Exp Clin Endocrinol Diabetes 2010; 118(8): 505-512
DOI: 10.1055/s-0029-1237363
Article

© J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York

Mutational Characterization of Steroid 21-Hydroxylase Gene in Portuguese patients with Congenital Adrenal Hyperplasia

C. J. Marques1 , D. Pignatelli2 , B. Carvalho1 , J. Barceló1 , A. C. Almeida1 , S. Fernandes1 , S. F. Witchel3 , M. Sousa4 , M. J. Oliveira5 , P. Freitas2 , M. Fontoura6 , D. Carvalho2 , A. Barros1 , F. Carvalho1
  • 1Department of Genetics, Faculty of Medicine, University of Porto, Porto, Portugal
  • 2Department of Endocrinology, S. João Hospital, Porto, Portugal
  • 3Division of Pediatric Endocrinology, Children's Hospital of Pittsburgh, University of Pittsburgh, Pittsburgh, PA, USA
  • 4Lab of Cell Biology, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Portugal
  • 5Department of Endocrinology, Pedro Hispano Hospital, Matosinhos, Portugal
  • 6Department of Pediatrics, S. João Hospital, Porto, Portugal
Further Information

Publication History

received 07.04.2009 first decision 02.06.2009

accepted 26.06.2009

Publication Date:
23 October 2009 (online)

Abstract

Congenital adrenal hyperplasia (CAH) due to steroid 21-hydroxylase deficiency is a common inherited disorder of adrenal hormone biosynthesis due to mutations in the 21-hydroxylase gene, CYP21A2. Genotyping for ten of the most frequent mutations was performed in 84 Portuguese CAH patients: 10 salt-wasters, 6 simple-virilizers and 68 non-classical patients. The patients were diagnosed by a level of 17-hydroxyprogesterone above 10 ng/ml either in basal conditions or after an ACTH 0,25 mg IV Test. A variety of genotyping techniques were used to detect these ten mutations. CYP21A2 mutations were detected in 91.7% (77/84) of the patients. The frequency of alleles carrying two or more CYP21A2 mutations (9.5% – 16/168) is higher than in other populations. The most frequent mutations identified in our population were V281L (41.7%) and deletions/conversions involving the promoter region of the CYP21A2 gene (28.3%). A decreased frequency of IVS2-12C/A>G mutation (5.6%) was the most characteristic feature of our population. This study allow the characterization of the mutational spectrum of CAH patients, mainly non-classical CAH, with 21-hydroxylase deficiency from Portugal showing specific genetic features of this population which reveals differences with worldwide countries.

