bluuebunny/crossref_metadata_2025 huggingface dataset:
FRI416 Primary Hyperaldosteronism in Pregnancy
Authors: Seav Susan M , Basina Marina
Published: October 2023

Disclosure: S.M. Seav: None. M. Basina: None.

Background: Primary hyperaldosteronism is extremely rare in pregnancy. Since 1962, only approximately 30 cases have been reported in the literature.1

Clinical Case: A 40-year-old female was referred for work-up of primary hyperaldosteronism (PHA). She was first diagnosed with hypertension (HTN) at age 29 during her first pregnancy. Laboratory work-up was consistent with PHA: serum aldosterone 22.6 ng/dL, renin activity 0.1 ng/mL/hr, aldosterone:renin ratio of 226. Post-saline infusion aldosterone level was 10.7 ng/dL. Adrenal CT scan showed a 6mm left-sided adenoma. Surgery was not recommended because adrenal vein sampling did not show lateralization. She was started on Spironolactone 25 mg/day. Despite the significant pregnancy risks, patient continued to desire conception and was switched to Labetolol. She was lost to follow-up and returned to the emergency room for dizziness at 26 weeks pregnant with twins with BP 180/109 mmHg. She had not been taking any anti-hypertensives and there was evidence of severe fetal growth restriction in both babies. After a goals of care discussion, patient deferred emergent C-section and suffered intrauterine fetal demise on hospital day 9.

Conclusion: PHA in pregnancy is rare and difficult to diagnose and manage. Newer studies estimate that > 5-10% of patients with HTN have PHA.2 Hence, maintaining high vigilance of this disease in pregnant patients is even more imperative, including screening for PHA during pre-conception period in hypertensive women. A systematic review of 38 cases of PHA in pregnancy showed that 19% of cases were diagnosed with HTN prior to pregnancy without PHA screening.1 The average age of women in this study was 29.8 years. Additionally, there is a correlation in women with younger age at diagnosis and having unilateral PHA, in which case adrenalectomy should be performed prior to conception.3 In pregnancy, first line anti-hypertensive therapies include alpha methyl dopa, labetolol, and nifedipine. In PHA, diuretics such as amiloride and mineralocorticoid receptors should also be considered if BP is not at goal. Spironolactone and eplerenone have been safely used in pregnancy with both male and female fetuses.1,4 If surgery is not an option and medical therapy is ineffective, then physicians should have a low threshold to admit patients for close BP monitoring.

1Landau, et al. Primary Aldosteronism and Pregnancy. Annales d’Endocrinologie. 2016;77:148–160 2Funder, et al. Management of Primary Aldosteronism: Case Detection, Diagnosis, and Treatment: An Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. 2016;101(5):1889–1916 3Akasaka, et al. Sex Difference in the Association Between Subtype Distribution and Age at Diagnosis in Patients With Primary Aldosteronism. Hypertension. 2019;74(2):368-374 4Cabassi, et al. Eplerenone Use in Primary Aldosteronism in Pregnancy. Hypertension. 2012;59:e18-e19

Presentation: Friday, June 16, 2023
FRI443 Optimizing Conditions Of Tubule-like Structures In Testicular Organoids That Will Support Spermatogenesis
Authors: Printy Olivia , Laronda Monica M
Published: October 2023

Disclosure: O. Printy: None.

Testicular organoids are increasingly being investigated as a potential means of fertility restoration. These organoids have been shown to self-organize to recapitulate the inner architecture of the native testis, forming tubule-like structures that resemble the seminiferous tubules. However, these tubule-like structures lack dimensionality and do not contain a large, meiotically active germ cell population. We hypothesized that if testicular organoids are cultured under appropriate microenvironmental conditions, we will observe the development of tubule-like structures with diameters that resemble what is seen in vivo, in addition to appropriate somatic cell localization and Sertoli-germ cell interactions. To test this hypothesis, we generated organoids from 5dpp male CD1 mice at three cellular seeding densities: low density (170,000 cells per well), medium density (280,000 cells per well), and high density (310,000 cells per well). The heterogenous testicular cell suspension formed compact organoids at all three seeding densities and organoid diameter increased with increasing cell number. To assess the ability of the organoids to mimic the inner architecture of a testis, we performed immunohistochemistry on 14-day-old organoids: for the major somatic cell types of the testis Leydig cells (HSD3B2), peritubular myoid cells (ACTA1), and Sertoli cells (SOX9). Thus far, only medium density organoids appear to recapitulate the compartmentalization seen in native testes. These organoids tend to contain semi-continuous ring-like structures composed exclusively of ACTA1positive cells; these structures encapsulate a population of SOX9-positive cells, while HSD3B2-expressing cells are located on the periphery of the organoids. To assess the organoids’ ability to fuse to form a larger aggregate, organoids were transferred to 3D printed troughs. However, organoids did not merge and appeared to die. However, when organoids are cultured together in microcentrifuge tubes and alginate structures, they fused to form a larger aggregate. In the future, immunohistochemistry will be used to evaluate whether any extended tubule-like structures formed within the aggregates. Additionally, spermatogonial stem cells will be injected into the aggregates and ELISA assays will be performed to assess the spermatogenic and hormonal functionality of the organoids.

Presentation: Friday, June 16, 2023
FRI448 Mother Versus Daughter Attitudes Toward Breastfeeding In American Indians: Implications For Reproductive Health Education
Authors: Seely Ellen W , Demirci Jill Radtke , Hebert Luciana E , Kelty Estelle , Heydarpour Mahyar , Moore Kelly , Charron-Prochownik Denise
Published: October 2023

Disclosure: E.W. Seely: None. J.R. Demirci: None. L.E. Hebert: None. E. Kelty: None. M. heydarpour: None. K. Moore: Consulting Fee; Self; Novo Nordisk. D. Charron-Prochownik: None.

Background: Breastfeeding has well established health benefits for mothers and their children, being associated with a reduction in incidence of type 2 diabetes and childhood overweight/obesity. Accordingly, the American Academy of Pediatrics recommends 6 months of exclusive breastfeeding, followed by 1-2 years of continued breastfeeding. American Indian and Alaska Native (AIAN) women have among the lowest rates of breastfeeding of all races/ethnicities in the United States. The low rate is striking given the high rates of type 2 diabetes and obese/overweight AIAN children and young adults. To explore the low breastfeeding rates, we examined breastfeeding attitudes among AIAN mother-daughter dyads at 5 sites (Southwest, Northeast, Plains, and Northwest Coast, U.S.) in data from the Stopping GDM randomized controlled trial (NCT02723266). Methods: We used data collected among 149 mother-daughter dyads, who were asked 7 questions from the CDC’s Infant Feeding Practices Survey II focusing on the importance of breastfeeding and its potential health benefits. An additional question was asked: “If a child was breastfed, he/she will be less likely to get diabetes.” We compared mother/daughter attitudes using Chi-square tests. Results: Mean age + standard deviation of mothers was 44.1 + 9.3 and daughters 16.7 + 3.0 years. Eighty percent of both mothers and daughters agreed or strongly agreed that “Infant formula is not as good as breast milk.” However, attitudes that the best way to feed a baby is breastfeeding and that babies should be exclusively breastfed for the first 6 months were both statistically higher in mothers than daughters (74% vs. 59% and 68% vs. 40%, respectively). Mothers also had significantly higher agreement with the statements that breastfeeding was beneficial for prevention of ear infections (66% vs. 18%), respiratory illness (67% vs. 27%), diarrhea (48% vs. 22%), obesity (42% vs. 12%) and diabetes (35% vs. 13%) than daughters. Conclusion: Significant agreement between mothers/daughters exists for the belief that infant formula is not as good as breast milk. However, daughters’ attitudes toward the benefits of breastfeeding differ significantly from mothers with daughters endorsing weaker beliefs in infant health benefits of breastfeeding. This information is important to design early interventions to increase breastfeeding in AI women by increasing awareness of benefits and eliciting maternal support for adolescents and young adult women prior to planning a pregnancy.

Presentation: Friday, June 16, 2023
FRI455 Polycystic Ovary Syndrome-Associated DENND1A Non-Coding Risk Loci Alter Gene Expression
Authors: Sankaranarayanan Laavanya , Johnson Graham , Barrera Alejandro , Dunaif Andrea Elizabeth , Reddy Timothy
Published: October 2023

Disclosure: L. Sankaranarayanan: None. G. Johnson: None. A. Barrera: None. A.E. Dunaif: None. T. Reddy: None.

Polycystic ovary syndrome (PCOS) is a leading cause of infertility and type 2 diabetes affecting 10-15% of menstruating people worldwide. PCOS is characterized by increased circulating testosterone levels, ovulatory dysfunction and insulin resistance. Genome wide association studies (GWAS) have identified ∼20 replicated common PCOS susceptibility loci in European and Han Chinese cohorts. The associated variants are predominantly non-coding and implicate neuroendocrine, reproductive, and metabolic causal pathways. Rare non-coding variants in DENND1A, a GWAS candidate gene that regulates androgen biosynthesis, were present in ∼50% of PCOS families by whole genome sequencing. These associations suggest non-coding genetic contributions to the development of PCOS. We tested the hypothesis that PCOS GWAS risk loci contribute to PCOS pathogenesis by altering gene regulation. We first identified ∼1500 functioning regulatory elements across the GWAS associated regions using a high throughput reporter assay, STARR-seq, in two different cell lines. We used H295R, an adrenal cell line model for testosterone production and COV434, an ovarian granulosa cell line. We identified cell-line specific and overlapping regulatory regions suggesting that there is context specific gene regulation. Sixty percent of the identified regions are in previously documented open chromatin regions from DNase-seq and ATAC-seq studies supporting our hypothesis that these regulatory regions are functional. We additionally experimentally tested the regulatory effect of five of the regulatory regions identified in the DENND1A locus in H295R cells using CRISPR-based epigenetic editing tools. This endogenous perturbation increases DENND1A gene levels (measured by qPCR) and also increased testosterone produced by the H295R cells (measured by ELISA), suggesting that these regions are biologically relevant to PCOS pathogenesis.We investigated allele-specific regulatory activity in a multi-ethnic pool of five genomes from healthy individuals. We identified 21 common genetic variants (minor allele frequency in the population >0.05) that alter regulatory activity in the DENND1A locus, including two that are ancestry-specific. Further, rs12237685 and rs28441318 are also eQTLS for DENND1A from the GTEX project and reside within open chromatin regions in ovarian cells. The functional consequences of non-coding genetic variants associated with complex traits has been exceptionally difficult to elucidate. Our findings suggest that both the common genetic variants and rare PCOS patient-associated DENND1A variants and altered gene expression. Further, some of those variants may contribute to allele-specific genetic risk for PCOS.

Presentation Date: Friday, June 16, 2023
FRI456 Endocrine and Non-Endocrine Causes of Fatigue in Adults With Turner Syndrome: Cohort Study and Review of the Literature
Authors: Rosenberg Anna G W , Dingemans Veerle D A , Bos-Roubos Anja G , Luijks Sanne , Dessens Arianne B , Dykgraaf Ramon , Roos-Hesselink Jolien W , van Rossum Elisabeth F C , van der Lely Aart J , de Graaff Laura C G
Published: October 2023

Disclosure: A.G. Rosenberg: None. V.D. Dingemans: None. A.G. Bos-Roubos: None. S. Luijks: None. A.B. Dessens: None. R. Dykgraaf: None. J.W. Roos-Hesselink: None. E.F. van Rossum: None. A.J. van der Lely: None. L.C. de Graaff: None.

