A sequence variation in 3’UTR of CYP21A2 gene correlates with a mild form of congenital adrenal hyperplasia.

Background Congenital adrenal hyperplasia is mainly caused by the deficiency of the 21-hydroxylase enzyme coded by the CYP21A2 gene. However, some alleles in the nonclassical form (NC-CAH) remain without identified mutations, suggesting the involvement of regulatory regions. Aim Our objective was to study an allele carrying the variant *13 G>A in the 3’UTR of the CYP21A2 gene identified in some patients with a mild form of NC-CAH in order to verify the possible implication of this variation with the phenotype observed. Subjects and Methods Among all the subjects in whom the CYP21A2 gene was analysed, 14 patients and 7 relatives heterozygous or homozygous for the *13 G>A substitution in 3’UTR were selected. Sequencing of DNA, genotyping, multiplex ligation-dependent probe amplification (MLPA), in-vitro studies and bioinformatic analysis were performed. Results The haplotype of the *13 G>A allele was identical in all the subjects with a monomodular structure composed by one C4A gene and one CYP21A2 gene without a second module with the CYP21A1P pseudogene. No other concomitant mutations were found in the region extending from 3 Kb in the promoter and encompassing the polyadenylation signal. Both bioinformatic analysis and in vitro studies predicted an alteration of the RNA folding and expression, but no miRNA target sequences were found in this region. Conclusions The identification of a substitution in the 3’UTR of the gene associated with a mild form of NC-CAH suggests the importance of analysing the CYP21A2 untranslated regions to better characterize and treat this subgroup of patients.