References

  • 1 Asanuma A, Ohura T, Ogawa E. et al . Molecular analysis of Japanese patients with steroid 21-hydroxylase deficiency.  J Hum Genet. 1999;  44 312-317
  • 2 Bleicken C, Loidi L, Dhir V. et al . Functional characterization of three CYP21A2 sequence variants (p.A265V, p.W302S, p.D322G) employing a yeast co-expression system.  Hum Mutat. 2008;  29 E43-E450
  • 3 Carrera P, Bordone L, Azzani T. et al . Point mutations in Italian patients with classic, non-classic, and cryptic forms of steroid 21-hydroxylase deficiency.  Hum Genet. 1996;  98 662-665
  • 4 Carroll MC, Campbell RD, Porter RR. Mapping of steroid 21-hydroxylase genes adjacent to complement component C4 genes in HLA, the major histocompatibility complex in man.  Proc Natl Acad Sci USA. 1985;  82 521-525
  • 5 Chang S, Chung B. Difference in transcriptional activity of two homologous CYP21A genes.  Mol Endocrinol. 1995;  9 1330-1336
  • 6 Deneux C, Tardy V, Dib A. et al . Phenotype-genotypecorrelation in 56 women with nonclassical congenital adrenal hyperplasia due to 21-hydroxylasedeficiency.  J Clin Endocrinol Metab. 2001;  86 ((1)) 207-213
  • 7 Dolzan V, Sólyom J, Fekete G. et al . Mutational spectrum of steroid 21-hydroxylase and the genotype-phenotype association in MiddleEuropean patients with congenital adrenal hyperlasia.  Eur J Endocrinol. 2005;  153 99-106
  • 8 Ezquieta B, Cueva E, Oyarzábal M. et al . Gene conversion (655G splicing mutation) and the founder effect (Gln318Stop) contribute to the most frequent severe point mutations in congenital adrenal hyperplasia (21-hydroxylase deficiency) in the Spanish population.  Clin Genet. 2002;  62 181-188
  • 9 Ezquieta B, Oliver A, Gracia R. et al . Analysis of steroid 21-hydroxylase gene mutations in theSpanish population.  Hum Genet. 1995;  96 198-204
  • 10 Friães A, Rêgo AT, Aragüés JM. et al . CYP21A2 mutations in Portuguese patients with congenital adrenal hyperplasia: identification of two novel mutations and characterization of four different partial gene convertions.  Mol Genet Metab. 2006;  88 58-65
  • 11 Higashi Y, Tanae A, Inoue H. et al . Aberrant splicing and missensemutations cause steroid 21-hydroxylase [P-450(C21)] deficiency in humans: Possible gene conversion products.  Proc Natl Acad Sci USA. 1998;  85 7486-7490
  • 12 Kostaska K, Lisá L, Prusa R. Common CYP21 gene mutations in Czech patients and statistical analysisof worldwide mutation distribution.  Cent Eur J Public Health. 2003;  11 124-128
  • 13 Krone N, Braun A, Roscher AA. et al . Predicting phenotype in steroid 21-hydroxylase deficiency? Comprehensive genotyping in 155 unrelated, well defined patients from southern germany.  J Clin Endocrinol Metab. 2000;  85 ((3)) 1059-1065
  • 14 Lako M, Ramsden S, Campbell RD. et al . Mutation screening in British 21-hydroxylasedeficiency families and development of novel microsatellite based approaches to prenatal diagnosis.  J Med Genet. 1999;  36 119-124
  • 15 Lee HH, Chang JG, Tsai CH. et al . Analysis of the Chimeric CYP21P/CYP21 Gene in Steroid 21-Hydroxylase Deficiency.  Clin Chemistry. 2000;  56 ((5)) 606-611
  • 16 Levo A, Partanen J. Mutation-haplotype analysis of steroid 21-hydroxylase (CYP21) deficiency in Finland. Implications for the population history of defective alleles.  Hum Genet. 1997;  99 488-497
  • 17 Merke DP, Bornstein SR. Congenital adrenal hyperplasia.  Lancet. 2005;  365 2125-2136
  • 18 Mullenbach R, Lagoda PJ, Welter C. An efficient salt-chloroform extraction of DNA from blood and tissues.  Trends Genet. 1989;  5 ((12)) 391
  • 19 Nikoshkov A, Lajic S, Holst M. et al . Synergetic effect of partially inactivating mutations in steroid 21-hydroxylase deficiency.  J Clin Endocrinol Metab. 1997;  82 ((1)) 194-199
  • 20 Ordoñez-Sánchez ML, Ramírez-Jiménez S, López-Gutierrez AU. et al . Molecular genetic analysis of patients carrying steroid 21-hydroxylase deficiency inthe Mexican population: identification of possible new mutations and high prevalence of apparent germ-line mutations.  Hum Genet. 1998;  102 170-177
  • 21 Pang SY, Wallace MA, Hofman L. et al . Worldwide experience in newborn screening for classical congenital adrenal hyperplasia due to 21-hydroxylase deficiency.  Pediatrics. 1988;  81 866-874