Background: Turner syndrome (TS) is a rare genetic developmental disorder characterized by gonadal dysfunction, short stature and heart defects, among others. Women with TS often suffer from severe fatigue, for which they are typically referred to endocrinologists. The diagnostic work-up is generally time-consuming and invasive, but it rarely solves the problem. To prevent the personal and financial burden of unnecessary diagnostic procedures, it is crucial to understand fatigue in TS. Therefore, in this study, we explored the association between fatigue and endocrine and non-endocrine comorbidities in a - for rare disorders - large group of TS women. Design: Cross-sectional study. Methods: 170 genetically confirmed TS women who attended the TS reference center underwent a systematic health screening, including a structured interview, complete physical examination, biochemical measurements, perceived stress and fatigue questionnaires and additional tests when indicated. Results: Median age was 32.6 years (IQR 23.9 - 41.4 years) and median BMI was 25.4 kg/m2 (IQR 22.6 - 28.5 kg/m2). One in three TS women experienced severe fatigue. Liver enzyme disturbances and body mass index were significantly associated with higher fatigue scores, while childhood growth hormone treatment was associated with lower fatigue scores. Perceived stress was highly correlated with fatigue (ρ = 0.71; P < 0.001). Conclusions: There was no association between fatigue and most endocrine and non-endocrine disorders, which implies that fatigue is only partly explained by somatic disorders. The high correlation between perceived stress and fatigue suggests that TS-specific neuropsychological processes may play an important role in the etiology of fatigue in TS women. We provide a practical algorithm for the endocrine, non-endocrine and psychological approach to fatigue in TS women.

Presentation Date: Friday, June 16, 2023
FRI458 Rare Pathogenic Missense Variants in LMNA Identified in Women with Polycystic Ovary Syndrome (PCOS)
Authors: Bauer Rosemary , Gorsic Lidija , Legro Richard S , Ehrmann David , Geoffrey Hayes M , Urbanek Margrit
Published: October 2023

Disclosure: R. Bauer: None. L. Gorsic: None. R.S. Legro: None. M.G. Hayes: None. M. Urbanek: None.

Background: Polycystic ovary syndrome (PCOS) is the most common form of anovulatory infertility among reproductive age women. In addition to experiencing reproductive symptoms, women with PCOS are at elevated risk of developing obesity, insulin resistance (IR), and type 2 diabetes. Familial partial lipodystrophy type 2 (FPLD2) is a disorder of lipid storage and insulin resistance caused by dominant missense alleles in the gene encoding the intermediate filaments lamin A/C (LMNA). Women with this Mendelian disorder of IR also experience symptoms of PCOS such as amenorrhea and hyperandrogenism. We, therefore, hypothesize that genetic variation in LMNA also contributes to PCOS, which is a common form of IR. In other words, we hypothesize that PCOS falls into the phenotypic spectrum of disorders caused by variation in LMNA.Objective: We aim to identify and evaluate variation in LMNA that underlies PCOS pathogenesis. Methods: To test our hypothesis, we sequenced the LMNA gene in 602 women with PCOS and 125 reproductively healthy women. We comprehensively screened LMNA for genetic variation that is likely to alter the lamin A/C proteins, including missense, nonsense, splicing or frameshift variants. We identified 7 missense variants in 8 cases and no variants in reproductively healthy controls (χ2= 3.1, p=0.081, OR > 1.78, with study controls; χ2= 46.8, p<1x10-8, OR= 8.5 with gnomAD non-Finnish European cohort population controls). To determine which of these variants are pathogenic, we systematically evaluated them according to criteria outlined by the American College of Medical Genetics (ACMG). These guidelines evaluate pathogenicity of variants through multiple lines of evidence, including population data, computational predictions, and functional studies. Results: We identified 5 pathogenic variants in LMNA and 2 that are likely pathogenic. When assessed individually, 6 of the 7 variants are significantly enriched in our cohort when compared to gnomAD non-Finnish European population controls (OR > 5.0). All of the LMNA variants are likely damaging as predicted by 3 computational methods: CADD score ≥ 15, FATHMM prediction of “Damaging”, and MutationTaster classification as “disease causing.” Additionally, many of the variants have previously been identified in individuals with lipodystrophy. Conclusion: Together with previous identification of LMNA variants in women with PCOS by our lab (Urbanek et al. 2009 JCEM) and others (Crespo et al. 2022 JES), this work further establishes LMNA variation as a pathogenic mechanism for PCOS.

Presentation Date: Friday, June 16, 2023
OR19-02 DIS3 Variants Are Associated with Primary Ovarian Insufficiency
Authors: Johnstone Erica Boiman , Gorsi Bushra , Coelho Emily , Moore Barry , Chow Clement , Yandell Mark , Welt Corrine Kolka
Published: October 2023

Disclosure: E.B. Johnstone: None. B. Gorsi: None. E. Coelho: None. B. Moore: Consulting Fee; Self; Fabric Genomics. C. Chow: None. M. Yandell: Consulting Fee; Self; Fabric Genomics. Stock Owner; Self; Fabric Genomics. C.K. Welt: None.

