Genetic diagnosis of 21-hydroxylase deficiency: DGGE-based mutation scanning of CYP21

Ohlsson, Gita; Müller, Jørn; Schwartz, Marianne

doi:10.1002/(sici)1098-1004(1999)13:5<385::aid-humu7>3.0.co;2-2

Cited by 8 publications

(7 citation statements)

References 28 publications

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“…Genotyping at positions for known disease‐causing mutations was performed by direct DNA sequencing and allele‐specific polymerase chain reaction (PCR) on DNA isolated from peripheral blood lymphocytes (Wedell et al. , 1994; Ohlsson et al. , 1999).…”

Section: Methodsmentioning

confidence: 99%

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Testicular adrenal rest tumours in boys, adolescents and adult men with congenital adrenal hyperplasia may be associated with the CYP21A2 mutation

Mouritsen¹,

Jørgensen²,

Main³

et al. 2010

Int J of Andrology

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Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder with impaired function of the adrenal cortex caused by mutations in the CYP21A2 gene. Deficiency of steroid 21-hydroxylase accounts for 80-95% of CAH cases. Testicular adrenal rest tumours (TART) may be prevalent in up to 95% of CAH adults and may already appear during childhood. Whether genotype sub-types can account for the development of TART has not been investigated previously. We therefore investigated this by coupling clinical information of CAH patients with information of their genetic mutation. In 49 male patients (age 2.6-40.3 years) with 21-hydroxylase deficiency, testicular ultrasound examinations were performed and CYP21A2 genotypes determined. These were grouped according to the residual 21-hydroxylase activity: group Null (complete enzyme impairment), group A (almost complete enzyme impairment), group B (severe enzyme impairment) and group C (partial impairment). TART were observed in 27 of 49 patients (55%). For the 23 patients younger than 18 years, TART were present in 11 (48%), the youngest patient being 7.5 years old. The presence of TART was dependent on the CYP21A2 genotype: 27 of 37 patients (73%) with the most severe mutations (groups Null and A) had TART, whereas none of 12 patients with the milder mutations (groups B and C) had TART. We conclude that TART were most frequently detected in patients with severe CYP21A2 mutations, and may occur already in early childhood in such patients.

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Section: Methodsmentioning

confidence: 99%

“…, 2001). Steroid 21‐hydroxylase deficiency accounts for 80–95% of CAH cases (Ohlsson et al. , 1999), and the specific mutation determines the degree of impairment of 21‐hydroxylase activity (Wedell et al.…”

Section: Introductionmentioning

confidence: 99%

Testicular adrenal rest tumours in boys, adolescents and adult men with congenital adrenal hyperplasia may be associated with the CYP21A2 mutation

Mouritsen¹,

Jørgensen²,

Main³

et al. 2010

Int J of Andrology

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“…CAH is predominantly (80-95% of cases) caused by mutations in the CYP21A2 gene encoding the enzyme steroid 21-hydroxylase (cytochrome P-450c21) (1,2).…”

Section: Introductionmentioning

confidence: 99%

Abundance of DLK1, differential expression of CYP11B1, CYP21A2 and MC2R, and lack of INSL3 distinguish testicular adrenal rest tumours from Leydig cell tumours

