Adrenal Androgen Excess Research Articles

Free testosterone and dehydroepiandrosterone sulfate serum levels in polycystic ovary syndrome women

Objectives: This study designed to detect the prevalence of ovarian and/or adrenal hyperandrogenism in women with Polycystic Ovary Syndrome (PCOS).Materials: One hundred PCOS women included in this cross sectional study after informed consent and approval of the study by local institute ethical committee. Diagnosis of PCOS based on the Rotterdam ESHRE criteria. Studied women underwent complete physical examination with calculation of body mass index (BMI) and assessment of hirsutism by modified Ferriman Gallway score. Clotted blood sample taken from studied women for measurement of serum total testosterone, sex hormone binding globulin (SHBG) and dehydroepiandrosterone sulfate (DHEAS) to detect the prevalence of ovarian and/or adrenal hyperandrogenism in PCOS women. Results: 66% of studied PCOS women presented with hirsutism, 22% of studied PCOS women presented with menstrual irregularities (hypomenorrhea, oligomenorrhea) and dysmenorrhea was the presenting symptom in 12%. Free testosterone was elevated in 50 cases of studied women; DHEAS was elevated in 30 cases of studied women while the remaining 20 cases had elevated both free testosterone and DHEAS. Mean free testosterone was 2.6±1.8 pg/ml and mean DHEAS was 5.5±3.8 mg/l. Conclusion: The ovary is the main source of excess androgen in PCOS (50% of studied women) and excess adrenal androgen found in 30% of studied PCOS women, further large studies recommended to confirm this finding.

Effects of Chronic ACTH Excess on Human Adrenal Cortex

Chronic ACTH excess leads to chronic cortisol excess, without escape phenomenon, resulting in Cushing’s syndrome. Excess adrenal androgens also occur: in females, they will overcompensate the gonadotrophic loss, inducing high testosterone; in males, they will not compensate it, inducing low testosterone. Chronic ACTH excess leads to chronic adrenal mineralocorticoid excess and low aldosterone levels: after an acute rise, aldosterone plasma levels resume low values after a few days when ACTH is prolonged. Two other mineralocorticoids in man, cortisol and 11 deoxycorticosterone (DOC), at the zona fasciculata, will not escape the long-term effect of chronic ACTH excess and their secretion rates will remain elevated in parallel. Over all, the concomitant rise in cortisol and 11 DOC will more than compensate the loss of aldosterone, and eventually create a state of chronic mineralocorticoid excess, best evidenced by the accompanying suppression of the renin plasma levels, a further contribution to the suppression of aldosterone secretion. Prolonged in vivo stimulation with ACTH leads to an increase in total adrenal protein and RNA synthesis. Cell proliferation is indicated by an increase in total DNA the resulting adrenocortical hyperplasia participates in the amplified response of the chronically stimulated gland, and the weight of each gland can be greatly increased. The growth-stimulatory effect of ACTH in vivo most likely proceeds through the activation of a local and complex network of autocrine growth factors and their own receptors; a number of compounds, including non-ACTH proopiomelanocortin peptides such as γ3-MSH, have been shown to exert some adrenocortical growth effect.

Clinical perspectives in congenital adrenal hyperplasia due to 11β-hydroxylase deficiency

Congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency is a rare autosomal recessive genetic disorder. It is caused by reduced or absent activity of 11β-hydroxylase (CYP11B1) enzyme and the resultant defects in adrenal steroidogenesis. The most common clinical features of 11 beta-hydroxylase deficiency are ambiguous genitalia, accelerated skeletal maturation and resultant short stature, peripheral precocious puberty and hyporeninemic hypokalemic hypertension. The biochemical diagnosis is based on raised serum 11-deoxycortisol and 11-deoxycorticosterone levels together with increased adrenal androgens. More than 100 mutations in CYP11B1 gene have been reported to date. The level of in-vivo activity of CYP11B1 relates to the degree of severity of 11 beta-hydroxylase deficiency. Clinical management of 11 beta-hydroxylase deficiency can pose a challenge to maintain adequate glucocorticoid dosing to suppress adrenal androgen excess while avoiding glucocorticoid-induced side effects. The long-term outcomes of clinical and surgical management are not well studied. This review article aims to collate the current available data about 11 beta-hydroxylase deficiency and its management.

