Abstract
Background
Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder of adrenal steroidogenesis with a broad spectrum of clinical presentations, ranging from the severe classical salt-wasting (SW) and simple-virilizing (SV) form, to the mild nonclassical form. A large variety of CYP21A2 genotypes in correlation with phenotype have been described.
Materials and methods
DNA samples from a 14-day-old male newborn with clinical and laboratory signs of SW CAH and family members were subjected for molecular analysis of the nine most common point CYP21A2 mutations by ACRS/PCR method. Direct DNA sequencing of the whole CYP21A2 gene was performed to detect the second mutant allele in the patient. The in silico predicting analysis and the crystal structure analysis of the mutated CYP21A2 protein have been performed.
Results
Molecular analysis confirmed that the patient was compound heterozygote carrying p.Q318X mutation inherited from the mother and a novel c.1271_1279delGTGCCCGCG (p.G424_R426del) variant in exon 10 inherited from the father. The in silico predicting software tools classified the novel mutation as pathogenic. Crystal structure analysis showed that the three residues affected by the novel in-frame deletion form several hydrogen bonds that could lead to impaired stability and function of the CYP21A2 protein. These findings were concordant with the patient’s phenotype. The need of several molecular methods to elucidate the genotype in this patient has also been discussed.
Conclusions
A novel 9 bp deletion in CYP21A2 gene with predicted pathogenic effect on the enzyme activity was detected in neonatal patient causing severe SW CAH.
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Data availability
The data sets used and/or analyzed during the current study are available from the corresponding author.
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We thank the patient for getting permission to publish relative information.
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All authors contributed to the study conception and design. The clinical diagnosis, treatment, and follow-up of the patient were performed by M.K. and N.Z. who conceived the paper. Material preparation, and ACRS/PCR molecular analysis as well as the discussion of molecular findings were performed by V.A. Sanger sequencing analysis was performed by M.S., A.S., and S.P. The model of the CYP21A2 protein using the crystal structure was made by K.K. All authors read and approved the final paper.
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This study was approved by the Human Research Ethics Committee of University Pediatric Clinic, Medical Faculty, Skopje, North Macedonia, and conducted under the instructions of the principles in the Declaration of Helsinki.
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Anastasovska, V., Kocova, M., Zdraveska, N. et al. A novel 9 bp deletion (c.1271_1279delGTGCCCGCG) in exon 10 of CYP21A2 gene causing severe congenital adrenal hyperplasia. Endocrine 73, 196–202 (2021). https://doi.org/10.1007/s12020-021-02680-7
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DOI: https://doi.org/10.1007/s12020-021-02680-7