Differences in binding of glucocorticoid receptor to DNA in steroid-resistant asthma.
Although glucocorticosteroids are a very effective treatment for asthma and other chronic inflammatory diseases, a small proportion of patients are resistant to their therapeutic effects. The molecular mechanism for this steroid resistance is unclear. Steroid resistance cannot be explained by pharmacokinetic mechanisms, by a defect in the binding of steroids to glucocorticoid receptors, nor by defective nuclear translocation of this receptor, thereby suggesting that the molecular abnormality lies distal to nuclear translocation. We examined the ability of nuclear translocated glucocorticoid receptors to bind to their DNA binding sites (GRE) using electrophoretic mobility shift assays in PBMC from patients with steroid-sensitive and steroid-resistant asthma. The binding of the glucocorticoid receptor to DNA in these patients was also studied using Scatchard analysis. Dexamethasone induced a significant rapid and sustained twofold increase in GRE binding in PBMCs from steroid-sensitive asthmatic patients and nonasthmatic individuals, but this was markedly reduced in steroid-resistant asthmatic patients. Scatchard analysis of glucocorticoid receptor-GRE binding showed no change in binding affinity but did show a reduced number of receptors available for DNA binding in the steroid-resistant patients. These results suggest that the ability of the glucocorticoid receptor to bind to GRE is impaired in steroid-resistant patients because of a reduced number of receptors available for binding to DNA.
Adcock
I M
IM
Department of Thoracic Medicine, National Heart and Lung Institute, London, UK.
Lane
S J
SJ
Brown
C R
CR
Peters
M J
MJ
Lee
T H
TH
Barnes
P J
PJ
D000328:Adult; D001249:Asthma / Q000188:drug therapy* / Q000378:metabolism; D001483:Base Sequence; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004351:Drug Resistance / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005938:Glucocorticoids / Q000627:therapeutic use*; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D011965:Receptors, Glucocorticoid / Q000378:metabolism*
D004268:DNA-Binding Proteins; D005938:Glucocorticoids; D016209:Interleukin-8; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid