INTRODUCTION:Nonclassical 21-hydroxylase deficiency (NC21OHD) is a rare autosomal recessive disorder that is frequently misdiagnosed as polycystic ovary syndrome (PCOS) because of their overlapping clinical and biochemical presentations. The conventional diagnostic method, the cosyntropin stimulation test, is invasive, time consuming, and difficult to implement in routine clinical practice. To overcome these limitations, we previously developed a proof-of-concept machine learning model integrating basal serum steroid profiling by liquid chromatography with tandem mass spectrometry (LC-MS/MS) to identify NC21OHD without the need for dynamic stimulation testing. The present study aimed to validate and refine this model in a larger, multicentric cohort to evaluate its diagnostic performance in distinguishing NC21OHD from PCOS based on baseline steroid signatures. METHODS:This tricentric study included 447 women: 263 with PCOS and 59 with NC21OHD in the training set (Pitié-Salpêtrière Hospital, Paris), and 104 PCOS and 21 NC21OHD in 2 independent validation cohorts (Saint-Antoine Hospital, Paris; Bordeaux University Hospital). Twenty serum steroids were quantified by LC-MS/MS and analyzed using orthogonal partial least squares discriminant analysis. Model robustness was assessed by internal cross-validation and external validation. RESULTS:20-steroid model achieved complete separation between NC21OHD and PCOS in both external validation cohorts (accuracy = 100%). The most discriminant metabolites were 21-deoxycortisone, 21-deoxycortisol, and 17-hydroxyprogesterone. Simplified 3- and 6-steroid models demonstrated reduced sensitivity (∼90%) and specificity (∼97%). CONCLUSION:Machine learning combined with basal LC-MS/MS steroid profiling enables accurate identification of NC21OHD without dynamic stimulation. This approach could simplify diagnosis, improve patient comfort, and support large-scale screening of hyperandrogenic women.

BACKGROUND:A major limitation of newborn screening (NBS) for congenital adrenal hyperplasia (CAH) is the lack of specificity of the fluoroimmunoassay (FIA) currently used for 17-hydroxyprogesterone (17OHP) determination. This issue is more pronounced in newborns, due to elevated levels of interfering compounds. Fluoroimmunoassay at our NBS centre in Ile-de-France has a false-positive rate of around 80% and a predictive positive value of 16% for first- and second-tier measurements from dried blood spots. Recently, tandem mass spectrometry (LC-MS/MS) has gained international recognition as a complementary tool to FIA testing. Currently, the most frequently used biomarkers are 17OHP and 21-deoxycortisol, used either alone or in combination with steroid ratios such as cortisol or 4-androstenedione. Concurrently, the class of 11-oxygenate-androgens-such as 11-ketotestosterone-and more recently 11-oxygenate pregnanes-such as 21-deoxycortisone-has attracted growing interest in the diagnosis of 21-hydroxylase deficiency. These derivatives result from the combined action of 11-beta hydroxysteroid dehydrogenase and 11-beta hydroxylase. METHODS:We propose a revisited LC-MS/MS steroid profile, enriched with these classes of biomarkers, to be included in the CAH NBS algorithm. This combination could be used as a multi-steroid approach implemented using a machine learning model. RESULTS:Our preliminary results suggest that these oxygenated androgen/pregnane steroids are significantly discriminative to streamline the NBS process for CAH. We have demonstrated this in 2 different NBS centres, in the greater Paris region and in Brittany, France. CONCLUSION:This new algorithm could have an important impact on reducing the number of recall and family stress related to NBS.

Objective, Design, and Methods: Although 17-hydroxyprogesterone (17OHP) has historically been the steroid assayed in the diagnosis of congenital adrenal 21-hydroxylase deficiency (CAH-21D), its C11-hydroxylated metabolite, 21-deoxycortisol (21DF), which is strictly of adrenal origin, is assayed in parallel in this pathology. This steroid (21DF) is oxidized by 11beta-hydroxysteroid dehydrogenase type 2 into 21-deoxycortisone (21DE). In the context of CAH-21D confirmation testing, confounding factors (such as intensive care unit admission, stress, prematurity, early sampling, and variations of sex development) can interfere with the interpretation of the gold-standard biomarkers (17OHP and 21DF). Since its tissue concentrations are especially high in the placenta, we hypothesized that 21DE quantification in the neonatal periods could be an interesting biomarker in addition to 17OHP and 21DF. To verify this hypothesis, we developed a new mass spectrometry-based assay for 21DE in serum and applied it to newborns screened for CAH-21D. Results: In newborns with CAH-21D, the mean serum levels of 21DE reached 17.56 ng/mL (ranging from 8.58 ng/mL to 23.20 ng/mL), and the mean 21DE:21DF ratio was 4.99. In contrast, in newborns without CAH-21D, the 21DE serum levels were low and not statistically different from the analytical 21DE limit of quantification (0.01 ng/mL). Conclusion: Basal serum 21DE appears to be a novel sensitive and specific biomarker of CAH-21D in newborns.