Lottrup

Nielsen

Skakkebæk

et al. 2015

European Journal of Endocrinology

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Objective: Testicular adrenal rest tumours (TARTs) are a common finding in patients with congenital adrenal hyperplasia (CAH). These tumours constitute a diagnostic and management conundrum and may lead to infertility. TART cells share many functional and morphological similarities with Leydig cells (LCs), and masses consisting of such cells are occasionally misclassified as malignant testicular tumours, which may lead to erroneous orchiectomy in these patients. Design: In this study, we aimed to investigate the potential of LC developmental markers and adrenal steroidogenic markers in the differential diagnosis of TARTs and malignant LC tumours (LCTs). Methods: We investigated mRNA and protein expression of testicular steroidogenic enzymes; CYP11A1 and HSD3B1/2, markers of adrenal steroidogenesis; CYP11B1, CYP21A2 and ACTH receptor/melanocortin 2 receptor (MC2R), and markers of LC maturation; and delta-like 1 homolog (DLK1) and insulin-like 3 (INSL3) in testicular biopsies with TART, orchiectomy specimens with LCTs and samples from human fetal adrenals. Results: Expression of testicular steroidogenic enzymes was observed in all specimens. All investigated adrenal steroidogenic markers were expressed in TART, and weak reactions for CYP11B1 and MC2R were observed at the protein level in LTCs. TART and fetal adrenals had identical expression profiles. DLK1 was highly expressed and INSL3 not detectable in TART, whereas INSL3 was highly expressed in LCTs. Conclusions: The similar expression profiles in TART and fetal adrenals as well as the presence of classical markers of adrenal steroidogenesis lend support to the hypothesis that TART develops from a displaced adrenal cell type. Malignant LCTs seem to have lost DLK1 expression and do not resemble immature LCs. The different expression pattern of DLK1, INSL3 and most adrenocortical markers adds to the elucidation of the histogenesis of testicular interstitial tumours and may facilitate histopathological diagnosis.

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“…These mutations can be identified by allele-specific oligonucleotide (ASO) (Speiser et al 1994) and reverse dot-blot hybridizations (Yang et al 2001), direct DNA sequencing (Tajima et al 1993), PCR/ligase (Day et al 1995), and the amplification-created restriction site (ACRS) method (Lee et al 1996). For some unknown defective loci somewhere within the CYP21 gene, SSCP (Tajima et al 1993;Lee et al 1998) and denatured gradient gel electrophoresis (DGGE) (Ohlsson et al 1999) may be applied to possibly find novel mutations in the CYP21 gene. However, all these applications for mutational detection should be carried out using the functional CYP21 gene.…”

Section: Introductionmentioning

confidence: 99%

The chimeric CYP21P/CYP21 gene and 21-hydroxylase deficiency

Lee¹

2004

J Hum Genet

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The chimeric CYP21P/CYP21 gene is a consequence of a 26-or 32-kb deletion in the C4-CYP21 repeat module of CYP21P, tenascin A (XA), serine/ threonine nuclear protein kinase (RP2), and the C4B and CYP21 genes in congenital adrenal hyperplasia (CAH) with steroid 21-hydroxylase deficiency. To date, there have been three distinct chimeras found in CAH patients in ethnic Chinese. Initiation for production of these molecules is proposed to be chi-like sequences and a minisatellite consensus existing in several noncoding regions in CYP21 genes. These molecules have the 5' end of the CYP21P-specific sequence in common but differ in the 3' end of CYP21-specific genes. In addition, there appears to be a 3.2-kb fragment generated by Taq I digestion, which leads to allele dropout in PCR amplification for detecting the aberrant splicing site of the IVS2 )12A/C>G mutation at nucleotide (nt) 655 in the CYP21 gene. Therefore, the chimeric CYP21P/CYP21 cannot be detected by conventional methods. It has been demonstrated that a PCR product amplified with allele-specific primers covering tenascin B (TNXB) to the 5' end of the CYP21 gene combined with Southern analysis by Ase I and Nde I digestion may be used for identifying the chimera in the CYP21 gene.

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Genetic diagnosis of 21-hydroxylase deficiency: DGGE-based mutation scanning of CYP21

Cited by 8 publications

References 28 publications

Testicular adrenal rest tumours in boys, adolescents and adult men with congenital adrenal hyperplasia may be associated with the CYP21A2 mutation

Testicular adrenal rest tumours in boys, adolescents and adult men with congenital adrenal hyperplasia may be associated with the CYP21A2 mutation

Abundance of DLK1, differential expression of CYP11B1, CYP21A2 and MC2R, and lack of INSL3 distinguish testicular adrenal rest tumours from Leydig cell tumours

The chimeric CYP21P/CYP21 gene and 21-hydroxylase deficiency

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