Contributions of CAG repeat length in the androgen receptor gene and androgen profiles to premature pubarche in Korean girls.

The CAG repeat length of the androgen receptor (AR) gene, which exhibits an inverse relationship to AR sensitivity, might influence the development of the pubarche along with hyperandrogenemia. There are ethnic differences in the AR CAG repeat length, however, no Asian studies on premature pubarche (PP) have been reported, including Korea. Our objectives were to examine the hormone levels and AR CAG repeat length, and to assess their contributions to PP in Korean girls. Subjects with PP (n=16) and normal pubarche (NP, n=16), and normal controls (NC, n=16) were enrolled. The levels of dehydroepiandrosterone (DHEA), dehydroepiandrosterone-sulfate (DHEAS), 17-hydroxyprogesterone (17-OHP), and free testosterone (FT) were checked. The methylation-weighted (MW) average CAG repeat lengths were analyzed. The median ages at pubarche were 7.4 and 8.9 years in the PP and NP groups, respectively, and the levels of 17-OHP, DHEAS, and FT were similar in both groups. The PP group exhibited a higher DHEAS:DHEA ratio than the NP group (P=0.014). The medians of the MW average CAG repeat length of the AR gene were 22.4 for all subjects and did not differ among the PP (22.3), NP (22.4), and NC (22.2) groups. The AR CAG repeat lengths in the PP and NP groups did not correlate with DHEAS or FT levels. These results suggest that the AR CAG repeat length was not involved in the development of PP in Korean girls. However, excessive adrenal androgen levels, particularly those caused by increased sulfotransferase activity, might be important in the pathogenesis of PP.

Novel treatment strategies in congenital adrenal hyperplasia.

In recent years, important steps have been taken to improve the treatment of congenital adrenal hyperplasia (CAH), a relatively stagnant area for decades. In this review, we summarize these advances and propose future lines of investigation. The two main goals of CAH treatment are to replace the deficient hormones when necessary and to dampen the adrenocorticotropin activation and the ensuing adrenal androgen excess. Glucocorticoids have been the mainstay of CAH treatment, but available preparations only partially meet the clinical needs. Recent efforts have focused on improving the delivery of glucocorticoid replacement agents, to closer mimic the physiologic secretion pattern. Examples include modified release oral glucocorticoids and continuous subcutaneous hydrocortisone pumps. Furthermore, nonglucocorticoid approaches to address the androgen excess have emerged, such as inhibition of key androgenic enzymes and adrenocorticotropin secretion blockade by corticotropin-releasing hormone receptor antagonists. The promising recent progress made in CAH treatment brings new perspectives for individualized care in this complex disease.

Lower body weight and BMI at birth were associated with early adiposity rebound in 21-hydroxylase deficiency patients.

21-hydroxylase deficiency (21-OHD) is the most common type of congenital adrenal hyperplasia. In addition to the clinical problems caused by adrenal insufficiency and androgen excess, a risk for obesity and metabolic syndrome during young adulthood is a major ramification of the disease. Although glucocorticoid therapy is very likely to be one of the contributory factors, the precise causes of the metabolic status of adult 21-OHD patients remain to be clarified. Previously we reported that 21-OHD patients developed early onset AR, a condition which might create a risk for obesity and metabolic syndrome in adulthood. In order to elucidate the association between the onset of AR and factors during the fetal period to early infancy, we conducted a retrospective longitudinal analysis of 29 21-OHD patients (male: 14 cases, female: 15 cases, salt wasting type: 16, simple virilizing type: 13), who were identified by newborn screening and followed up at least until the age 10 years. Body size at birth, lower body weight, and lower BMI were found to precipitate the timing of AR. On the other hand, no significant association was observed between the timing of AR and sex, gestational age, treatment regimen (including cumulative dose of HDC), and disease severity (the type of the disease, the value of DHEA-S and 17-OHP). There are two points to consider: first, in 21-OHD patients treated with glucocorticoid substitution therapy, the risk for early AR cannot be reduced by adjusting the dose of glucocorticoid; second, fetal factors might affect the metabolic status of 21-OHD patients.

The next 150 years of congenital adrenal hyperplasia

Congenital adrenal hyperplasias (CAH) are a group of autosomal recessive defects in cortisol biosynthesis. Substantial progress has been made since the description of the first report, 150 years ago. This article reviews some of the recent advances in the genetics, diagnosis and treatment of CAH. In addition, we underline the aspects where further progress is required, including, among others, better diagnostic modalities for the mild phenotype and for some of the rare forms of disease, elucidation of epigenetic factors that lead to different phenotypes in patients with identical genotype and expending on treatment options for controlling the adrenal androgen excess.

Adrenal androgen excess and body mass index in polycystic ovary syndrome

Context: Adrenal hyperandrogenism affects around 25% of polycystic ovary syndrome (PCOS) patients but its relation to obesity is not totally understood. Objective: To assess dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEAS) levels in relation to body mass index (BMI) in PCOS. Design: Prospective observational study. Setting: Institutional practice at an Obstetrics/Gynecology hospital. Patients or Other Participants: This study included 136 PCOS patients, 20-35 years old, and 42 matched-age control women. The participants were classified with the BMI cutoff value of 27 kg/m2 as follows: 1) high-BMI PCOS patients; 2) low-BMI PCOS patients; 3) high-BMI control women; and 4) low-BMI control women. The data were reanalyzed with the BMI cutoff value of 30 kg/m2 to corroborate the findings in obese and non-obese patients. Intervention(s): Blood samples were taken. Main Outcome Measure(s): LH, FSH, insulin, T, androstenedione (A4), DHEA, DHEAS, and glucose levels were determined. Homeostatic model assessment was calculated. Pelvic and abdominal ultrasound for ovarian morphology and adipose tissue, respectively, were performed. Results: Obese PCOS patients presented significantly more insulin resistance than non-obese PCOS patients. The LH levels and LH/FSH ratio were significantly higher in low-BMI than in high-BMI PCOS patients. The A4 and DHEAS levels were significantly higher in non-obese than in obese PCOS patients. A significant correlation between LH and A4 in non-obese PCOS patients was observed. The frequency of hyperandrogenism by increased A4, and DHEA along with DHEAS was significantly higher in low-BMI PCOS patients compared to high-BMI PCOS patients. Some findings observed with the BMI cutoff value of 27 kg/m2 changed with the cutoff value of 30 kg/m2. Conclusions: Low BMI more than high BMI is associated with increased LH, high A4, DHEA and DHEAS levels in PCOS patients. The BMI cutoff value of 27 kg/m2 classified better than 30 kg/m2 for hormonal and metabolic characteristics.

Dexamethasone suppressibility and adrenal and ovarian venous effluents of 5α-reduced C19 conjugates in hyperandrogenic women

Several C19 conjugates, derived via 5α-reductase activity, are putative markers of peripheral androgen action and have been shown to correlate well with various clinical manifestations of androgen excess. While no ovarian vein gradient has been found for androsterone sulfate (ADT-S), androsterone glucuronide (ADT-G), 5α-androstane-3α,17β-diol sulfate (3α-diol-S), and 5α-androstane-3α,17β-diol glucuronide (3α-diol-G), the contribution of the adrenal gland to these conjugates has been unclear. Ten hirsute women were treated with 2mg/day dexamethasone (dex) for 7 days to determine the effect of adrenal androgen suppression on 5α-reduced androgen conjugate production. In addition, 11 women with mixed ovarian and adrenal androgen excess of non-neoplastic origin underwent ovarian and adrenal vein catheterization studies in order to assess gradients for the various C19 steroids. These women had significantly higher levels of both unconjugated and conjugated androgens, except for ADT-S, compared to 8 matched normal ovulatory women. After dex treatment, total testosterone (TT), unbound T (UT), androstenedione (A) and DHEAS, all decreased by 31–75%. ADT-S, ADT-G, 3α-diol-S and 3α-diol-G decreased by 48%, 71%, 46% and 68%, respectively. The suppression of the unconjugated androgens correlated highly and significantly with ADT-G. In the 11 patients undergoing adrenal venous catheterization, all patients exhibited a substantial adrenal gradient for TT and A. Of significance, in paired samples of peripheral venous and glandular effluents, no adrenal or ovarian gradient was found for any of the conjugated androgens. The data suggest that because dex suppression significantly decreases levels of the conjugated androgens, they are highly substrate dependent. However, since no adrenal or ovarian vein gradient exists, these markers of the manifestations of androgen excess largely reflect peripheral androgen metabolism.

Polycystic ovary syndrome, microbiomics and why you should be a little selfish with your time

Interview by Rona WilliamsonRicardo Azziz, MD, MPH, MBA, is Professor of Obstetrics and Gynecology (Ob/Gyn), Medicine and Medical Humanities, and President, at Georgia Regents University and CEO of the Georgia Regents Health System, Augusta, GA, USA. Georgia Regents University is one of Georgia’s four public comprehensive research universities, with recognized strengths in the health sciences, and including the state’s only academic health center. Formerly, Dr. Azziz served as Deputy Director for the Clinical and Translational Sciences Institute, Assistant Dean for Clinical and Translational Sciences, Vice-Chair of the Department of Ob/Gyn and co-Director of the Reproductive Endocrinology Fellowship Training Program at UCLA; and Director of the Center for Androgen-Related Disorders and Chair, Department of Ob/Gyn at the Cedars-Sinai Medical Center, Los Angeles, CA, USA. Dr. Azziz is an internationally recognized authority on androgen excess (disorders related to excess male hormones) in women, having published over 400 original peer-reviewed articles, book chapters and reviews. He has been funded by the NIH, among other entities, for his research for more than 25 years. Dr. Azziz’s laboratory has contributed significantly to the field of androgen excess research. This has included: establishing the populational prevalences and economic impact of the polycystic ovary syndrome (PCOS), the single most common form of androgen excess, idiopathic hirsutism and nonclassic adrenal hyperplasia, the most common form of an inherited defect of adrenal function in man; determining the familial risk for PCOS and elucidating some of the candidate genes that play a role in the disorder; identifying many of the molecular, genetic and physiologic mechanism(s) resulting in adrenal androgen excess in PCOS; and beginning to identify the unique defects in the molecular action of fat tissue in PCOS. In 2000, he received the President’s Achievement Award of the Society for Gynecologic Investigation for his research, and in that same year, he founded the Androgen Excess and PCOS Society, the only worldwide organization dedicated to promoting education and research in these disorders, where he currently serves as Senior Executive Director. His passion and focus for improving the care of women affected by reproductive endocrine disorders, and in particular androgen excess, has been recognized widely, including being named as one of, ‘America’s Top Doctors’ (Castle Connolly Medical Ltd, New York, NY, USA) from 2001 to present, and being recognized by various other lay press and patient-focused venues. Dr. Azziz’ musings on leadership, academics and healthcare can be seen at his blog site ‘Sculpting in Clay’ and now the Huffington Post .

Precursor-to-product ratios reflect biochemical phenotype in congenital adrenal hyperplasia

Precursor-to-product ratios in steroid hormone metabolism may accurately reflect enzymatic activity and production of metabolites relative to their disappearance. The purpose of this study was to explore the use of direct precursor-to-product steroid ratios to discriminate between infants with congenital adrenal hyperplasia (CAH) due to 21- α -hydroxylase deficiency and infants with no disorder, thus characterizing the biochemical phenotype in CAH. Deidentified dried blood spot samples from confirmed CAH cases identified by newborn screen (CAH-positive, N = 8) and from cases with no disorder (CAH-negative, N = 10) were obtained from the California State Newborn Screening Program. Samples (∼6.25 mm circular spots) underwent methanol and water extraction (9:1 ratio). Deuterated steroids served as isotope internal standards. 17-α-hydroxyprogesterone (17-OHP), 11-deoxycortisol (S), androstenedione (A4) and cortisol (F) concentrations were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the 17-OHP/S, 17-OHP/A4, and S/F ratios were calculated. The mean 17-OHP and A4 concentrations in samples from CAH cases were significantly increased when compared to cases with no disorder (p = 0.003 for both). 17-OHP/S and 17-OHP/A4 ratios were also significantly elevated in CAH cases (p = 0.007 and p < 0.001, respectively). In contrast, S and F concentrations and the S/F ratio were similar between the two groups. In CAH, the elevated 17-OHP/S ratio is a biomarker of diminished 21-α-hydroxylase activity, and the elevated 17-OHP/A4 ratio is a biomarker of adrenal androgen excess via increased 17,20-lyase activity. The similar S/F ratio indicates that the rate of production via 11-β-hydroxylase and disappearance of F is maintained in CAH.

What is the best approach to the teenage patient presenting with nonclassical Congenital adrenal hyperplasia: should we always treat with glucocorticoids?

Patients with Congenital adrenal hyperplasia due to partial deficiency in the enzyme 21-hydroxylase can present in childhood or adolescence with signs of adrenal androgen excess. Strategies to reduce the impact of androgen excess in females include cosmetic measures as well as antiandrogens and agents such as the combined oral contraceptive pill. Glucocorticoid may not be appropriate straightaway but can be introduced if other measures are ineffective or when pregnancy is planned.

A Giant Ovarian Cyst in a Neonate with Classical 21-Hydroxylase Deficiency with Very High Testosterone Levels Demonstrating a High-Dose Hook Effect

Congenital adrenal hyperplasia (CAH) is a group of disorders affecting the adrenal steroid synthesis. The most common form, 21-hydroxylase deficiency (21-OHD), leads to decreased production of cortisol and aldosterone with increased androgen secretion. In classic CAH, glucocorticoid treatment can be life-saving and serves to bring the symptoms under control. However, the treatment challenge is to effectively control the excess androgen effect by using the lowest possible glucocorticoid dose. Previous studies suggested a relationship between ovarian cyst formation and adrenal androgen excess, but neonatal large ovarian cysts have been very rarely reported in newborns with CAH. Here, we present the unique case of a neonate with classical 21-OHD who underwent surgery for a giant (10x8x7 cm) unilateral solitary ovarian follicular cyst on the 2nd postnatal day. Hormonal evaluation of the patient revealed high-dose hook effect for serum testosterone levels for the first time by a two-site immunoradiometric assay. Possible mechanisms by which androgen excess may cause ovarian cyst formation are discussed. Conflict of interest:None declared.

Does PCOS have developmental origins?

PRO--The similarity of reproductive and metabolic phenotype between prenatally androgenized sheep, or monkeys, and women with PCOS provides strong supportive evidence for developmental programming being important in the etiology of PCOS. CON--While one would not exclude altogether developmental determinants of PCOS, it is difficult to view PCOS as purely developmental or resulting only from intrauterine exposures or simply an adaptation gone astray.