Background: Advances in chromatography and mass spectrometry have allowed us to develop a novel technique for measuring intraprostatic hormone concentrations directly on prostate needle biopsies, rather than using traditional punch excision. This has significant clinical implications as intraprostatic dihydrotestosterone and testosterone levels could help monitor prostate growth, neoplasia and castration resistance. Methods: Patients undergoing radical cystoprostatectomy for bladder cancer were prospectively included. Each prostate specimen received one 90 mg punch excision and six needle biopsies. Intraprostatic hormones were dosed through gas chromatography-mass spectrometry. Results: We included twenty patients, of which eleven were incidentally diagnosed with prostate cancer; four had ISUP 1 (20%) and seven had ISUP 2 (35%). The prostate biopsy technique was unable to obtain measures for testosterone, Delta-4-androsterone and androstenedione. Tissue concentrations of DHEA, DHT, E1 and E2 can be obtained with no significant difference from the reference established on a punch from a single biopsy core sample. Conclusions: Our study demonstrates that intraprostatic concentrations of DHEA, DHT, E1 and E2 can be measured without significant difference from the reference established on a single punch excision. This finding opens the way to research on the interactions between endocrinology and prostate oncogenesis and particularly on the mechanisms of resistance to hormone therapies in vivo. Level of evidence: 2 (c) 2024 Elsevier Masson SAS. All rights are reserved, including those for text and data mining, AI training, and

Background and objective Failure rates after first-line treatment of localized prostate cancer (PCa) treatment remain high; therefore, it is essential to improve the selection and identification of at-risk patients to reduce mortality. The aim of the ANDROCAN study was to evaluate the biochemical recurrence (BCR) in patients with localized PCa treated by total prostatectomy at 5 yr after surgery, according to their presurgery gonadal status. Methods A prospective cohort study was conducted including 1318 patients undergoing total prostatectomy for localized PCa with a 5-yr postoperative follow-up. Clinical and hormonal data (assays of total testosterone [TT], bioavailable testosterone [BT], dihydrotestosterone, estrone, and estradiol were performed by gas chromatography/mass spectrometry) as well as metabolic syndrome parameters were collected at baseline before surgery. Pathological data (predominant Gleason grade 4 and stage) were collected and cross-referenced centrally. Factors associated with BCR were assessed by a multivariate analysis, and BCR-free survival was assessed by a Kaplan-Meier analysis. Key findings and limitations Among the 1318 patients, 237 had BCR of PCa. Considering demographic characteristics, populations with and without BCR were similar. However, patients with BCR had cancers with a higher Gleason score (p = 0.0001) and higher prostate-specific antigen (PSA) values (p = 0.0005) at baseline. Gleason score, pT >3a, and PSA level at baseline were positively correlated with BCR (p < 0.0001, p < 0.0001, and p = 0.0048, respectively), while BT and TT levels were not associated with BCR. This study includes patients with varying clinical characteristics, such as cancer history and metabolic syndrome, introducing variability that makes it challenging to isolate the specific effects of gonadal status on BCR. Another limitation is the lack of evaluation of long-term BCR beyond 5 yr, potentially overlooking recurrences that occur between 5 and 15 yr after surgery. This could lead to an underestimation of the actual long-term recurrence rates. Conclusions and clinical implications Overall, PSA levels, high Gleason score, and pT >3a are associated with a greater likelihood of disease recurrence following initial treatment and could serve as important prognostic indicators for predicting the risk of BCR. In this prospective study, biochemical hypogonadism was not associated with a higher occurrence of BCR within 5 yr of prostatectomy. The biological gonadal status of preoperative patients could potentially be useful for therapeutic decisions but does not provide an indication for the oncological follow-up. Patient summary Five-year follow up of patients after surgery showed that there is no association between hypogonadism (low levels of total testosterone and bioavailable testosterone) and cancer recurrence. However, cancer recurrence seems to be more associated with aggressiveness of cancer at the time of detection.

Dans le cancer de la prostate localisé, l’impact de l’hypogonadisme biochimique sur l’émergence et la progression du cancer est encore controversée. Notre objectif est de comparer les caractéristiques pathologiques et la récidive biologique (RB) à 5 ans après prostatectomie des patients atteints de cancer de la prostate (CaP) localisé en fonction du statut gonadique évalué par le total (TT) et la biodisponibilité (BT). Une étude de cohorte prospective de 1318 patients (âge 65,0, taille 174 cm, poids 81,5 kg, IMC 26,0 kg/m2, périmètre abdominal 100 cm) atteints de CaP localisé recrutés dans 4 centres urologiques en France, de 6/2013 à 6/2016 ayant tous un suivi de 5 ans postopératoire. Les paramètres du syndrome métabolique (MetS) ont été recueillis. Les dosages de TT, BT, DHT, E1 et E2 ont été réalisés par GC-MS. Un examen croisé centralisé des données pathologiques (grade de Gleason 4 prédominant (PrdGP4), stade) a été effectué. La survie sans RB a été évaluée selon Kaplan-Meier avec comparaisons par test de Log-rank. La cohorte a été divisée en 3 groupes ; le premier (n = 1067 ; 81 %) composé de patients eugonadiques dont la TT et la BT étaient normales (TT ≥ 3 ng/ml et BT ≥ 0,8 ng/ml), le deuxième (n = 251 ; 19 %) de ceux dont la TT et/ou la BT étaient diminuées et le troisième (n = 58 ; 4 %) de ceux dont la TT et la BT étaient diminuées. Les pourcentages de PrdGP4 et de pT ≥ 3 a étaient respectivement de 31 % et 30 % chez les eugonadiques et 41 % et 40 %, et 50 % et 51 % dans les deuxième et troisième groupe (différences significatives). 237 RB ont été observées (fréquence de 17 % chez les eugonadiques contre 21 % chez les hypogonadiques (différence non significative ; Fig. 1) du groupe 2 et 7 % (p = 0,017) chez ceux du groupe 3 (Fig. 1). Cette étude prospective démontre que l’hypogonadisme bioChimique a été associé à des CaP dont les caractéristiques histopathologiques sont plus fréquemment défavorables mais sans survenue significativement supérieure de RB dans les 5 ans après prostatectomie. Par conséquent, le statut gonadique biologique préopératoire des patients est utile pour la décision thérapeutique mais n’indique pas un suivi spécifique sur le plan oncologique.

STUDY QUESTION Can a combination of metabolomic signature and machine learning (ML) models distinguish nonclassic 21-hydroxylase deficiency (NC21OHD) from polycystic ovary syndrome (PCOS) without adrenocorticotrophic hormone (ACTH) testing? SUMMARY ANSWER A single sampling methodology may be an alternative to the dynamic ACTH test in order to exclude the diagnosis of NC21OHD in the presence of a clinical hyperandrogenic presentation at any time of the menstrual cycle. WHAT IS KNOWN ALREADY The clinical presentation of patients with NC21OHD is similar with that for other disorders of androgen excess. Currently, cosyntropin stimulation remains the gold standard diagnosis of NC21OHD. STUDY DESIGN, SIZE, DURATION The study was designed using a bicentric recruitment: an internal training set included 19 women with NC21OHD and 19 controls used for developing the model; a test set included 17 NC21OHD, 72 controls and 266 PCOS patients used to evaluate the performance of the diagnostic strategy thanks to an ML approach. PARTICIPANTS/MATERIALS, SETTING, METHODS Fifteen steroid species were measured in serum by liquid chromatography-mass spectrometry (LC-MS/MS). This set of 15 steroids (defined as 'steroidome') used to map the steroid biosynthesis pathway was the input for our models. MAIN RESULTS AND THE ROLE OF CHANCE From a single sample, modeling involving metabolic pathway mapping by profiling 15 circulating steroids allowed us to identify perfectly NC21OHD from a confounding PCOS population. The constructed model using baseline LC-MS/MS-acquired steroid fingerprinting successfully excluded all 17 NC21OHDs (sensitivity and specificity of 100%) from 266 PCOS from an external testing cohort of originally 549 women, without the use of ACTH testing. Blood sampling timing during the menstrual cycle phase did not impact the efficiency of our model. LIMITATIONS, REASONS FOR CAUTION The main limitations were the use of a restricted and fully prospective cohort as well as an analytical issue, as not all laboratories are equipped with mass spectrometers able to routinely measure this panel of 15 steroids. Moreover, the robustness of our model needs to be established with a larger prospective study for definitive validation in clinical practice. WIDER IMPLICATIONS OF THE FINDINGS This tool makes it possible to propose a new semiology for the management of hyperandrogenism. The model presents better diagnostic performances compared to the current reference strategy. The management of patients may be facilitated by limiting the use of ACTH tests. Finally, the modeling process allows a classification of steroid contributions to rationalize the biomarker approach and highlight some underlying pathophysiological mechanisms. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by 'Agence Française de Lutte contre le dopage' and DIM Région Ile de France. This study was supported by the French institutional PHRC 2010-AOR10032 funding source and APHP. All authors declare no competing financial interests. TRIAL REGISTRATION NUMBER N/A.

In children with premature pubarche (PP), late onset 21-hydroxylase deficiency (21-OHD), also known as nonclassical congenital adrenal hyperplasia (NCCAH), can be routinely ruled out by an adrenocorticotropic hormone (ACTH) test. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), a quantitative assay of the circulating steroidome can be obtained from a single blood sample. We hypothesized that, by applying multivariate machine learning (ML) models to basal steroid profiles and clinical parameters of 97 patients, we could distinguish children with PP from those with NCCAH, without the need for ACTH testing. Every child presenting with PP at the Trousseau Pediatric Endocrinology Unit between 2016 and 2018 had a basal and stimulated steroidome. Patients with central precocious puberty were excluded. The first set of patients (year 1, training set, n = 58), including 8 children with NCCAH verified by ACTH test and genetic analysis, was used to train the model. Subsequently, a validation set of an additional set of patients (year 2, n = 39 with 5 NCCAH) was obtained to validate our model. We designed a score based on an ML approach (orthogonal partial least squares discriminant analysis). A metabolic footprint was assigned for each patient using clinical data, bone age, and adrenal steroid levels recorded by LC-MS/MS. Supervised multivariate analysis of the training set (year 1) and validation set (year 2) was used to validate our score. Based on selected variables, the prediction score was accurate (100%) at differentiating premature pubarche from late onset 21-OHD patients. The most significant variables were 21-deoxycorticosterone, 17-hydroxyprogesterone, and 21-deoxycortisol steroids. We proposed a new test that has excellent sensitivity and specificity for the diagnosis of NCCAH, due to an ML approach.

