All forms of 21-hydroxylase deficiency, salt-losing, simple virilizing, and late-onset, share a common gene locus (CYP21B) in the class III region of the major histocompatibility complex on chromosome 6 in tandem duplication with a pseudogene (CYP21A). Gene deletions, conversions, and point mutations have been described in patients with 21-hydroxylase deficiency. This heterogeneity of clinical phenotypes and mutational events suggests that there is a wide variation in the expression of 21-hydroxylase activity. To evaluate our hypothesis that greater disturbance of gene expression leads to greater decrease in 21-hydroxylase activity manifested as more severe phenotypic disease, we propose to correlate ,the frequency of alterations in the structural portion of the 21-hydroxylase gene, detected through genomic DNA amplification utilizing polymerase chain reaction and dot-blot analysis, with the clinical phenotype, clinical severity and 17-hydroxyprogesterone response to synthetic ACTH, in our population of affected families Since defects may occur in the regulatory portion of the gene, we will examine the regulatory portion if there are no apparent nucleotide sequence alterations in the structural portion of the gene. To confirm that detected sequence alterations affect enzyme activity, expression of the altered sequence in pKCRH-2 transfected into COS-7 monkey kidney cells with determination of conversion of 17-hydroxy[14C]progesterone into 14C-lldeoxycortisol will be performed. Results of expressed enzyme activity will be correlated with phenotype and genotype to determine if phenotype predicts genotype.

National Institute of Health (NIH)
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Physician Scientist Award (K11)
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Maternal and Child Health Research Committee (HDMC)
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Children's Hosp Pittsburgh/Upmc Health Sys
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Witchel, S F; Arslanian, S; Lee, P A (1999) Leptin concentrations in precocious puberty or untimely puberty with and without GnRH analogue therapy. J Pediatr Endocrinol Metab 12:839-45
Witchel, S F; Lee, P A (1998) Human chorionic gonadotropin stimulation to assess for ovarian hyperandrogenism. J Pediatr Adolesc Gynecol 11:73-8
Witchel, S F; Lee, P A (1998) Identification of heterozygotic carriers of 21-hydroxylase deficiency: sensitivity of ACTH stimulation tests. Am J Med Genet 76:337-42
Witchel, S F; Lee, P A; Suda-Hartman, M et al. (1997) Hyperandrogenism and manifesting heterozygotes for 21-hydroxylase deficiency. Biochem Mol Med 62:151-8
Witchel, S F; Wenger, S L; Hoffman, E P (1997) Molecular and cytogenetic studies of X inactivation in a patient with 46,X,del(X)(q22). J Pediatr Adolesc Gynecol 10:78-82
Witchel, S F; Nayak, S; Suda-Hartman, M et al. (1997) Newborn screening for 21-hydroxylase deficiency: results of CYP21 molecular genetic analysis. J Pediatr 131:328-31
Witchel, S F; Lee, P A; Suda-Hartman, M et al. (1997) Evidence for a heterozygote advantage in congenital adrenal hyperplasia due to 21-hydroxylase deficiency. J Clin Endocrinol Metab 82:2097-101
Witchel, S F; Bhamidipati, D K; Hoffman, E P et al. (1996) Phenotypic heterogeneity associated with the splicing mutation in congenital adrenal hyperplasia due to 21-hydroxylase deficiency. J Clin Endocrinol Metab 81:4081-8
Witchel, S F; Baens-Bailon, R G; Lee, P A (1996) Treatment of central precocious puberty: comparison of urinary gonadotropin excretion and gonadotropin-releasing hormone (GnRH) stimulation tests in monitoring GnRH analog therapy. J Clin Endocrinol Metab 81:1353-6
Witchel, S S; Lee, P A; Trucco, M (1996) Who is a carrier? Detection of unsuspected mutations in 21-hydroxylase deficiency. Am J Med Genet 61:2-9

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