An X-traordinary stroke

In January, 2010, a 60-year-old non-smoking, white man with mild intellectual disability, presented with acuteonset diplopia and ataxic gait. There was no consanguinity or relevant family history. CT of the brain showed a left cerebellar infarct. Blood tests showed he had polycythaemia with haemoglobin of 220 g/L (normal range 130–170). Investigations to fi nd a cause for the polycythaemia showed that although his erythropoietin concentration was normal and JAK2 mutation screening negative, serum total testosterone concentration was high at 29·1 nmol/L (normal 550 nmol/L at 30 min or 60 min), high serum concentrations of 17-hydroxyprogesterone 66·9 nmol/L (<5 nmol/L) and of androstendione, 31·6 nmol/L (2·0–9·1). His urinary steroid profi le was consistent with 21-hydroxylase defi ciency. A diagnosis of congenital adrenal hyperplasia was made. Our patient’s polycythaemia was treated with regular venesection, and glucocorticoid therapy was initiated to decrease corticotropin concentrations. Adrenal size decreased substantially within weeks of glucocorticoid initiation. The importance of glucocorticoid therapy to prevent future adrenal crises and androgen excess was emphasised. He may need testosterone therapy to maintain virilisation in the future. In September, 2010 at last follow-up he was stable. Congenital adrenal hyperplasia is the most common cause of antenatal virilisation in females. 95% of cases are due to defi ciency of 21-hydroxylase, an enzyme necessary for the synthesis of cortisol. Low serum cortisol concentrations stimulate secretion of corticotropin, which leads to excess production of androgenic steroids. Treatment is with glucocorticoids to prevent adrenal crisis and to normalise corticotropin concentrations, preventing excess androgen production. Excessive use of glucocorticoids should be avoided, because it can lead to growth retardation and iatrogenic Cushing’s syndrome. Our patient’s congenital adrenal hyperplasia was untreated for decades, and the sustained high concentrations of corticotropin caused severe adrenal hyperplasia with increased androgen production. Androgen excess during growth resulted in his short stature because of premature closure of the epiphyses. Androgen excess leads to polycythaemia through stimulation of erythropoiesis. Although polycythaemia is a well-described risk factor for stroke, androgenic polycythaemia in congenital adrenal hyperplasia has been reported only rarely. Androgen excess should be considered as a cause for poly cythaemia. Our patient’s case highlights the harmful eff ects of unfettered androgen excess in untreated congenital adrenal hyper plasia, and is also an important reminder on the importance of a genital examination as a part of a complete physical examination.

Mechanism of varied responsiveness to ACTH in patients with polycystic ovary syndrome

Objective To explore the potential mechanism of adrenal androgen excess in patients with polycystic ovary syndrome (PCOS), ACTH stimulation test was conducted and the polymorphisms in the promoter region of CYP21 A2 gene were screened to verify the variations related to the responsiveness to ACTH stimulation. Methods 30 healthy women and 101 PCOS patients, matched for age, were recruited from Ruijin hospital. Blood biochemical examinations were taken and sex hormone profiles obtained at baseline. 17 hydroxyprogesterone( 17OHP)was measured at 0 and 60 min in an ACTH stimulation test. The -710 bp -1 bp of the promoter region of CYP21A2 was sequenced in 87 PCOS patients and 30 control subjects. Results According to the post-stimulation 17 OHP levels obtained from 30 healthy women,PCOS patients were allocated into one group with high responsiveness to ACTH ( HR-PCOS, n = 21) and the other with normal responsiveness to ACTH ( NR-PCOS, n = 80). Compared with NR-PCOS subjects, HR-PCOS patients had higher testosterone( P T in the promoter region of CYP21 A2 may play a role in regulating 21 hydroxylase gene expression and further influencing 17OHP responsiveness to ACTH. Key words: Polycystic ovary syndrome; ACTH stimulation test; 17-hydroxyprogesterone; CYP21 A2; Polymorphism,single nucleotide

A Novel Point Mutation in Helix 10 of the Human Glucocorticoid Receptor Causes Generalized Glucocorticoid Resistance by Disrupting the Structure of the Ligand-Binding Domain

