The 21-hydroxylase deficiency (21OHD) is caused by CYP21A2 mutations resulting in severe or moderate enzymatic impairments. 21OHD females carrying similar genotypes present different degrees of external genitalia virilization, suggesting the influence of other genetic factors. Single nucleotide variants (SNVs) in the CYP3A7 gene and in its transcription factors, related to fetal 19-carbon steroid metabolism, could modulate the genital phenotype. To evaluate the influence of the 21OHD genotypes and the CYP3A7, PXR and CAR SNVs on the genital phenotype in 21OHD females. Prader scores were evaluated in 183 patients. The CYP3A7, PXR and CAR SNVs were screened and the 21OHD genotypes were classified according to their severity: severe and moderate groups. Patients with severe genotype showed higher degree of genital virilization (Prader median III, IQR III-IV) than those with moderate genotype (III, IQR II-III) (p < 0.001). However, a great overlap was observed between genotype groups. Among all the SNVs tested, only the CAR rs2307424 variant correlated with Prader scores (r(2)  = 0.253; p = 0.023). The CYP21A2 genotypes influence the severity of genital virilization in 21OHD females. We also suggest that the CAR variant, which results in a poor metabolizer phenotype, could account for a higher degree of external genitalia virilization.

BackgroundAlthough chronic adrenocorticotropic hormone (ACTH) and androgen hyperstimulation are assumed to be involved in the pathogenesis of adrenal myelolipomas associated with poor-compliance patients with congenital adrenal hyperplasia (CAH), the expression of their receptors has not yet been demonstrated in these tumors so far.MethodsWe analyzed Melanocortin 2 receptor (MC2R), Androgen Receptor (AR), Leptin (LEP), and Steroidogenic factor 1 (SF1) expression using real-time qRT-PCR in two giant bilateral adrenal myelolipomas from two untreated simple virilizing CAH cases and in two sporadic adrenal myelolipomas. In addition, the X-chromosome inactivation pattern and CAG repeat numbers in AR exon 1 gene were evaluated in the 4 cases.ResultsThe MC2R gene was overexpressed in myelolipomas from 3 out of 4 patients. AR overexpression was detected in 2 tumors: a giant bilateral myelolipoma in a CAH patient and a sporadic case. Simultaneous overexpression of AR and MC2R genes was found in two of the cases. Interestingly, the bilateral giant myelolipoma associated with CAH that had high androgen and ACTH levels but lacked MC2R and AR overexpression presented a significantly shorter AR allele compared with other tumors. In addition, X-chromosome inactivation pattern analysis showed a polyclonal origin in all tumors, suggesting a stimulatory effect as the trigger for tumor development.ConclusionThese findings are the first evidence for MC2R or AR overexpression in giant bilateral myelolipomas from poor-compliance CAH patients.

Background: The androgen receptor gene is located on the X chromosome with a polymorphic tract of CAG repeats that is inversely correlated to the receptor’s transactivation activity. A short CAG tract is associated with hyperandrogenic disorders. In women, one of the X chromosomes is inactivated and the X chromosome inactivation (XCI) pattern varies among tissues. Previous studies of hyperandrogenic disorders only evaluated XCI in leukocytes. Objective: To evaluate whether the XCI pattern in leukocytes could be extrapolated to those in hair bulbs. Material: A total of 58 healthy women were used for this study. DNA was extracted from leukocytes (n = 58 women) and pubic (n = 53 women) and scalp hair (n = 21 women). Methods:Hpa II digested and undigested DNA samples underwent fluorescence PCR GeneScan® analysis. Results: A significant and positive correlation of XCI was found between leukocytes and hair bulbs. However, individual comparisons showed that 13 and 19% of the women presented a different leukocyte XCI pattern in pubic hair and scalp hair, respectively. Conclusion: The XCI pattern was similar in leukocytes and hair bulbs of normal women indicating that leukocyte DNA is useful for XCI analysis. However, the XCI pattern could vary among tissues from the same subject, indicating that care should be taken when extrapolating individual leukocyte XCI patterns to other tissue.

INTRODUCTION:21-hydroxylase deficiency is an autosomal recessive disorder that causes glucocorticoid deficiency and increased androgen production. Treatment is based on glucocorticoid replacement; however, interindividual variability in the glucocorticoid dose required to achieve adequate hormonal control has been observed.OBJECTIVE:The present study aimed to evaluate the association between polymorphic variants involved in glucocorticoid action and/or metabolism and the mean daily glucocorticoid dose in 21-hydroxylase deficiency patients.METHODS:We evaluated 53 patients with classical forms of 21-hydroxylase deficiency who were receiving cortisone acetate. All patients were between four and six years of age and had normal androgen levels.RESULTS:The P450 oxidoreductase A503V, HSD11B1 rs12086634, and CYP3A7*1C variants were found in 19%, 11.3% and 3.8% of the patients, respectively. The mean±SD glucocorticoid dose in patients with the CYP3A7*1C and wild-type alleles was 13.9±0.8 and 19.5±3.2 mg/m2/d, respectively. We did not identify an association between the P450 oxidoreductase or HSD11B1 allelic variants and the mean glucocorticoid dose.CONCLUSION:Patients carrying the CYP3A7*1C variant required a significantly lower mean glucocorticoid dose. Indeed, the CYP3A7*1C allele accounted for 20% of the variability in the cortisone acetate dose. The analysis of genes involved in glucocorticoid metabolism may be useful in the optimization of treatment of 21-hydroxylase deficiency.

OBJECTIVE: To examine a morphokinetic approach to predicting ploidy in blastocysts by examining each of the individual morphokinetic parameters (MPs) and its utility in distinguishing euploid from aneuploid blastocysts.DESIGN: Retrospective Analysis of Morphokinetic and PGS Data. MATERIALS AND METHODS: Embryos cultured in the Embryoscope were retrospectively analyzed for MPs relative to the time of syngamy. Trophectoderm biopsy was performed and blastocysts were vitrified. Trophectoderm biopsies were analyzed using 24 chromosome array comparative genomic hybridization. MPs for each embryo were analyzed with reference to ploidy. The area under the curve (AUC) for the receiver operator characteristic (ROC) curve was used to determine whether MPs provided the ability to identify the embryo's ploidy. We focused on the two MPs (time to blastocele formation, and time to full blastocyst) used by Campbell et al. (2013) to distinguish euploid from aneuploid embryos.RESULTS: 103 embryos from 20 patients were examined. All of the 16 MPs determined in our embryos were examined using ROC curves. Most parameters, including those identified (Campbell et al., 2013) had ROCs with AUCs less than 0.6, indicating that these parameters provide poor discrimination of euploid from aneuploid embryos. Only one MP, the duration of compaction (DOC), had an AUC for its ROC curve that exceeded 0.6. Using a threshold value of 12.3 hours, we split the embryos into two groups, one with an incidence of euploidy of 55% (28/51) and the other with an incidence of euploidy of 25% (13/51). Use of Campbell's criteria provided no significant enrichment over the observed incidence of euploidy in unselected embryos.CONCLUSION: The failure of Campbell's MPs to have significant ROCs suggest that they are not universally applicable for segregating euploid from aneuploid embryos. The observation that DOC provided a significant enrichment for euploid embryos in our facility, but did not provide any enrichment in the Campbell lab (despite being considered) provides evidence that this MP is also not universal. We conclude that there is no universal MP, at present, to distinguish the ploidy of embryos. It remains unclear why MPs can yield significant, yet small, enrichment of euploidy in some facilities but are incapable of providing enrichment in other facilities.