  • 22 Rodrigues NR, Dunham I, Yu CY. et al . Molecular characterizationof the HLA-linked steroid 21-hydroxylase B gene from an individual with congenital adrenal hyperplasia.  The EMBO J. 1987;  6 ((6)) 1653-1661
  • 23 Rothberg PG, Baker DW, Bradley JF. Simultaneous Detection of Five Mutations in the Steroid 21-Hydroxylase Gene Using Nested Allele-Specific Amplification.  Genetic Testing. 1998;  2 ((4)) 343-346
  • 24 Siegel SF, Hoffman EP, Trucco M. Molecular diagnosis of 21-Hidroxylase deficiency: detection of four mutations on a single gel.  Biochem Med Metab Biol. 1994;  51 66-73
  • 25 Siegel SF, Lee PA, Rudert WA. et al . Phenotype/genotype correlations in 21-hydroxylase deficiency.  Adol Ped Gynecol. 1995;  8 9-16
  • 26 Siegel-Witchel S, Lee PA, Trucco M. Who is a carrier? Detection of unsuspected mutations in 21 hydroxylase deficiency.  Am J Med Genet. 1996;  61 2-9
  • 27 Speiser PW, Dupont J, Zhu D. et al . Disease expression and molecular genotype in congenital adrenal hyperplasia due to 21-hydroxylase deficiency.  J Clin Invest. 1992;  90 584-595
  • 28 Speiser PW, Knochenhauer ES, Dewailly D. et al . A multicenter study of women with nonclassical congenital adrenal hyperplasia: relationship between genotype and phenotype.  Mol Genet Metab. 2000;  71 527-534
  • 29 Speiser PW, White PC. Congenital adrenal hyperplasia.  N Engl J Med. 2003;  349 776-788
  • 30 Stikkelbroeck NMML, Hoefsloot LH, Wijs IJ. et al . CYP21 gene mutation analysis in 198 patients with 21-hydroxylase deficiency in the Netherlands: six novel mutations and a specific cluster of four mutations.  J Clin Endocrinol Metab. 2003;  88 ((8)) 3852-3859
  • 31 Tusie-Luna MT, White PC. Gene conversions and unequal crossover between CYP21 (steroid 21-hydroxylase gene) and CYP21A1P involve different mechanisms.  Proc Natl Acad Sci USA. 1995;  92 10796-10800
  • 32 Wedell A, Ritzén EM, Haglund-Stengler B. et al . Steroid 21-hydroxylase deficiency: three additional mutated alleles and establishment of phenotype-genotype relationships of common mutations.  Proc Natl Acad Sci USA. 1992;  89 7232-7236
  • 33 Wedell A, Thilen A, Ritzen M. et al . Mutational spectrum of the steroid 21 hydroxylase gene in Sweden: implications for genetic diagnosis and association with disease manifestation.  J Clin Endocrinol Metab. 1994;  8 ((5)) 1145-1152
  • 34 White PC, Grossberger D, Onufer BJ. et al . Two genes encoding steroid 21-hydroxylase are located near the genes encoding the fourth component of complement in man.  Proc Natl Acad Sci USA. 1985;  82 1089-1093
  • 35 White PC, New MI, Dupont B. Structure of human steroid 21-hydroxylase genes.  Proc Natl Acad Sci USA. 1986;  83 5111-5115
  • 36 White PC, Speiser PW. Congenital Adrenal Hyperplasia due to 21-Hydroxylase Deficiency.  Endocrine Reviews. 2000;  21 ((3)) 245-291
  • 37 White PC, Vitek A, Dupont B. et al . Characterization of frequent deletions causing steroid 21 hydroxylase deficiency.  Proc Natl Acad Sci USA. 1988;  85 4436-4440
  • 38 Wilson RC, Mercado AB, Cheng KC. et al . Steroid 21-hydroxylase deficiency: genotype may notpredict phenotype.  J Clin Endocrinol Metab. 1995;  80 ((8)) 2322-2329
  • 39 Wilson RC, Nimkarn S, Dumic M. et al . Ethnic-specific distribution of mutations in 716 patients with congenital adrenal hyperplasia owing to 21-hydroxylase deficiency.  Mol Genet Metab. 2007;  90 414-421
  • 40 Witchel SF, Nayak S, Suda-Hartman M. et al . Newborn screening for 21-hydroxylase deficiency: results of CYP21A2 molecular analysis.  J Pediat. 1997;  131 328-331
  • 41 Witchel SF, Smith R, Crivellaro CE. et al . CYP21 mutations in Brazilian patients with 21-hydroxylase deficiency.  Hum Genet. 2000;  106 414-419
  • 42 Wu DA, Chung BJ. Mutations of P450c21 (steroid 21-Hydroxylase) at Cys428, Val281, and Ser268 result in complete, partial, or no loss of enzymatic activity, respectively.  J Clin Invest. 1991;  88 519-523

Correspondence

F. CarvalhoPhD 

Department of Genetics

Faculty of Medicine

University of Porto

Alameda Prof Hernâni Monteiro

4200-319 Porto

Portugal

Phone: +351 22 551 36 47

Fax: +351 22 551 36 48

Email: filipac@med.up.pt