Context: A genetic etiology accounts for the majority of unexplained primary ovarian insufficiency (POI). Objective: We hypothesized a genetic cause of POI for a sister pair with primary amenorrhea.Design: The study was an observational study.Setting: Subjects were recruited at an academic institution.Subjects: Subjects were sisters with primary amenorrhea caused by POI, and their parents. Additional subjects included women with POI analyzed previously (n=291). Controls were recruited for health in old age or were from the 1000 Genomes Project (total n=233). Intervention: We performed whole exome sequencing (WES) and data were analyzed using the Pedigree Variant Annotation, Analysis and Search Tool (pVAAST), which identifies genes harboring pathogenic variants in families. We performed functional studies in a D. melanogaster model.Main Outcome: Genes with rare pathogenic variants were identified. Results: The sisters carried compound heterozygous variants in DIS3. The sisters did not carry additional rare variants that were absent in publicly available datasets. DIS3 knockdown in the ovary of D. melanogaster resulted in lack of oocyte production and complete infertility.Conclusions: Compound heterozygous variants in highly conserved amino acids in DIS3 and failure of oocyte production in a functional model suggest that mutations in DIS3 cause POI. DIS3 is a 3’ to 5’ exoribonuclease that is the catalytic subunit of the exosome involved in RNA degradation and metabolism in the nucleus. The findings provide further evidence that mutations in genes important for transcription and translation are associated with POI.

Presentation Date: Saturday, June 17, 2023
SAT358 Race And Ethnicity Of Women With 21-hydroxylase Deficient Non-classic Adrenal Hyperplasia (NCAH) Compared With PCOS
Authors: Pace Lauren Arielle , VanHise Katherine L , Chan Jessica Loretta , Pisarska Margareta Danuta , Azziz Ricardo
Published: October 2023

Disclosure: L.A. Pace: None. K.L. VanHise: None. J.L. Chan: None. M.D. Pisarska: None. R. Azziz: Advisory Board Member; Self; Arora Forge. Consulting Fee; Self; Rani Therapeutics, Fortress Biotech, Spruce Biosciences, Core Access Surgical Technologies. Grant Recipient; Self; Foundation for Research and Education Excellence, Ferring Pharmaceuticals. Stock Owner; Self; Martin Imaging.

Background: 21-hydroxylase (21-OH) deficient non-classic adrenal hyperplasia (NCAH) is found in 1-10% of androgen excess patients and is one of the most common autosomal recessive disorders in humans. In turn, PCOS has complex inheritance and is one of the most common endocrine disorders of women. Objective: To compare the racial and ethnic distribution of NCAH vs. PCOS in a referral population. Methods: We analyzed data from a database collected prospectively of consecutive female patients presenting with androgen excess at a tertiary care referral clinic. Results: Among 645 consecutive female patients with androgen excess who completed their evaluation, 15 (2.3%) had NCAH and 619 (96.0%) had PCOS defined by the Rotterdam 2003 criteria. Of 15 women with NCAH, 13 (86.7%) were White (10 White Non-Hispanic [WNH] and 3 White Hispanic [WH]), one (6.7%) was Asian, and one (6.7%) was of mixed race. Of PCOS subjects, 479 (77.4%) were White (347 WNH, 121 WH, and 11 of undocumented ethnicity), 65 (10.5%) were Black (55 Black Non-Hispanic [BNH], 5 Black Hispanic [BH], and 5 were Black of undocumented ethnicity), 47 (7.6%) were Asian Non-Hispanic (ANH), 6 (1.0%) were Native Hawaiian/Pacific Islander Non-Hispanic, 14 (2.3%) were of mixed race (4 Hispanic and 10 Non-Hispanic), and 7 (1.1%) were other or undocumented race. Comparing key phenotypic features, the mean (±SD) age at first visit for NCAH patients was 33.1 (±14.1) years vs. 28.9 (±6.9) for PCOS (p=0.03); mean BMI of NCAH patients was 27.7 (±8.2) kg/M2 vs. 30.2 (±8.2) in PCOS (p=0.26); mean Ferriman-Gallwey (FG) score for NCAH patients was 8.6 (± 6.2) vs. 6.7 (±5.1) PCOS (p=0.19); and mean menstrual cycles per year were 7.8 (±5.2) in NCAH vs. 6.0 (±3.9) in PCOS (p=0.12). Conclusions: Phenotypically, patients with NCAH were slightly older, but did not differ in BMI, degree of hirsutism, or menstrual dysfunction vs. PCOS patients, emphasizing the need to screen for NCAH using basal follicular-phase measurements of 17-hydroxyprogesterone. However, there were notable racial differences between the NCAH and PCOS cohorts. While 10% of PCOS patients were Black in this referral clinic database, no patient with NCAH was Black. The paucity of NCAH among Black individuals evaluated for androgen excess mirrors the ethnic/racial distribution in the classic form of 21-OH deficient adrenal hyperplasia. These data question whether Black women suspected of androgen excess or PCOS should be routinely screened for NCAH.

Presentation Date: Saturday, June 17, 2023
SAT359 The Biochemical Investigation of PCOS: A UK Wide Survey of Laboratory Practice
Authors: Hawley James , Marrington Rachel , MacKenzie Finlay , Adaway Joanne , Taylor Angela , Arlt Wiebke , Keevil Brian George
Published: October 2023

Disclosure: J. Hawley: None. R. Marrington: None. F. MacKenzie: None. J. Adaway: None. A. Taylor: None. W. Arlt: None. B.G. Keevil: None.

