Familial glucocorticoid resistance caused by a splice site deletion in the human glucocorticoid receptor gene.

Abstract

The clinical syndrome of generalized, compensated glucocorticoid resistance is characterized by increased cortisol secretion without clinical evidence of hyper- or hypocortisolism, and manifestations of androgen and/or mineralocorticoid excess. This condition results from partial failure of the glucocorticoid receptor (GR) to modulate transcription of its target genes. We studied the molecular mechanisms of this syndrome in a Dutch kindred, whose affected members had hypercortisolism and approximately half of normal GRs, and whose proband was a young woman with manifestations of hyperandrogenism. Using the polymerase chain reaction to amplify and sequence each of the nine exons of the GR gene alpha, along with their 5'- and 3'-flanking regions, we identified a 4-base deletion at the 3'-boundary of exon 6 in one GR allele (delta 4), which removed a donor splice site in all three affected members studied. In contrast, the sequence of exon 6 in the two unaffected siblings was normal. A single nucleotide substitution causing an amino acid substitution in the amino terminal domain of the GR (asparagine to serine, codon 363) was also discovered in exon 2 of the other allele (G1220) in the proband, in one of her affected brothers and in her unaffected sister. The functional importance of this mutation was tested in a cotransfection study using the recombinant expression vector pRShGR-Ser363 and the glucocorticoid responsive vector mouse mammary tumor virus-chloramphenicol transferase. This amino acid substitution did not alter the function of the glucocorticoid receptor. Using reverse transcription-polymerase chain reaction we could only identify messenger RNA transcripts of the G1220-allele but not of the delta 4-allele in the affected members of this family who were heterozygous for the G1220 mutation. This deletion in the glucocorticoid receptor gene was, thus, associated with the expression of only one allele and a decrease of GR protein by 50% in affected members of this glucocorticoid resistant family. The mutation identified in exon 2 did not segregate with the disease and appears to be of no functional significance. The presence of the null allele was apparently compensated for by increased cortisol production at the expense of concurrent hyperandrogenism.

Glucocorticoid receptor alpha and beta isoforms are not mutated in bipolar affective disorder

“…Furthermore, a polymorphic variant of GR, with a change of an Asparagine to Serine at codon 363, has been reported. 31 In the present work we were unable to detect these polymorphisms, possibly due to their low frequency (less than 5%).…”

The role of stress and the hypothalamic–pituitary–adrenal axis in the pathogenesis of the metabolic syndrome: neuro-endocrine and target tissue-related causes

“…The ®nding of the correlation between an unde®ned nonstructural polymorphism of the glucocorticoid receptor gene and a hypersensitive curve suggests that tissue-limited hypersensitivity to glucocorticoids, or its mirror image, glucocorticoid resistance, exist as earlier hypothesized on a theoretical basis. 15 Huizenga et al 34 recently demonstrated that another polymorphism of the glucocorticoid receptor, which we described earlier, 35 was present in 6% of normal Dutch men and associated with a signi®cantly greater cortisol suppression by dexamethasone, a higher BMI and a lower bone mineral density (BMD) in polymorphism carriers than in noncarriers. More recently, Walker et al 36 reported correlation of increased dermal glucocorticoid sensitivity with relative hypertension, insulin resistance and hyperglycemia.…”

NR3C1 polymorphisms in Brazilians of Caucasian, African, and Asian ancestry: glucocorticoid sensitivity and genotype association

“…The p.ER22/23EK (rs6189 -rs6190), and BclI (rs41423247) were evaluated in 264 individuals, including 230 of Caucasian origin, 31 Afro-Brazilians and only 3 Asian descendants. Exons 3 to 9 and intron/exon boundary regions were amplified in 137 to 176 subjects divided in Caucasian, African, and Asian ancestry by PCR using specific primers previously described (23). PCR reactions were followed by automated direct sequencing performed using a commercial kit (ABI Prism ® Big Dye™ Terminator Cycle Sequencing Ready Reaction kit; Applied Biosystems, Foster City, CA, USA).…”