Generalized glucocorticoid resistance syndrome is a rare familial or sporadic condition characterized by partial insensitivity to glucocorticoids, caused by mutations in the glucocorticoid receptor (GR) gene. Most of the reported cases are adults, demonstrating symptoms associated with mineralocorticoid and/or adrenal androgen excess caused by compensatively increased secretion of the adrenocorticotropic hormone. We identified a new 2-yr-old female case of generalized glucocorticoid resistance syndrome. The patient (TJ) presented with a generalized seizure associated with hypoglycemia and hypokalemia. She also had hypertension and premature pubarche, whereas dexamethasone effectively suppressed these clinical manifestations. The patient's GR gene had a heterozygotic mutation (G-->A) at nucleotide position 2141 (exon 8), which resulted in substitution of arginine by glutamine at amino acid position 714 in the ligand-binding domain (LBD) of the GR alpha. Molecular analysis revealed that the mutant receptor had significantly impaired transactivation activity with a 2-fold reduction in affinity to ligand. It showed attenuated transactivation of the activation function (AF)-2 and reduced binding to a p160 nuclear receptor coactivator. Computer-based structural analysis revealed that replacement of arginine by glutamine at position 714 transmitted a conformational change to the LBD and the AF-2 transactivation surface, resulting in a decreased binding affinity to ligand and to the LXXLL coactivator motif. Dexamethasone treatment is effective in controlling the premature pubarche, hypoglycemia, hypertension, and hypokalemia in this child case, wherein arginine 714 plays a key role in the proper formation of the ligand-binding pocket and the AF-2 surface of the GR alpha LBD.

A p.P30L Mutation at the CYP21A2 Gene in Macedonian Patients with Nonclassical Congenital Adrenal Hyperplasia

A p.P30L Mutation at the CYP21A2 Gene in Macedonian Patients with Nonclassical Congenital Adrenal HyperplasiaNonclassical congenital adrenal hyperplasia (NCAH) is an autosomal recessive imbalance in cortisol synthesis with adrenal androgen excess. Although rarely recognized in infants, it may cause premature adrenarche and pubarche, virilization in young women and variable symptoms in young men. It is commonly caused by mutations in CYP21A2, the gene for steroid 21-hydroxylase. Patients with the p.P30L allele tend to have pronounced evidence of androgen excess but are categorized as nonclassical. We used direct molecular detection of the p.P30L mutation in CYP21A2 in 11 Macedonian NCAH patients and in 17 members of their families using polymerase chain reaction/amplification created restriction site (PCR/ACRS) analysis and digestion with restriction enzymes. The p.P30L mutation was found in a homozygous state in seven (63.6%) and in a heterozygous state in four (36.4%) patients. Of the latter, one was also heterozygous for the p.Q318X mutation. The p.P30L mutation was found in a heterozygous state in 10 (58.8%) and in a homozygous state in one (5.9%) of the family members. These findings support a role of the p.P30L mutation in NCAH.

Gene- and Environment-Dependent Neuroendocrine Etiogenesis of Homosexuality and Transsexualism