Actuellement, il n’existe pas de références sur les concentrations d’androgènes intratissulaires prostatiques attendues chez les patients atteints de cancer de prostate. Notre objectif était de comparer les concentrations sériques et intraprostatiques des stéroïdes sexuels, auprès de patients atteints de cancer de prostate (CAP) ou d’hypertrophie bénigne de prostate (HBP). Entre septembre 2014 et janvier 2017, des hommes sélectionnés pour prostatectomie radicale pour CAP localisée ou pour adénomectomie voie haute pour HBP ont été inclus. Les échantillons sériques ont été prélevés avant chirurgie, selon les recommandations de la société internationale d’endocrinologie. Les échantillons intraprostatiques ont été prélevés à partir d’échantillons chirurgicaux frais et évalués par spectrométrie de masse en phase gazeuse, en zone centrale et périphérique. Une analyse permanova a été réalisé, ajustée sur l’âge, le volume prostatique et le PSA. Au total, 41 patients ont été inclus pour CAP et 32 patients pour HBP. Les patients présentant un CAP étaient plus jeunes, avaient des prostates moins volumineuses et un PSA plus élevé. Dans le sérum, les concentrations de testostérone totale (TT), de di-hydro-testostérone et d’oestradiol n’étaient pas significativement différentes selon la présence ou l’absence de cap. Dans le tissu prostatique, les concentrations de tt étaient significativement plus faibles (0,11 ng/mL vs 0,47 ng/mL ; p = 0,0002) et son dérivé l’oestradiol présentait des concentrations significativement plus élevées (31,0 ng/mL vs 22,3 ng/mL ; p = 0,01) dans le groupe CAP. Les concentrations de TT intraprostatiques était significativement plus faible dans la zone périphérique que dans la zone centrale pour le groupe CAP (0,07 ng/mL vs 0,15 ng/mL IC 95 % ; p = 0,001) (Tableau 1, Tableau 2). Les patients atteints de cap ont des concentrations de TT intraprostatiques inférieures aux patients atteints d’HBP. Les prostates cancéreuses semblent consommer plus de tt et produire plus d’oestradiol, notamment dans la zone périphérique.

BackgroundCurrently, there is no consensus regarding the expected concentration levels of intra‐prostatic sex steroids in patients with Prostate Cancer (PCa). Our objective was to assess the concentration levels of sex steroids in prostatic tissue and serum, in two cohorts of patients with localized PCa or benign prostatic hyperplasia (BPH).MethodsBetween September 2014 and January 2017, men selected for radical cystectomy (for bladder cancer) or open prostatectomy (for BPH), and men selected for radical prostatectomy for localized PCa were included. Blood samples were collected at baseline before surgery, and steroid concentrations were assessed following the recommendations of the Endocrine Society. Intra‐prostatic samples were collected from fresh surgical samples, and assessed by gas chromatography and mass spectrometry (GC/MS). Permanova analysis was performed. Analyses were adjusted for age, prostate weight, and prostate‐specific antigen (PSA) level.ResultsA total of 73 patients (41 patients with PCa and 32 patients with BPH) were included in this study. Patients with PCa were younger, and had smaller prostate volumes with higher levels of PSA. The levels of Total Testosterone (TT), Di‐Hydro‐Testosterone (DHT), and Estradiol (E2) in the serum were not significantly different between PCa and BPH. In PCa tissue, TT concentrations were significantly lower (0.11 ng/g vs 0.47 ng/g, P = 0.0002), however its derivative E2 had significantly higher concentrations (31.0 ng/g vs 22.3 ng/g, P = 0.01). DHT tissue concentrations were not significantly different between the two groups (5.55 ng/g vs 5.42 ng/g, P = 0.70). Intra‐prostatic TT concentrations were significantly lower in the peripheral zone than in the central zone for the CaP group (0.07 ng/g vs 0.15 ng/g, P = 0.004).ConclusionsPatients with PCa had lower intra‐prostatic TT and higher E2 concentrations levels compared to the patients with BPH. PCa seem to consume more TT and produce more E2, especially in the peripheral zone.

