title: Adrenal Steroid Metabolites Accumulating in Congenital Adrenal Hyperplasia Lead to Transactivation of the Glucocorticoid Receptor.
abstract: Patients with congenital adrenal hyperplasia (CAH) are often clinically less severely affected by cortisol deficiency than anticipated from their enzymatic defect. We hypothesize that adrenal steroid hormone precursors that accumulate in untreated or poorly controlled CAH have glucocorticoid activity and partially compensate for cortisol deficiency. We studied the in vitro effects of 17-hydroxyprogesterone (17OHP), progesterone (P), 21-deoxycortisol (21DF), and androstenedione (Δ4) on the human glucocorticoid receptor (hGR). Competitive binding assays were performed in HeLa cells. Nuclear translocation of the hGR was studied by transfection of COS-7 cells with a GFP-tagged hGR and fluorescence microscopy. Transactivation assays were performed in COS-7 cells and in HEK 293 cells after cotransfection with hGR and luciferase reporter vectors using a dual luciferase assay. 17OHP, P, and 21DF are able to bind to the hGR with binding affinities of 24-43% compared with cortisol. Δ4 has a low binding affinity. Incubation with 21DF led to complete nuclear translocation of the hGR, whereas treatment with 17OHP or P resulted in partial nuclear translocation. 21DF transactivated the hGR with an EC50 approximately 6 times the EC50 of cortisol. 17OHP and P transactivated the hGR with EC50s of more than 100 times the EC50 of cortisol. No hGR transactivation was detected after incubation with Δ4. 21DF, 17OHP, and P are able to bind, translocate, and transactivate the hGR in vitro and thus may have glucocorticoid activity. 21DF might have a clinically relevant agonistic effect on the hGR and could potentially partially compensate the cortisol deficiency in CAH patients.
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: K J
authors.initials: KJ
authors.lastname: Pijnenburg-Kleizen
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Engels
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: C F
authors.initials: CF
authors.lastname: Mooij
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Griffin
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: N
authors.initials: N
authors.lastname: Krone
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: P N
authors.initials: PN
authors.lastname: Span
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: A E
authors.initials: AE
authors.lastname: van Herwaarden
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: F C G J
authors.initials: FC
authors.lastname: Sweep
authors.affiliation: Department of Pediatrics (K.J.P.-K., M.E., C.F.M., H.L.C.v.d.G.), and Department of Laboratory Medicine (M.E., A.E.v.H., F.C.G.J.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands; Centre for Endocrinology, Diabetes and Metabolism, School of Clinical and Experimental Medicine (A.G., N.K.), University of Birmingham, Birmingham B15 2TT, United Kingdom; and Department of Radiation Oncology (P.N.S.), Radboud University Medical Center, 6500HB Nijmegen, The Netherlands.
authors.forename: H L
authors.initials: HL
authors.lastname: Claahsen-van der Grinten
mesh_terms: D019326:17-alpha-Hydroxyprogesterone / Q000737:chemistry; D021581:Active Transport, Cell Nucleus; D000311:Adrenal Glands / Q000378:metabolism*; D000312:Adrenal Hyperplasia, Congenital / Q000378:metabolism*; D000735:Androstenedione / Q000737:chemistry; D000818:Animals; D001667:Binding, Competitive; D019556:COS Cells; D002522:Chlorocebus aethiops; D003350:Cortodoxone / Q000737:chemistry; D005938:Glucocorticoids / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D011374:Progesterone / Q000737:chemistry; D011485:Protein Binding; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D013256:Steroids / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D013256:Steroids; D049452:Green Fluorescent Proteins; D000735:Androstenedione; D011374:Progesterone; C003556:21-deoxycortisol; D019326:17-alpha-Hydroxyprogesterone; D003350:Cortodoxone