Violeta Anastasovska1, Mirjana Kocova2, Nikolina Zdraveska3, Maja Stojiljkovic4, Anita Skakic4, Kristel Klaassen4, Sonja Pavlovic4. 1. Genetic Laboratory, Department of Endocrinology and Genetics, University Clinic for Pediatrics, Ss. Cyril and Methodius University in Skopje, Faculty of Medicine, Skopje, Republic of North Macedonia. violeta_anastasovska@yahoo.com. 2. Genetic Laboratory, Department of Endocrinology and Genetics, University Clinic for Pediatrics, Ss. Cyril and Methodius University in Skopje, Faculty of Medicine, Skopje, Republic of North Macedonia. 3. Department of Neonatology, University Clinic for Pediatrics, Ss. Cyril and Methodius University in Skopje, Faculty of Medicine, Skopje, Republic of North Macedonia. 4. Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
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.
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.
Authors: V Dolzan; J Sólyom; G Fekete; J Kovács; V Rakosnikova; F Votava; J Lebl; Z Pribilincova; S M Baumgartner-Parzer; S Riedl; F Waldhauser; H Frisch; M Stopar-Obreza; C Krzisnik; T Battelino Journal: Eur J Endocrinol Date: 2005-07 Impact factor: 6.664
Authors: Débora de Paula Michelatto; Leif Karlsson; Ana Letícia Gori Lusa; Camila D'Almeida Mgnani Silva; Linus Joakim Östberg; Bengt Persson; Gil Guerra-Júnior; Sofia Helena Valente de Lemos-Marini; Michela Barbaro; Maricilda Palandi de Mello; Svetlana Lajic Journal: Int J Endocrinol Date: 2016-09-19 Impact factor: 3.257