Background Polycystic ovary syndrome (PCOS) is a heterogenous condition that affects approximately 12% of females. The diagnosis can be complex and biochemistry tests are routinely relied upon to help identify hyperandrogenaemia and to exclude other conditions. However, although national and international guidelines advocate the use biochemistry tests, little information is provided as to which tests should be used and which other endocrinopathies should be excluded. Objectives To gather information about what tests UK laboratories routinely use to investigate PCOS, what reference ranges are applied, and which other conditions are routinely excluded. Design A national survey consisting of 32 questions was compiled by clinical scientists and clinicians. This was circulated to NHS clinical laboratories via UK NEQAS and The British Endocrine Society with online access available between June – December 2021. Supplementary to the survey, UK NEQAS distributed three cases for interpretation to complement their steroid hormone scheme. Results The survey attracted responses from 81 participants. Of these, 90% identified that testosterone would be included in an initial screen with only 50% using it in combination with SHBG to provide a free androgen index. Of the conditions that are not routinely excluded; 74% would not add TSH to investigate hypothyroidism and 84% would not consider adding 17OHP to exclude late onset CAH. Testosterone analysis is commonly performed by immunoassay in the UK with only 12% of respondents using LC-MS/MS. Reference ranges for testosterone varied with the most commonly used being a manufacturer derived <1.7 nmol/L (<49 ng/dL) and the highest reported being a luteal upper limit of 6.0 nmol/L (173 ng/dL). Several participants (64%) identified that they would send high testosterones to an LC-MS/MS laboratory for confirmation, the concentration at which this occurred ranged from >1.2 nmol/L (>35 ng/dL) to >5 nmol/L (144 ng/dL). Androstenedione was only included by 16% of participants in the initial screen with the majority using LC-MS/MS for its measurement and the upper limit of normal ranging from 4.6 to 14.3 nmol/L (132 to 410 ng/dL). The results from the clinical interpretation provided varied responses. Conclusions There is significant variation across the UK in the investigation of PCOS. This is apparent in which tests are offered by biochemistry laboratories, what reference ranges are used and, as a direct consequence of these, what interpretation is applied. This potentially further complicates the investigation and diagnosis of PCOS and represents an inequality across the healthcare system. There is a requirement for clear guidance on what tests should be used to investigate PCOS.

Presentation Date: Saturday, June 17, 2023
SAT380 Clinical, Metabolic And Hormonal Profile In Women Carrying Cyp21a2 Mutations
Authors: Livadas Sarantis , Goulis Dimitrios G , Belardinelli Elisabetta , Armeni Eleni , Solmi Beatrice , Veneti Stavroula , Lambrinoudaki Irene , Cecchetti Carolina , Macut Djuro P , Gambineri Alessandra
Published: October 2023

Disclosure: S. Livadas: None. D.G. Goulis: None. E. Belardinelli: None. E. Armeni: None. B. Solmi: None. S. veneti: None. I. Lambrinoudaki: None. C. Cecchetti: None. D.P. Macut: None. A. Gambineri: None.

Background: Heterozygote (HET) CYP21A2 gene mutations are present in 5-10% of the general Mediterranean population. Despite that homozygotes suffer from classical or non-classical congenital adrenal hyperplasia (NC-CAH), a survival advantage has been suggested for heterozygote carriers. The aim of this study was to describe the clinical, metabolic and hormonal profile in reproductive age women with CYP21A2 HET. Methods: Data from 56 HET women were compared with 105 controls, 64 untreated women with NC-CAH and 63 women with polycystic ovary syndrome (PCOS) of similar age and body mass index. Results: The prevalence of hirsutism (54 vs. 8%) and acne (44 vs. 4%) was higher in HET than controls. The degree of menstrual irregularities was similar between HET and NC-CAH and worse than controls. The lipid profile was better in HET than PCOS (Cholesterol 170.2±37.1 vs. 187. 7±39.3 mg/dl, HDL 61.4±26.1 vs. 46.8±8.6 mg/dl), but similar to NC-CAH. Testosterone (0.78±0.45 vs. 0.50±0.25 ng/dl), SHBG (38.2±20.1 vs. 56.2±31.4 nmol/l), FAI (9.56±3.45 vs. 5.37±1.24) and DHEAS concentrations (2.62±1.31 vs. 1.27±0.94 nmol/l) were similar to HET, NC-CAH and PCOS and significantly higher than controls as well as insulin (13.88±7.96 vs. 7.51±2.82 IU/l) and HOMA-IR (2.95±1.60 vs. 1.65±0.67). Δ4- androstenedione (5.97±2.88 vs. 8.80±3.45 ng/ml) and 17(OH)-progesterone (4.32±2.02 vs. 13.75±8.34 ng/ml) concentrations were lower in HET than in NC-CAH, but higher than PCOS and controls. Finally, ovarian PCO morphology prevalence was similar in HET than NC-CAH and PCOS. Conclusions: HET display a favorable lipidemic profile, but a similar degree of hyperandrogenic signs, menstrual disturbances, insulin resistance, hyperandrogenemia and ovarian morphology than women with PCOS or NC-CAH. Therefore, these women, when identified, should be followed-up.

Presentation Date: Saturday, June 17, 2023
FRI176 Integrated Profiling Analysis Of Serum Steroids And Catecholamines In A Single Run With LC-MS
Authors: Noh Jongsung , Lee Chaelin , Park Seung Shin , Ahn Chang Ho , Myung Seung-Woon , Kim Jung Hee , Choi Man Ho
Published: October 2023

Disclosure: J. Noh: None. C. Lee: None. S. Park: None. C. Ahn: None. S. Myung: None. J. Kim: None. M. Choi: None.

Adrenal steroids and catecholamines are associated with the response against biochemical stress and adrenal disorders. Metabolic alteration of these hormones could be quantitatively evaluated in diagnostic process, but both adrenal cortex and medullary hormones have been measured in different analytical platforms. To increase the bio-analytical validity, liquid chromatography-mass spectrometry (LC-MS) based simultaneous profiling of 29 steroids and 6 catecholamines was developed. After alkyloxycarbonylation with isobutyl chloroformate, catecholamines and steroids were extracted simultaneously in liquid-liquid extraction. The limit of quantification ranged from 0.1 to 25 ng/mL with extraction recoveries of 78.5-97.0%, while the precision and accuracy were 4.8-7.9% and 98.0-110.9%, respectively. The validated LC-MS assay was then applied to serum samples obtained from 20 individual patients with primary aldosteronism (PA), Cushing’s syndrome (CS) and pheochromocytoma and paraganglioma (PPGL). The significantly increased serum levels of 18-hydroxycorticosterone (p < 0.005), corticosterone (p < 0.05), and 18-hydroxycortisol (p < 0.002) were observed in PA group, while PPGL group showed decreased level of 11-deoxycortisol (p < 0.007). Higher levels of glucocorticoids including 21-deoxycortisol (p < 0.002), 6β-hydroxycortisol (p < 0.001), and tetrahydrocortisone (p < 0.001) and lower levels of androgens including DHEA (p < 0.00001) and DHEA sulfate (p < 0.03) were detected in CS group. Catecholamines, such as norepinephrine (p < 0.0001) and normetanephrine (p < 0.0001), were significantly increased in PPGL patients. The present novel assay was successfully applied to discriminate three different adrenal diseases in a single analytical run.

Presentation: Friday, June 16, 2023
OR35-06 Polycystic Ovary Syndrome Is Associated With an Increased Mortality Risk
Authors: Ollila Meri-Maija , Arffman Riikka K , Morin-Papunen Laure , Korhonen Elisa , Gissler Mika , Piltonen Terhi T
Published: October 2023

Disclosure: M. Ollila: None. R.K. Arffman: None. L. Morin-Papunen: None. E. Korhonen: None. M. Gissler: None. T.T. Piltonen: None.

Women with polycystic ovary syndrome (PCOS) are burdened with multimorbidity, but there is only limited knowledge on mortality. Here, we investigated in a register-based 1:3 matched case-control study, whether women with PCOS have increased all-cause, or cause-specific mortality compared to women without PCOS. Women with PCOS (i.e., cases) were identified from the Finnish Care Register for Health Care using the International Classification of Diseases, Revisions 8, 9, and 10 (ICD-8, ICD-9, and ICD-10, respectively) codes for PCOS (ICD-10: E28.2, ICD-9: 256.4, ICD-8: 256.9). Control women were matched according to the year of birth and residential area. The time and cause of death as well as the highest level of education were extracted from the register of Statistics Finland. The registers covered the years 1969-2019. Crude and education-adjusted Cox proportional hazard models were applied, and the results were reported as hazard ratio (HR) with 95% confidence intervals. The study included 9,839 women with PCOS and 70,705 controls. Overall, 1,003 controls and 177 women with PCOS died during the follow-up period. The women with PCOS died significantly younger than the controls (51.4 ± 16.4 versus 52.9 ± 16.6 years, p<0.001). In the unadjusted analysis, women with PCOS had increased overall mortality (HR=1.53 [1.28-1.84]) as well as increased mortality due to tumors (HR=1.42 [1.06-1.90]), endocrine, nutrition, or metabolic diseases (HR=2.45 [1.02-5.96]), and diseases of the circulatory system (HR=1.77 [1.19-2.62]). In the adjusted analysis, women with PCOS had increased overall mortality (HR=1.47 [1.23-1.76]), as well as increased mortality due to diseases of the circulatory system (HR=1.67 [1.13-2.48]) and tumors (HR=1.38 [1.04-1.85]). Regarding more specific causes of death, women with PCOS had increased mortality due to diabetes (HR=3.07 [1.16-8.08]), other diseases of the circulatory system (HR=2.07 [1.00-4.25]), and bronchitis (HR=3.61 [1.01-12.88]) in the adjusted analyses. In conclusion, PCOS is a severe lifelong syndrome that increases mortality. More emphasis should be targeted on the prevention and treatment of diabetes, circulatory diseases, tumors, and respiratory diseases in women with PCOS to reduce the mortality risk.

Presentation Date: Sunday, June 18, 2023
THU574 Hsd17b1 Partially Compensates For The Lack Of Hsd17b3 In Testicular Testosterone Production In Fetal Mice, But Not In Adults
Authors: Poutanen Matti , Junnila Arttu , Sipilä Petra , Huhtaniemi Ilpo T
Published: October 2023

Disclosure: M. Poutanen: None. A. Junnila: None. P. Sipilä: None. I.T. Huhtaniemi: None.

Hydroxysteroid (17β) dehydrogenase type 3 (Hsd17b3) is the primary enzyme converting androstenedione to testosterone in human testes. HSD17B3 deficiency is a known cause of a disorder of sex development (DSD), where XY individuals are born with a female appearance, but are virilized during puberty as testosterone starts to suddenly rise due to unknown compensatory mechanisms. We have previously established and characterized an Hsd17b3 knockout mouse line (HSD17B3KO). HSD17B3KO males were born phenotypically normal, but presented various signs of under-masculinization at later ages. However, although they have very high circulating androstenedione at adulthood, they also maintain normal-to-high concentrations of testosterone. Indeed, testosterone was only seen to be low during certain fetal and pubertal time points. We suspected that Hsd17b1, capable of catalyzing the same reaction and expressed in testes towards the end of the fetal development, could be responsible for the compensation in the testosterone production, especially in the presence of higher than normal concentrations of androstenedione. To test this, we created an Hsd17b1-Hsd17b3 double KO (DKO) mouse line. The loss of both Hsd17b1 and Hsd17b3 had a more drastic effect on the phenotype of the male animals than was observed in HSD17B3KO. The outward appearance of the DKO males was completely feminized both as newborns and at adulthood, with an anogenital distance comparable to females. The male internal genitalia were present, but compared to HSD17B3KO they were further reduced in size, as were several other androgen-sensitive tissues. However, the testes and epididymides of the DKO contained high number of mature spermatozoa. The loss of both enzymes had a clear effect on the testis steroidogenesis of the newborn animals, with a 2-fold increase in intra testis androstenedione and 6-fold decrease in testosterone concentration compared to WT and HSD17B3KO males. In contrast, at the age of 3 months, no difference in the circulating steroid concentrations was observed between the DKO and HSD17B3KO, and high concentrations of androstenedione and testosterone was present in both mice. This, along with the active spermatogenesis, imply that in a presence of high LH, a further compensatory mechanism of testosterone production is present in the adult testes. In conclusion, our results demonstrate that in the absence of Hsd17b3, Hsd17b1 is crucial for the remaining testosterone production capacity in fetal mouse testis. However, in adult animals, the compensatory role is taken over by other, so far unknown, enzymes. Compared to HSD17B3KO, the overall phenotype of the DKO male mice is more similar to that of the humans with HSD17B3 deficiency, while the phenotype of HSD17B3KO is less feminized.

Presentation Date: Thursday, June 15, 2023
THU576 Anabolic Androgenic Steroid Abuse In Cis-gender Female Bodybuilder
Authors: Marta Niedzialkowska Ewelina , Shelden Daniel
Published: October 2023

Disclosure: E.M. Niedzialkowska: None. D. Shelden: None.

Introduction: Anabolic androgenic steroids (AAS) are used in bodybuilders to increase muscle mass. Abuse has surged as they are regarded as potent image enhancement drugs. There are a variety of illegal AAS available. Their true content is often unknown and dosing regimens frequently include concurrent use of multiple hormonally active agents. Use among female body builders is rarely reported. Having significantly lower levels of natural testosterone, females may be more susceptible to side effects and toxicity associated with AAS abuse. Clinical case: 36 year old cis-gender female presented to the Endocrinology clinic for evaluation of fatigue. The patient worked as a personal trainer at a gym and began participating in bodybuilding competitions. A year prior to the visit the patient was started on a steroid regimen by a friend at the gym. The patient’s regimen included testosterone cypionate (250mg-350mg a week) and trenbolone acetate. Anastozole was added to limit conversion to estrogen but was not administered on a regular basis. The patient reported no gender dysphoria. She has not had a menstrual period since she started the AAS. The patient was also diagnosed with acute kidney injury at the time of AAS initiation and further work up showed negative ANA, C3, C4, normal UA and protein/creatinine ratio. Her kidney injury was deemed to be related to NSAID and testosterone use. At the time of the visit laboratory results were significant for testosterone of >1500 ng/dl (N 240-1080), IGF-1 359 ng/ml (N 63-223), LH <0.1 mUI/ml (N>0.6), FSH <0.5 mUI/ml (N>1.4), estradiol 47 pg/ml (N 21-251), prolactin 10.8 ng/ml (N 3-30), hemoglobin 17.4 g/dl (N 12-15.1), platelets 416 bil/L (150-400), TSH 0.75 uIU/ml (N 0.4-4.5), creatinine 1.33 mg/dl (N 0.5-1.1), eGFR 53 ml/min/1.73m2. The patient's testosterone regimen was gradually decreased to 100 mg weekly with plans to further decrease the dose and with close follow up and monitoring of gonadotropins. Treatment to stimulate endogenous estrogen production may be necessary despite her current value being within normal limits as this may be a function of peripheral conversion of exogenous testosterone. Conclusion: AAS abuse is infrequent among cis-gender females. Its use in female bodybuilders increases muscle strength and athletic performance. Exogenous administration of synthetic testosterone results in negative feedback on the hypothalamic-pituitary axis, inhibiting GnRH, leading to inhibition of FSH and LH production leading to amenorrhea in females. AAS has also been associated with kidney injury with reports from animal studies suggesting testosterone induced podocyte damage and apoptosis. The steroid regimens used by bodybuilders differ and include multiple hormonally active agents. Its abuse among cis-gender women is uncommon and further research is needed to evaluate its toxicity among this group.

Presentation Date: Thursday, June 15, 2023
SAT016 Small Compounds Modulate Glucocorticoid Receptor Function By Binding To A Novel Allosteric Pocket
Authors: Jimenez-Panizo Alba , Alegre Andrea , Fuentes-Prior Pablo , Hager Gordon L , Estebanez-Perpina Eva
Published: October 2023

Disclosure: A. Jimenez-Panizo: None.

The glucocorticoid receptor (GR) is a ubiquitously expressed transcription factor that controls metabolic and homeostatic processes essential for life. This receptor exerts its functions by binding to corticosteroids through its ligand-binding domain (LBD), which harbors a hydrophobic pocket where ligands are encapsulated. Synthetic glucocorticoids, as Dexamethasone, are widely used in the clinic for the treatment of chronic inflammatory diseases. Unfortunately, their prolonged use is associated with critical side effects and most of the mechanisms underlying these collateral issues are still unknown. In this work, we present a deep biochemical and biophysical analysis showing how small molecules are able to bind a new allosteric pocket in solution and how these compounds can modulate GR transcriptional activity in cells. These findings can be essential to further understand GR signaling and how to better design synthetic corticosteroids to avoid side effects.

Presentation: Saturday, June 17, 2023
SAT021 Development Of Niclosamide Analogs For Treatment Of Androgen Receptor Positive Hepatocellular Carcinoma
Authors: Jordan Montgomery Emma , Xing Enming , Dauki Anees , Radomska Hanna , Kiyota Kulp Samuel , Granchie Lauren , Li Pui-Kai , Charles Coss Christopher
Published: October 2023

Disclosure: E.J. Montgomery: None. E. Xing: None. A. Dauki: None. H. Radomska: None. S.K. Kulp: None. L. Granchie: None. P. Li: None. C.C. Coss: None.

