This Article Full Text Full Text (PDF) Submit a related Letter to the Editor Purchase Article View Shopping Cart Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Vermeer, H. Articles by Jansen, M. Search for Related Content PubMed PubMed Citation Articles by Vermeer, H. Articles by Jansen, M.

Glucocorticoid-Induced Increase in Lymphocytic FKBP51 Messenger Ribonucleic Acid Expression: A Potential Marker for Glucocorticoid Sensitivity, Potency, and Bioavailability

Department of Pediatric Endocrinology (H.V., B.I.H.-S., S.C.V.B.-O., M.J.), University Medical Center Utrecht, 3508 AB Utrecht; and Netherlands Institute of Developmental Biology (B.v.d.B.), 3584 CT Utrecht, The Netherlands

Address all correspondence and requests for reprints to: M. Jansen, Department of Pediatric Endocrinology, HP KC.03.063.0, Wilhelmina Children’s Hospital, University Medical Center Utrecht, P.O. Box 85090, 3508 AB, Utrecht, The Netherlands. E-mail: m.jansen{at}wkz.azu.nl.

To reduce the side effects of corticosteroid treatment, the administered dose of glucocorticoids (GCs) should be kept to a minimum while preserving therapeutically needed intracellular levels. Currently available assays to determine individual sensitivity to GCs are either imprecise or based on inhibition by GCs of lymphocyte proliferation following stimulation with phytohemagglutinin or other mitogens, which may influence the GC signal transduction pathway. Using the human lymphoblastoid cell line IM-9 as a model system, we studied whether the GC-induced increase of the mRNA encoding the 51-kDa FK506-binding protein (FKBP51) could be used for the development of a novel assay, ultimately to be used in native human peripheral blood mononuclear cells (PBMCs). GC addition to IM-9 cells resulted in a dose-dependent increase of FKBP51 mRNA levels within 2 h, followed by a further increase until 24 h. Northern blot analysis and real-time PCR yielded similar results. Coincubation of GCs with the GC receptor antagonist ORG 34116 or the protein synthesis inhibitor cycloheximide suggested a direct, GC receptor-mediated up-regulation of FKBP51 gene transcription. Expected differences in potency among different GCs could be readily demonstrated in this system. Extending our observations in IM-9 lymphoblasts to normal PBMCs, we found a dose-dependent increase of FKBP51 mRNA on ex vivo incubation of native human PBMCs with GCs, with a sensitivity of about 10^-9 M for dexamethasone. Moreover, dexamethasone ingestion increased FKBP51 mRNA in PBMCs in vivo, extending the use of this assay to the measurement of GC bioavailability. Finally, using this method we were able to demonstrate partial GC-insensitivity in a 6-month-old infant suffering from congenital adrenal hyperplasia caused by 21-hydroxylase deficiency. We conclude that the induction of FKBP51 mRNA by GCs may be a suitable marker to assess individual GC sensitivity, the in vitro measurement of GC potency, and the in vivo determination of GC bioavailability.

This work was supported by a research grant from Ferring Pharmaceuticals Ltd. Netherlands, The Netherlands (to H.V.).

Abbreviations: ß₂m, ß₂-Microglobulin; dsDNA, double-strand DNA; FCS, fetal calf serum; FKBP51, 51-kDa FK506-binding protein; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GC, glucocorticoid; GRE, glucocorticoid response element.

This article has been cited by other articles:

				S. R. Holmstrom, S. Chupreta, A. Y.-L. So, and J. A. Iniguez-Lluhi SUMO-Mediated Inhibition of Glucocorticoid Receptor Synergistic Activity Depends on Stable Assembly at the Promoter But Not on DAXX Mol. Endocrinol., September 1, 2008; 22(9): 2061 - 2075. [Abstract] [Full Text] [PDF]

				M. C. Haffner, A. Jurgeit, C. Berlato, S. Geley, N. Parajuli, A. Yoshimura, and W. Doppler Interaction and Functional Interference of Glucocorticoid Receptor and SOCS1 J. Biol. Chem., August 8, 2008; 283(32): 22089 - 22096. [Abstract] [Full Text] [PDF]

				E. B. Binder, R. G. Bradley, W. Liu, M. P. Epstein, T. C. Deveau, K. B. Mercer, Y. Tang, C. F. Gillespie, C. M. Heim, C. B. Nemeroff, et al. Association of FKBP5 Polymorphisms and Childhood Abuse With Risk of Posttraumatic Stress Disorder Symptoms in Adults JAMA, March 19, 2008; 299(11): 1291 - 1305. [Abstract] [Full Text] [PDF]

				P. G. Woodruff, H. A. Boushey, G. M. Dolganov, C. S. Barker, Y. H. Yang, S. Donnelly, A. Ellwanger, S. S. Sidhu, T. P. Dao-Pick, C. Pantoja, et al. From the Cover: Genome-wide profiling identifies epithelial cell genes associated with asthma and with treatment response to corticosteroids PNAS, October 2, 2007; 104(40): 15858 - 15863. [Abstract] [Full Text] [PDF]

				W. J. E. Tissing, M. L. den Boer, J. P. P. Meijerink, R. X. Menezes, S. Swagemakers, P. J. van der Spek, S. E. Sallan, S. A. Armstrong, and R. Pieters Genomewide identification of prednisolone-responsive genes in acute lymphoblastic leukemia cells Blood, May 1, 2007; 109(9): 3929 - 3935. [Abstract] [Full Text] [PDF]

				T. Kino, E. Souvatzoglou, E. Charmandari, T. Ichijo, P. Driggers, C. Mayers, A. Alatsatianos, I. Manoli, H. Westphal, G. P. Chrousos, et al. Rho Family Guanine Nucleotide Exchange Factor Brx Couples Extracellular Signals to the Glucocorticoid Signaling System J. Biol. Chem., April 7, 2006; 281(14): 9118 - 9126. [Abstract] [Full Text] [PDF]

				S. Schmidt, J. Rainer, S. Riml, C. Ploner, S. Jesacher, C. Achmuller, E. Presul, S. Skvortsov, R. Crazzolara, M. Fiegl, et al. Identification of glucocorticoid-response genes in children with acute lymphoblastic leukemia Blood, March 1, 2006; 107(5): 2061 - 2069. [Abstract] [Full Text] [PDF]

				G. G.-v. Simson, B. Kohn, and F. B. Axelrod Cushing Syndrome from Topical Foam Steroid Use in an Adolescent Male Clinical Pediatrics, January 1, 2006; 45(1): 97 - 100. [PDF]

				P. Smit, H. Russcher, F. H. de Jong, A. O. Brinkmann, S. W. J. Lamberts, and J. W. Koper Differential Regulation of Synthetic Glucocorticoids on Gene Expression Levels of Glucocorticoid-Induced Leucine Zipper and Interleukin-2 J. Clin. Endocrinol. Metab., May 1, 2005; 90(5): 2994 - 3000. [Abstract] [Full Text] [PDF]

				J.-C. Wang, M. K. Derynck, D. F. Nonaka, D. B. Khodabakhsh, C. Haqq, and K. R. Yamamoto From The Cover: Chromatin immunoprecipitation (ChIP) scanning identifies primary glucocorticoid receptor target genes PNAS, November 2, 2004; 101(44): 15603 - 15608. [Abstract] [Full Text] [PDF]

				K. Oishi, K. Miyazaki, K. Kadota, R. Kikuno, T. Nagase, G.-i. Atsumi, N. Ohkura, T. Azama, M. Mesaki, S. Yukimasa, et al. Genome-wide Expression Analysis of Mouse Liver Reveals CLOCK-regulated Circadian Output Genes J. Biol. Chem., October 17, 2003; 278(42): 41519 - 41527. [Abstract] [Full Text] [PDF]

				M. A. de Paula Brotto Temporal Effects of Stress by Immobilization and Sensitivity of the Isolated Rat Pacemaker to Isoproterenol: Roles of Corticosterone, Neuronal Uptake, and {beta}-Adrenergic Homogeneity J. Pharmacol. Exp. Ther., September 1, 2003; 306(3): 1152 - 1158. [Abstract] [Full Text] [PDF]