Background: Glycol ethers (GEs) are oxygenated solvents widely found in occupational and consumer water-based products. Some of them are well-known reproductive and developmental toxicants. Objectives: To study the variations in circulating sex steroid hormones, measured in cord blood, according to biomarkers of prenatal GE exposure. Methods: The study population comes from the PELAGIE mother-child cohort, which enrolled pregnant women from Brittany (France, 2002-2006). Maternal urine samples were collected from a random subcohort (n = 338) before 19 weeks' gestation, from which we measured 8 alkoxycarboxylic metabolites of GEs. We subsequently measured 13 sex steroid hormones and sex hormone-binding globulin (SHBG) in cord blood samples. Linear regressions adjusted for potential confounders were used, and nonlinear dose-response associations were investigated. Results: The detection rates of GE metabolites ranged from 4% to 98%; only the 5 most detected (> 20%) metabolites were investigated further. Phenoxyacetic acid (detection rate > 95%) was associated with lower levels of SHBG and various steroids (17-alpha-hydroxy-Pregnenolone, delta-5-androstenediol, and dehydroepiandrosterone) among boys and higher SHBG and 16-alpha-hydroxy-dehydroepiandrosterone levels among girls. The two other highly detected metabolites, methoxyetoxyacetic acid and butoxyacetic acid, were associated with variations in estradiol. Butoxyacetic acid was associated with higher delta-5-androstenediol levels while detectable levels of methoxyacetic acid were associated with lower levels of this hormone. Conclusion: Our study suggests that prenatal exposure to GE may affect endocrine response patterns, estimated by determining blood levels of sex steroid hormones in newborns. These results raise questions about the potential role of these changes in the pathways between prenatal GE exposure and previously reported adverse developmental outcomes, including impaired neurocognitive performance.

Failure rates after first-line treatment of localized prostate cancer (PCa) treatment remain high. Improvements to patient selection and identification of at-risk patients are central to reducing mortality. We aimed to determine if cancer aggressiveness correlates with androgen levels in patients undergoing radical prostatectomy for localized PCa. We performed a prospective, multicenter cohort study between June 2013 and June 2016, involving men with localized PCa scheduled to undergo radical prostatectomy. Clinical and hormonal patient data (testosterone deficiency, defined by total testosterone (TT) levels < 300 ng/dL and/or bioavailable testosterone (BT) levels < 80 ng/dL) were prospectively collected, along with pathological assessment of preoperative biopsy and subsequent radical prostatectomy specimens, using predominant Gleason pattern (prdGP) 3/4 grading. Of 1343 patients analyzed, 912 (68%) had prdGP3 PCa and 431 (32%) had high-grade (prdGP4, i.e., ISUP ≥ 3) disease on prostatectomy specimens. Only moderate concordance in prdGP scores between prostate biopsies and prostatectomy specimens was found. Compared with patients with prdGP3 tumors (i.e., ISUP ≤ 2), significantly more patients with prdGP4 cancers had demonstrable hypogonadism, characterized either by BT levels (17.4% vs. 10.7%, p < 0.001) or TT levels (14.2% vs. 9.7%, p = 0.020). BT levels were also lower in patients with prdGP4 tumors compared to those with prdGP3 disease. Testosterone deficiency (defined by TT and/or BT levels) was independently associated with higher PCa aggressiveness. BT is a predictive factor for prdGP4 disease, and evaluating both TT and BT to define hypogonadism is valuable in preoperative assessment of PCa (AndroCan Trial: NCT02235142).

Les concentrations intraprostatique des stéroïdes sexuels et leurs corrélations aux concentration sériques ont fait l’objet d’études parcellaire avec des méthodologies de dosage critiquables. L’objectif de l’essais STERPROSER a été rechercher des différences dans les concentrations des stéroïdes sexuels dans le plasma et des échantillons chirurgicaux frais de prostate centrale bénigne selon le volume de la prostate. Étude prospective monocentrique réalisée entre septembre 2014 et janvier 2017. L’âge, les paramètres de l’obésité et les concentrations sériques et intraprostatiques des stéroïdes sexuels ont été recueillis et mesurés par spectrométrie de masse en phase gazeuse respectant les dernières directives dans le domaine. Les calculs statistiques ont été ajustés pour l’âge et l’indice de masse corporelle (IMC). Trente-deux patients, répartis de manière égale entre les groupes de prostate de volume normal (< 50 g) et élevé (≥ 50 g), ont été inclus dans l’analyse. Les patients prostatiques à volume élevé étaient plus âgés, plus lourds et avaient un IMC plus élevé. La comparaison ajustée en fonction de l’âge et de l’IMC a montré une plus grande concentration de DHT dans les prostates de volumes élevés. Les tissus prostatiques de volumes normaux et élevés concentraient les stéroïdes sexuels d’une manière similaire. La comparaison des rapports des marqueurs substituts de l’activité enzymatique dans le tissu a mis en évidence des rapports testostérone totale/estrone et testostérone totale/estradiol similaires, et un rapport DHT/estrone plus élevé et un rapport DHT/PSA plus faible dans les prostates à volume élevé (Tableau 1, Tableau 2). L’essai STERPROSER fournit des preuves d’une activité plus élevée des enzymes 5-alpha réductase, ce qui conduit à une concentration plus élevée de DHT dans les prostates de volumes élevés. L’activité comparée de l’aromatase selon le volume de la prostate suggère une affinité enzymatique plus élevée de la 5-alpha-réductase pour la testostérone et/ou une affinité enzymatique inférieure de l’aromatase pour la testostérone dans les prostates de volumes élevés.