Introduction: Hepatocellular carcinoma (HCC) is the predominant form of liver cancer and a leading cause of cancer death worldwide. HCC is also highly sexually dimorphic with males having a 2-4 times higher risk of HCC occurrence than females regardless of the disease etiology. Based upon the discrepancy in HCC incidence between the sexes, the role of sex hormone receptors, such as the androgen receptor (AR), in HCC progression has been investigated. Despite the evidence behind AR’s role in HCC lethality, clinical trials of anti-androgen therapy in HCC showed no significant clinical benefit. We recently reported the presence of abundant ligand-independent AR splice variants (AR-SVs) in HCC. Niclosamide is an anthelmintic drug that was identified in drugs screens for AR-V7 degrading and anti-HCC properties making it well-positioned as a potential therapeutic for AR/AR-SV positive HCC. Despite these features, niclosamide suffers from poor bioavailability by design resulting in dose limiting toxicities. Objective: 1) Identify and understand key functional groups of niclosamide for development of niclosamide analogs. 2) Develop niclosamide analogs towards improving bioavailability and limiting first pass metabolism. Methods: HCC cell lines were treated with niclosamide to determine impacts on AR expression, cell proliferation, and cell invasion. Mouse pharmacokinetics studies to evaluate drug parameters were conducted through Charles River Laboratories. Toxicity studies on a valine conjugate of niclosamide were carried out in mice over a two-week span using a range of drug concentrations. Results: Numerous niclosamide analogs were evaluated resulting in a valine conjugated niclosamide with improved bioavailability, a niclosamide analog with improved first pass metabolism, and its valine conjugate. Valine conjugated niclosamide showed improved oral exposure without resulting in toxicities. Conclusions: Niclosamide is a drug showing key activity against AR and HCC that lends itself to treatment of AR/AR-SV positive HCC. Niclosamide analogs improve upon the parental drug by overcoming its limited bioavailability and its dose limiting GI toxicity while shedding light on additional pathways for further improvement.

Presentation: Saturday, June 17, 2023
OR22-04 The Oncogenic Action Of Glucocorticoid Receptor In Prostate Cancer
Authors: Paakinaho Ville
Published: October 2023

Disclosure: V. Paakinaho: None.

Steroid receptors, such as androgen (AR), and glucocorticoid receptor (GR), are hormone-controlled transcription factors (TFs) that regulate transcription in multiple central cellular systems. The action of steroid receptors has a major role in the development and progression diseases, especially endocrine-related cancers, such as prostate cancer. Since AR is the key oncogenic TF driving the development and progression of prostate cancer, the receptor is the major precision drug target in the disease. Therefore, antiandrogen treatment, i.e., the inhibition of AR activity, is a common form of cancer therapy. Despite the initial effectiveness of the therapy, antiandrogen resistance often occurs and there is no curable treatment in many cases. In contrast to AR, GR has been thought to possess tumor suppressive properties in therapy-naïve prostate cancer. Nonetheless, GR can hijack the oncogenic role of AR in prostate cancer by replacing the antiandrogen inactivated AR thus mediating antiandrogen-resistance and becoming the driver of the disease. Yet, the genome-wide effects of GR in prostate cancer have largely remained elusive. The tumor suppressive versus the oncogenic consequences of GR activation are important to acknowledge since prostate cancer patients regularly receive glucocorticoids to reduce inflammation and alleviate therapy-related side effects. To unveil these issues, we integrated ChIP-seq, RNA-seq, ATAC-seq data and uncover several pivotal features of GR action in prostate cancer cells. We detected that the replacement of AR by GR in antiandrogen-exposed prostate cancer cells occurs almost exclusively at pre-accessible chromatin sites displaying the occupancy of FOXA1. Counterintuitively to the classical pioneer factor model, the silencing of FOXA1 potentiated the chromatin binding and transcriptional activity of GR. This was attributed to FOXA1-mediated repression of the NR3C1 (gene encoding GR) expression via the corepressor TLE3. Additionally, we discovered that the chromatin binding of GR as well as transcriptional regulation by the receptor differs between different prostate cancer subtypes. Moreover, GR activity in normal prostate cells diverges drastically from that of the cancer cells. In sum, our data highlight the impact FOXA1 harbors in GR-mediated antiandrogen-resistant prostate cancer. Furthermore, the distinct GR transcriptional activity in prostate cancer subtypes could potentially be used to determine if glucocorticoid exposure possess potential harm for the prostate cancer patient.

Presentation: Saturday, June 17, 2023
OR33-06 Pioneer Factor Foxa2 Mediates Chromatin Conformation Changes In Ligand-dependent Activation Of FXR
Authors: Hao Yi , Han Lu , Wu Carol , Bochkis Irina
Published: October 2023

Disclosure: Y. Hao: None. L. Han: None. C. Wu: None. I. Bochkis: None.

Activation of nuclear receptors, a family of ligand-dependent transcription factors, is used extensively in pharmacology to develop drug targets for diverse medical conditions, including metabolic disease and cancer. We have previously shown that pioneer factor Foxa2 opens chromatin for binding of nuclear receptors FXR and LXRα during acute ligand activation (Kain et al., Molecular Metabolism, 2021). FXR is activated by bile acids and we have demonstrated that Foxa2 plays an important role in bile acid metabolism, as deletion of Foxa2 in the liver results in intrahepatic cholestasis (Bochkis et al., Nature Medicine, 2008). We hypothesized that in addition to increasing chromatin accessibility, Foxa2 also enables chromatin conformational changes during ligand activation. We performed Foxa2 HiChIP (chromatin conformation capture, Hi-C, combined with chromatin immunoprecipitation, ChIP) assay to assess Foxa2-dependent long-range interactions in mouse livers treated with vehicle and FXR agonist GW4064. HiChIP contact analysis shows that global chromatin interactions are dramatically increased during FXR activation. In addition, ligand-treated livers have more Foxa2-anchored loops, suggesting Foxa2 is involved in dynamic chromatin conformational changes in ligand-dependent FXR activation. We show that chromatin conformation, including genome-wide interactions, intra-chromosomal and inter-chromosomal loops, drastically changes upon addition of FXR agonist. Hence, Foxa2 enables these conformational changes and plays an extended role in bile acid metabolism.

Presentation: Sunday, June 18, 2023