Two novel CYP21A2 missense mutations in Italian patients with 21‐hydroxylase deficiency: Identification and functional characterisation

Steroid 21‐hydroxylase deficiency is present in more than 90% of patients with congenital adrenal hyperplasia (CAH), an inherited metabolic disorder of adrenal steroidogenesis. Impaired enzymatic activity leads to the accumulation of metabolic intermediates (progesterone and 17‐hydroxyprogesterone), which results in excessive androgen production and varied signs of virilisation. CYP 21A2 is an active gene and encodes for the steroid 21‐hydroxylase enzyme, whereas CYP21A1P is an inactive pseudogene that contains a series of deleterious mutations. The major part of disease‐ causing mutations in CYP21A2 alleles are CYP21A1P ‐derived sequence transferred to the active gene by macro or microconversion events. Only around 5% of all disease‐causing CYP21A2 alleles harbour rare mutations that do not originate from the pseudogene. In this report, we report the characterisation of two novel CYP21A2 missense mutations (p.H119R and p.I194N) found in two unrelated Italian patients with nonclassic (NC) CAH clinical diagnosis. Functional in vitro assays for mutagenized CYP21 enzymes were performed in transiently transfected mammalian cells to test the residual enzyme activity and the apparent kinetic values. The residual activities obtained for p.H119R and p.I194N mutants allowed to classify them as NC‐CAH associated mutations. These results correlate with the rate of severity of the patients' disease. Finally, the new p.H119R and p.I194N mutations should be included in the panel of those already listed for association with the NC form of 21‐hydroxylase deficiency. © 2009 IUBMB IUBMB Life 61(3): 229–235, 2009