French and US endocrine societies recommend using GC-MS or RIA after purification (extraction + chromatography) to assess blood levels of testosterone in women. However, most of laboratories use automatized methods that have to be reserved to measure testosterone levels in men. The aim of this study was to show the consequences of analytical discrepancies of some immunological methods on the diagnostics values of testosterone levels assayed in women. Compared to GC-MS the correlations of the assayed levels varied (Spearman's rank correlation coefficients: 0.935; 0.793; 0.841; 0.852 respectively for RIA Immunotech™ with extraction and chromatographic purification; Testosterone Access-DxI800 ® ; Testosterone Immulite 2000 ® ; Testosterone II Cobas E601 ® ). The testosterone levels allowed an accurate conclusion in 95.2 %; 75.8 %; 77.4 %; 89.8 % of patients, respectively. The agreement with GC-MS results was very good for RIA method (κ=0,840), moderate for DxI800 ® method (κ=0,414), moderate for Immulite ® method

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I1 Apr 2017PD33-10 PREOPERATIVE SEX HORMONES PROFILES AND PATHOLOGICAL FEATURES OF LOCALIZED PROSTATE CANCER ARE RELATED TO BOTH TOTAL AND BIOAVAILABLE TESTOSTERONE Henry Botto, Yann Neuzillet, Marc Schneider, Morgan Rouprêt, Sarah Drouin, Marc Galiano, Xavier Cathelineau, Vincent Molinié, Camelia Radulescu, Eva Comperat, Frank Giton, Jean Fiet, Thierry Lebret, and Jean-Pierre Raynaud Henry BottoHenry Botto More articles by this author , Yann NeuzilletYann Neuzillet More articles by this author , Marc SchneiderMarc Schneider More articles by this author , Morgan RouprêtMorgan Rouprêt More articles by this author , Sarah DrouinSarah Drouin More articles by this author , Marc GalianoMarc Galiano More articles by this author , Xavier CathelineauXavier Cathelineau More articles by this author , Vincent MoliniéVincent Molinié More articles by this author , Camelia RadulescuCamelia Radulescu More articles by this author , Eva ComperatEva Comperat More articles by this author , Frank GitonFrank Giton More articles by this author , Jean FietJean Fiet More articles by this author , Thierry LebretThierry Lebret More articles by this author , and Jean-Pierre RaynaudJean-Pierre Raynaud More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.3324AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES In localized prostate cancer (PCa), impact of biochemical hypogonadism on cancer emergence and progression is still controversial. We aim to compare preoperative sex hormones plasma profiles and pathological features in localized PCa patients according to gonadal status assessed by total (TT) and bioavailable (BT). METHODS A new large prospective cohort study of 1125 (age 63.9, height 175 cm, weight 82,2 kg, BMI 26.8 30 kg/m2, waist circumference 101cm) localized PCa patients were recruited in 4 urological centre in France, from 6/2013-6/2016. Metabolic syndrome (MetS) parameters were collected. Assay of TT, BT, DHT, E1, and E2 were performed by GC-MS. A centralized cross- checked review of pathological data (Predominant Gleason pattern 4 (PrdGP4), stage) was done. RESULTS The cohort has been divided in 4 groups; the 1st group consists of patients with T and BT in the normal range (= 3 ng/ml and BT = 0,8 ng/ml), a low T in the 2nd, a low BT in the 3rd and both low T and BT in the 4th. The percentages of PrdGP4 and pT=3a were one-third in the normal gonadal patients going statistically up to one half in the hypogonadal. A 10% weight increase, due to fat, occurred in the low T patients, while no change occurred in the low BT, indicating a dichotomy in the action of T and BT. A dramatic difference in SHBG concentration between low BT and low T concentrations was observed, conferring to SHBG a key role in selecting patients at a high risk of an aggressive PCa. In fact, when considering hypogonadal patients by a threshold of TT= 3 ng/ml only, we miss 90 patients (8%) that had low BT, because of a high SHBG, no Mets but, a high %PrdG4. CONCLUSIONS Thus, a serum low BT delineate a population of aggressive PCa risk more than obesity. Consequently, for treatment decision-making, in addition to TT and obesity, BT should be assessed in the arsenal for the management of localized PCa. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e597 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Henry Botto More articles by this author Yann Neuzillet More articles by this author Marc Schneider More articles by this author Morgan Rouprêt More articles by this author Sarah Drouin More articles by this author Marc Galiano More articles by this author Xavier Cathelineau More articles by this author Vincent Molinié More articles by this author Camelia Radulescu More articles by this author Eva Comperat More articles by this author Frank Giton More articles by this author Jean Fiet More articles by this author Thierry Lebret More articles by this author Jean-Pierre Raynaud More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Dans le cancer localisé de la prostate (PCa), l’impact de l’hypogonadisme biochimique sur l’émergence du cancer et la progression est encore controversé. Nous avons comparé les profils sériques préopératoires des hormones sexuelles et les caractéristiques pathologiques chez les patients atteints de PCa localisés selon leur statut gonadique évalué par la testostéronémie totale (TT) et biodisponible (BT). Une nouvelle grande étude de cohorte prospective de 1125 patients (âge : 63,9 ans) ayant un PCa localisé, recrutés dans 4 centres urologiques en France, de 6/2013 à 6/2016. Les paramètres du syndrome métabolique (SM) ont été recueillis. Le dosage des stéroïdes sexuels ont été effectué par spectrométrie de masse en phase gazeuse. Une évaluation centralisée des données pathologiques (prédominance du grade de Gleason 4 [PrdGP4], stade) a été effectuée. La cohorte a été divisée en 4 groupes ; le 1er se composait de patients ayant une TT et BT normales (≥ 3 ng/mL et BT ≥ 0,8 ng/mL), une TT diminuée dans le 2e, une BT diminuée dans le 3e et une TT et BT diminuées dans le 4e. Les pourcentages de PrdGP4 et de pT ≥ 3a étaient d’un tiers chez les patients eugonadiques, augmentant significativement jusqu’à la moitié des hypogonadiques. Une augmentation de poids de 10 %, due à la masse grasse, était observée chez les patients à la TT diminuée, alors qu’aucun changement ne s’est produit en cas de BT diminuée, ce qui indique une dichotomie à l’action de TT et BT. Une différence significative dans la concentration de SHBG entre les patients à BT diminuée et ceux à TT diminuée a été observée, conférant à la SHBG un rôle clé dans la sélection des patients à haut risque d’un PCa agressive. En effet, lorsque les patients hypogonadiques sont définit uniquement par un seuil de TT ≥ 3 ng/mL, 90 patients (8 %) qui avaient une BT diminuée en raison d’une SHBG élevée, pas de SM, mais un taux de PrdG4 élevé étaient ignorés (Tableau 1). Une diminution de la BT définie une population à risque de PCa agressif mais ne présentant pas plus d’obésité. Par conséquent, pour la prise de décision thérapeutique, en plus de la TT et de l’obésité, la BT devrait être évaluée pour la prise en charge du PCa localisé.