Sexual brain organization is dependent on sex hormone and neurotransmitter levels occurring during critical developmental periods. The higher the androgen levels during brain organization, caused by genetic and/or environmental factors, the higher is the biological predisposition to bi- and homosexuality or even transsexualism in females and the lower it is in males. Adrenal androgen excess, leading to heterotypical sexual orientation and/or gender role behavior in genetic females, can be caused by 21-hydroxylase deficiency, especially when associated with prenatal stress. The cortisol (F) precursor 21-deoxycortisol (21-DOF) was found to be significantly increased after ACTH stimulation in homosexual as compared to heterosexual females. 21-DOF was increased significantly before and even highly significantly after ACTH stimulation in female-to-male transsexuals. In view of these data, heterozygous and homozygous forms, respectively, of 21-hydroxylase deficiency represent a genetic predisposition to androgen-dependent development of homosexuality and transsexualism in females. Testicular androgen deficiency in prenatal life, giving rise to heterotypical sexual orientation and/or gender role behavior in genetic males, may be induced by prenatal stress and/or maternal or fetal genetic alterations. Most recently, in mothers of homosexual men--following ACTH stimulation--a significantly increased prevalence of high 21-DOF plasma values and 21-DOF/F ratios was found, which surpassed the mean + 1 SD level of heterosexual control women. In homosexual men as well--following ACTH stimulation--most of the 21-DOF plasma values and 21-DOF/F ratios also surpassed the mean + 1 SD level of heterosexual men. In only one out of 9 homosexual males, neither in his blood nor in that of his mother increased 21-DOF values and 21-DOF/F ratios were found after ACTH stimulation. In this homosexual man, however, the plasma dehydroepiandrosterone sulfate (DHEA-S) values and the DHEA-S/1000 x A (A = androstenedione) ratio were increased before and after ACTH stimulation. Furthermore, highly significantly increased basal plasma levels of dehydroepiandrosterone sulfate were found in male-to-female transsexuals as compared to normal males, suggesting partial 3 beta-ol hydroxysteroid dehydrogenase deficiency to be a predisposing factor for the development of male-to-female transsexualism.

Increased clearance of cortisol by 5β-reductase in a subgroup of women with adrenal hyperandrogenism in polycystic ovary syndrome

Increased peripheral metabolism of cortisol may explain compensatory ACTH-dependent adrenal steroidogenesis and hence hyperandrogenism in polycystic ovary syndrome (PCOS). Previous studies have described an increased 5alpha-reduction of cortisol or impaired regeneration of cortisol by 11beta-HSD1 in PCOS. However, these observations may be confounded by obesity. Moreover, the relationship between alterations in cortisol metabolism and the extent of adrenal androgen hyper-secretion in response to ACTH has not been established. This study aimed to examine the association between cortisol metabolism and ACTH-dependent adrenal hyperandrogenism in PCOS, independently of obesity. We compared 90 PCOS women (age 18-45 yr) stratified by adrenal androgen responses to ACTH1-24 and 45 controls matched for age and body weight. PCOS women were stratified as normal responders (NR), intermediate responders (IR), and high responders (HR) to 250 microg ACTH1-24: NR (no.=27) had androstenedione and DHEA responses within 2 SD of the mean in controls; IR (no.=43) had DHEA responses >2 SD above controls; HR (no.=20) had both androstenedione and DHEA responses >2 SD above controls. All groups were similar for age, body weight, and body fat distribution. Basal testosterone, androstenedione, and 5alpha-dihydrotestosterone plasma levels were similarly elevated among the 3 groups of PCOS compared with controls, whereas basal DHEA-S was higher in HR (2.8+/-1.2 microg/ml) and IR (2.4+/-1.1 microg/ml) than in NR (1.8+/-0.8 microg/ml) and controls (1.7+/-0.6 microg/ml). The HR group had the lowest basal plasma cortisol levels (101+/-36 ng/ml vs IR 135+/-42 ng/ml, NR 144+/-48 ng/ml, and controls 165+/-48 ng/ml; all p<0.01), but the greatest cortisol response to ACTH1-24 (Delta(60-0)cortisol 173+/-60 ng/ml vs IR 136+/-51 ng/ml, NR 114+/-50 ng/ml, and controls 127+/-50 ng/ml; all p<0.01), and the highest urinary excretion of total and 5beta-reduced cortisol metabolites (eg 5beta-tetrahydrocortisol/ cortisol ratio 25.2+/-15.3 vs IR 18.8+/-10.7, NR 19.7+/-11.4, and controls 17.2+/-13.7; all p<0.05). There were no differences in urinary excretion of 5alpha-reduced cortisol metabolites or in 5alpha-dihydrotestosterone/testosterone ratio between groups. Adrenal androgen excess in PCOS is associated with increased inactivation of cortisol by 5beta-reductase that may lower cortisol blood levels and stimulate ACTH-dependent steroidogenesis.