article-title":"Glucocorticoid Receptors in Cord Blood Lymphocytes of Healthy Neonates and of Preterms Suffering from Respiratory Distress Syndrome","volume":"86","author":"Lentjes","year":"1997","journal-title":"Acta Paediatr."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1515\/jpem-2012-0170","article-title":"Glucocorticoid Receptor Expression in Whole Blood with Preterm Infants","volume":"26","author":"Go","year":"2013","journal-title":"J. Pediatr. Endocrinol. Metab."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1159\/000526915","article-title":"Glucocorticoid Receptor Expression Pattern in Very Low-Birth Weight Infants Changes Drastically within the First Week of Life","volume":"96","author":"Yamamoto","year":"2022","journal-title":"Horm. Res. Paediatr."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1159\/000014099","article-title":"Effects of Dexamethasone on IL-1\u03b2, IL-6, and TNF-\u03b1 Production by Mononuclear Cells of Newborns and Adults","volume":"75","author":"Bessler","year":"1999","journal-title":"Neonatology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1634","DOI":"10.1038\/s41372-020-00788-8","article-title":"Novel Biomarkers of Bronchopulmonary Dysplasia and Bronchopulmonary Dysplasia-Associated Pulmonary Hypertension","volume":"40","author":"Sahni","year":"2020","journal-title":"J. Perinatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"F198","DOI":"10.1136\/fn.77.3.F198","article-title":"Early Increase of TNF alpha and IL-6 in Tracheobronchial Aspirate Fluid Indicator of Subsequent Chronic Lung Disease in Preterm Infants","volume":"77","author":"Jonsson","year":"1997","journal-title":"Arch. Dis. Child.-Fetal Neonatal Ed."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1542\/peds.102.4.927","article-title":"Early Clinical Markers for the Development of Bronchopulmonary Dysplasia: Soluble E-Selectin and ICAM-1","volume":"102","author":"Ramsay","year":"1998","journal-title":"Pediatrics"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Martin, C.B., Oshiro, B.T., Sands, L.D., Kabir, S., Thorpe, D., Clark, T.C., Yao, R., and Mata-Greenwood, E. (2020). Vitamin-D Dysregulation in Early- and Late-Onset Preeclampsia: A Gestational-Age Matched Study. J. Steroid Biochem. Mol. Biol., 203.","DOI":"10.1016\/j.jsbmb.2020.105729"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0303-7207(01)00722-5","article-title":"The Anti-Inflammatory Action of Glucocorticoids Is Mediated by Cell Type Specific Regulation of Apoptosis","volume":"189","author":"Amsterdam","year":"2002","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1113\/jphysiol.2007.136796","article-title":"Identification of Glucocorticoid-Regulated Genes That Control Cell Proliferation during Murine Respiratory Development","volume":"585","author":"Bird","year":"2007","journal-title":"J. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.4049\/jimmunol.169.4.1837","article-title":"Thymocyte Apoptosis Induced by T Cell Activation Is Mediated by Glucocorticoids In Vivo","volume":"169","author":"Brewer","year":"2002","journal-title":"J. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1101\/gad.9.13.1608","article-title":"Targeted Disruption of the Glucocorticoid Receptor Gene Blocks Adrenergic Chromaffin Cell Development and Severely Retards Lung Maturation","volume":"9","author":"Cole","year":"1995","journal-title":"Genes Dev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1165\/rcmb.2003-0236OC","article-title":"Altered Epithelial Cell Proportions in the Fetal Lung of Glucocorticoid Receptor Null Mice","volume":"30","author":"Cole","year":"2004","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Kokkinopoulou, I., and Moutsatsou, P. (2021). Mitochondrial Glucocorticoid Receptors and Their Actions. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22116054"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.freeradbiomed.2019.04.029","article-title":"Role of oxidative stress in neonatal respiratory distress syndrome","volume":"142","author":"Marseglia","year":"2019","journal-title":"Free Rad. Biol. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/S0092-8674(00)81183-6","article-title":"DNA binding of the glucocorticoid receptor is not essential for survival","volume":"93","author":"Reichardt","year":"1998","journal-title":"Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/pr.2014.54","article-title":"Administration of Antenatal Glucocorticoids and Postnatal Surfactant Ameliorates Respiratory Distress Syndrome\u2013Associated Neonatal Lethality in Erk3\u2212\/\u2212 Mouse Pups","volume":"76","author":"Sbrana","year":"2014","journal-title":"Pediatr. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1280","DOI":"10.1210\/me.2009-0380","article-title":"Glucocorticoid Activity during Lung Maturation Is Essential in Mesenchymal and Less in Alveolar Epithelial Cells","volume":"25","author":"Habermehl","year":"2011","journal-title":"Mol. Endocrinol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Li, A., Hardy, R., Stoner, S., Tuckermann, J., Seibel, M., and Zhou, H. (2013). Deletion of Mesenchymal Glucocorticoid Receptor Attenuates Embryonic Lung Development and Abdominal Wall Closure. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0063578"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e139452","DOI":"10.1172\/jci.insight.139452","article-title":"Prenatal Inflammation Enhances Antenatal Corticosteroid\u2013Induced Fetal Lung Maturation","volume":"5","author":"Schmidt","year":"2020","journal-title":"JCI Insight"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1808","DOI":"10.1080\/21505594.2021.1948276","article-title":"Transcriptome Analysis of Porcine PBMCs Reveals Lipopolysaccharide-Induced Immunomodulatory Responses and Crosstalk of Immune and Glucocorticoid Receptor Signaling","volume":"12","author":"Li","year":"2021","journal-title":"Virulence"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1008","DOI":"10.1006\/bbrc.2000.3079","article-title":"Glucocorticoid Effects on NF-\u03baB Binding in the Transcription of the ICAM-1 Gene","volume":"273","author":"Liden","year":"2000","journal-title":"Biochem. Biophy. Res. Commun."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2128","DOI":"10.4049\/jimmunol.163.4.2128","article-title":"Up-Regulation of ICAM-1 by Cytokines in Human Tracheal Smooth Muscle Cells Involves an NF-kB- Dependent Signaling Pathway That Is Only Partially Sensitive to Dexamethasone","volume":"163","author":"Amrani","year":"1999","journal-title":"J. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1542\/peds.111.3.461","article-title":"Soluble E-Selectin, Soluble L-Selectin and Soluble ICAM-1 in Bronchopulmonary Dysplasia, and Changes with Dexamethasone","volume":"111","author":"Ballabh","year":"2003","journal-title":"Pediatrics"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1148\/radiology.157.2.3840268","article-title":"Respiratory distress syndrome treated with human surfactant: Radiographic findings","volume":"157","author":"Edwards","year":"1985","journal-title":"Radiology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1016\/j.clp.2013.07.004","article-title":"Respiratory disorders in moderately preterm, late preterm, and early term infants","volume":"40","author":"Mahoney","year":"2013","journal-title":"Clin. Perinatol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1164\/rccm.201812-2348OC","article-title":"The Diagnosis of Bronchopulmonary Dysplasia in Very Preterm Infants: An Evidence-Based Approach","volume":"200","author":"Jensen","year":"2019","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1016\/S0022-3476(78)80282-0","article-title":"Incidence and evolution of subependymal and intraventricular hemorrhage: A study of infants with birth weights less than 1500 gm","volume":"92","author":"Papile","year":"1978","journal-title":"J. Pediatr."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1097\/00000658-197801000-00001","article-title":"Neonatal necrotizing enterocolitis. Therapeutic decisions based upon clinical staging","volume":"187","author":"Bell","year":"1978","journal-title":"Ann. Surg."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1038\/pr.2017.66","article-title":"Glucocorticoid receptor expression on circulating leukocytes in healthy and asthmatic adolescents in response to exercise","volume":"82","author":"Lu","year":"2017","journal-title":"Pediatr. Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1017\/cts.2016.20","article-title":"Glucocorticoid receptor expression on circulating leukocytes differs between healthy male and female adults","volume":"1","author":"Lu","year":"2017","journal-title":"J. Clin. Transl. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.jim.2010.03.006","article-title":"Comparative in vitro stimulation with lipopolysaccharide to study TNF alpha gene expression in fresh whole blood, fresh and frozen peripheral blood mononuclear cells","volume":"357","author":"Chen","year":"2010","journal-title":"J. Immunol. Methods"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1530\/eje.0.1500041","article-title":"A comparison of in vitro bioassays to determine cellular glucocorticoid sensitivity","volume":"150","author":"Vermeer","year":"2004","journal-title":"Eur. J. Endocrin."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1530\/JME-15-0124","article-title":"Endothelial glucocorticoid receptor promoter methylation according to dexamethasone sensitivity","volume":"55","author":"Jackson","year":"2015","journal-title":"J. Mol. Endocrin."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1046","DOI":"10.1161\/ATVBAHA.113.301247","article-title":"Role of BCL2-Associated Athanogene 1 in Differential Sensitivity of Human Endothelial Cells to Glucocorticoids","volume":"33","author":"Stewart","year":"2013","journal-title":"Arterioscler. Thromb. Vasc. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/21\/10686\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T05:28:40Z","timestamp":1762320520000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/21\/10686"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,3]]},"references-count":50,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ijms262110686"],"URL":"https:\/\/doi.org\/10.3390\/ijms262110686","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2025,11,3]]}}}