Dans le cancer localisé de la prostate (PCa), l’impact des concentrations des stéroïdes sexuels sur l’agressivité du cancer est débattu, certains auteurs rapportant une association à la testostéronémie totale (TT), biodisponible (BT) ou à l’estradiol (E2). Nous avons comparé les profils sériques préopératoires des hormones sexuelles et les caractéristiques pathologiques chez les patients atteints de PCa localisés selon la prédominance du grade de Gleason 4 sur la pièce de prostatectomie. Une nouvelle grande étude de cohorte prospective de 1343 patients (âge : 63,9 ans, taille : 175 cm, poids : 82,2 kg, IMC : 26,8 30 kg/m2, circonférence abdominale : 101 cm) ayant un PCa localisé, recrutés dans 4 centres urologiques en France, de 6/2013 à 6/2016. Les paramètres du syndrome métabolique (SM) ont été recueillis. Le dosage de TT, BT, T libre (freeT), DHT, Δ5, Δ4, E1 et E2 ont été effectué par spectrométrie de masse en phase gazeuse. Une évaluation centralisée des données pathologiques (prédominance du grade de Gleason 4 [PrdGP4] stade) a été effectuée. La cohorte a été divisée en 2 groupes : – le 1er groupe se composait de patients ayant prédominance du grade 3 de Gleason [i.e. scores de Gleason ≤ 7 (3 + 4)] ; – le 2nd avait une prédominance du grade 4 de Gleason [i.e. scores de Gleason ≥ 7 (4 + 3)]. Les patients ayant les cancers les plus agressifs (PrdGP4) étaient significativement plus âgés et avaient un PSA supérieur. Les concentrations de TT et d’E2 ne différaient pas entre les deux groupes. En revanche, celles de BT et de Free T étaient inférieures chez les patients PrdGP4. Une différence significative dans la concentration de SHBG entre les patients à PrdGP4 et ceux à PrdGP3 a été observée, conférant à la SHBG un rôle clé dans la sélection des patients à haut risque d’un PCa agressif (Tableau 1). L’étude ANDROCAN, plus importante étude de cohorte prospective mondiale sur l’association entre les concentrations des stéroïdes sexuels et les caractéristiques du CaP localisés, montre qu’une diminution de la BT définie une population à risque de PCa agressif. Par conséquent, pour la prise de décision thérapeutique, en plus de la TT, la BT devrait être évaluée pour la prise en charge du PCa localisé. En revanche, l’E2 n’a pas lieu d’être prise en considération.

CONTEXT:Congenital adrenal hyperplasia (CAH) due to steroid 21-hydroxylase deficiency (CAH21) is most often diagnosed by newborn screening. The classic parameter studied is 17-hydroxy-progesterone, but the positive predictive value for the diagnosis of CAH is low in full-term newborns and even lower in preterm newborns.OBJECTIVE:To evaluate the diagnostic utility of simultaneously quantifying a large number of steroids by using liquid chromatography/tandem mass spectrometry (LC-MS/MS) from a small serum volume in patients with CAH, particularly during the neonatal period.SETTING AND PARTICIPANTS:LC-MS/MS was applied to sera from patients with CAH who had a classic form (n = 48) and rare forms (n = 2) of 21-hydroxylase deficiency, normal preterm (n = 10) and normal full-term (n = 20) neonates, and young patients without CAH (non-CAH; n = 149) but with various other diseases (delayed or advanced puberty, hirsutism, pubarche, adrenarche, simple growth retardation).METHODS:Sixteen steroids (glucocorticoids, mineralocorticoids, androgens, Δ5-steroids) were analyzed in 150 µL of serum by LC-MS/MS.RESULTS:An LC-MS/MS serum steroid profile was developed and validated to provide a reliable etiologic diagnosis of CAH. The serum levels of 17OH-progesterone and 21 deoxycortisol in non-CAH are reported, along with the rarely assayed 21-deoxycorticorticosterone and 11β hydroxy Δ4-androstenedione, which will aid in the diagnosis of CAH21. In addition, serum levels of mineralocorticoids, androgens, and Δ5-steroids allowed investigation of other forms of CAH.CONCLUSION:This steroid LC-MS/MS approach on a small serum volume is well suited for pediatrics, particularly neonatal medical practice, to aid in the diagnosis and monitoring of various forms of CAH.

BackgroundThe specific involvement of the sex steroids in the growth of the prostatic tissue remains unclear. Sex steroid concentrations in plasma and in fresh surgical samples of benign central prostate were correlated to prostate volume. MethodsMonocentric prospective study performed between September 2014 and January 2017. Age, obesity parameters, and both serum and intraprostatic concentrations of sex steroids were collected complying with the latest Endocrine Society guidelines and the steroids assessed by GC/MS. Statistical calculations were adjusted for age and body mass index (BMI). ResultsThirty-two patients, equally divided between normal- and high-volume prostate groups, were included in the analysis. High-volume prostate patients were older, heavier and had higher BMI. Comparison adjusted for age and BMI showed higher DHT concentrations in high-volume prostate. Both normal- and high-volume prostate tissues concentrate sex steroids in a similar way. Comparison of enzymatic activity surrogate marker ratios within tissue highlighted similar TT/E1 and TT/E2 ratios, and higher DHT/E1 ratio and lower DHT/PSA ratio in the high-volume prostates. ConclusionsSTERPROSER trial provides evidence for higher DHT concentration in highvolume prostates, that could reflect either higher 5-alpha reductase expression or lower expression of downstream metabolizing enzymes such as 3a-hydoxysteroid dehydrogenase.

Développer un profil stéroïdien quantitatif par spectrométrie de masse pour l’étude des déficits enzymatiques surrénaliens (DES) et ainsi substituer la LC-MS/MS aux approches analytiques de RIA. Extraction de 0,15 mL de sérum (SLE, biotage) ; HPLC (C18, phenomenex core-shell) ; LC-MS/MS (ABSiex Qtrap6500). Prématurés, nouveaux nés (< 4 semaines), enfants (< 15 ans) sans DES (DES-) ou avec déficits en 21 hydroxylase (DES21). Analyse simultanée de 15 stéroïdes : cortisol, 17OH-progestérone (17OHP), 21-désoxy-cortisol (21DF), cortisone, aldostérone, désoxycorticostérone (DOC), corticostérone, 11-désoxy-cortisol (11DF, composé S), 11β-OH-androstènedione, DHEA, 17OH-prègnénolone, progestérone, prègnénolone, delta4-androstènedione et testostérone. – Seuils de détection très inférieurs à la RIA (LOQ : 15 pg/mL pour la testostérone). – Très bonne corrélation entre LCMS et RIA notamment pour testostérone, delta4-androstènedione, cortisol et 17OHP (sauf en situation de prématurités où la 17OHP est surestimée par RIA du fait de valeurs transitoirement élevées de 17OH-prègnénolone). – Chez les prématurés, le 21DF (< 0,2 ng/mL) permet d’affirmer l’absence de DES21. – En dehors de la prématurité, le 21DF et la 17OHP sont très fortement corrélés (r = 0,89, p < 0,0001), au moment du diagnostic comme pendant le suivi thérapeutique des DES21. – La DOC, la corticostérone, l’aldostérone fortement diminués dans les DES21/DES-(p < 0,0001) suggèrent une perte de sel. À partir d’un volume de 150 μL sérum, les dosages quantitatifs simultanés de 15 stéroïdes améliorent l’étude des DES21 et des autres déficits enzymatiques surrénaliens (17-hydroxylase, 3βol-déshydrogénase, 11-hydroxylase).