ResearchPad - ovarian-function—from-oligomenorrhea-to-amenorrhea https://www.researchpad.co Default RSS Feed en-us © 2020 Newgen KnowledgeWorks <![CDATA[OR31-05 Emergence of Ovarian Hyperandrogenism and Luteal Insufficiency Following ESR1 Knockdown in the Mediobasal Hypothalamus of Adult Female Rhesus Monkeys]]> https://www.researchpad.co/article/elastic_article_7102 Diminished estradiol (E2) negative feedback action by neuronal ESR1 in the arcuate nucleus (ARC) of the mediobasal hypothalamus (MBH) is hypothesized to cause gonadotropin-releasing hormone (GnRH) hypersecretion, and thus LH excess, contributing to ovarian hyperandrogenism in polycystic ovary syndrome (PCOS). In primates, including humans, however, the mediating estrogen receptor is unknown. Thus, to test the hypothesis that diminished E2 action on ARC ESR1 contributes to female primate ovarian hyperandrogenism, eleven, ovary intact, adult female rhesus macaques, pair housed with female peers, received five 12µl MRI-guided MBH infusions into the rostral-to-caudal extent of both right and left ARC. Each infusion comprised gadolinium contrast agent and ~3-4 x 1010 adeno-associated virus 8 (AAV8) particles containing either a shRNA specific for ESR1 (n=6, ERaKD) or scrambled shRNA (n=5, control). Mid-surgery MRI scans identified targeting accuracy. 2-2.5 years following AAV8 infusion, EIA-determined P4 values were obtained from twice weekly serum samples; samples obtained during the follicular phase of menstrual cycles or anovulatory periods were submitted to liquid chromatography, tandem mass spectrometry (LCMS) for additional steroid hormones. LCMS-determined values were also obtained 0 hours (h) and 24 h following an IM injection of 200IU hCG. Both ERaKD (28.5 ± 1.3 days, mean ± SEM) and control (34.0 ± 3.3 days) female groups exhibited comparably regular menstrual cycles. ERaKD exhibited higher circulating levels of LH (2.8 ± 0.2 ng/ml, p=0.03), androstenedione (A4, 0.43 ± 0.03 ng/ml, p=0.03) and testosterone (T, 0.23 ± 0.03 ng/ml, p=0.09), and LH/FSH ratio (1.7 ± 0.2, p=0.05) compared to controls (LH, 2.1 ± 0.4; A4, 0.30 ± 0.05; T, 0.18 ± 0.01 ng/ml; LH/FSH 1.3 ± 0.2). Following an ovarian androgen-stimulating hCG injection, ERaKD 24-h peak levels for T (0.28 ± 0.01 ng/ml) were higher (p=0.03) compared to controls (0.21 ± 0.01 ng/ml). In addition, luteal insufficiency emerged in ERaKD females, with mean (2.4 ± 0.3 ng/ml), peak (3.6 ± 0.4 ng/ml) and area-under-the-curve (AUC, 23.2 ± 4.2 ng/ml/days) P4 values diminished compared to controls (mean, 3.6 ± 0.1, p=0.01; peak 5.7 ± 0.1 ng/ml, p=0.01; AUC, 43.7 ± 6.7 ng/ml/days, p=0.03). Taken together, these results suggest that knockdown of ARC ESR1 disrupts Gn stimulation of ovarian function, contributing to female monkey ovarian hyperandrogenism and menstrual cycle impairment emulating PCOS in women.

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<![CDATA[OR31-02 Diagnostic Criteria for Polycystic Ovary Syndrome in Adolescents: Impact on Prevalence and Longitudinal Body Mass Index Trajectories]]> https://www.researchpad.co/article/elastic_article_7092 Background: Polycystic ovary syndrome (PCOS) is characterised by oligo-anovulation (OA), hyperandrogenism (HA) and polycystic ovary morphology (PCO). While the Rotterdam criteria (defined as 2 out of 3 features) are the most widely used criteria in adults, controversy remains for the diagnostic criteria in adolescents as many PCOS features overlap with normal pubertal physiological changes. The 2018 international evidence-based PCOS guideline recommends modified Rotterdam criteria (OA and HA) in adolescents based on expert consensus. We aimed to 1) compare the prevalence of PCOS using original and modified Rotterdam criteria in an unselected adolescent cohort and 2) explore the association between diagnostic phenotypes and long-term body mass index (BMI) trajectories. Methods: 227 adolescent females of the Western Australian Pregnancy Cohort (Raine) Study undertook detailed PCOS assessment at the mean age of 15.3 years (mean age of menarche 12.4 years). Detailed anthropometric measurements were collected from birth until age 22 years. T-test was used for group BMI comparisons and longitudinal BMI was analysed using Generalised Estimating Equations with PCOS by time and PCOS phenotypes by time as interaction terms. Results: PCOS was diagnosed in 66 (29.1%) participants using original Rotterdam criteria versus 37 (16.3%) participants using modified Rotterdam criteria. Using modified Rotterdam criteria, participants with PCOS had higher mean group BMI than participants without PCOS from age 5 years onwards. Significant interaction was detected between PCOS and time (p<0.001) on longitudinal BMI gain where higher BMI gain was observed in participants with PCOS from age 14 years onwards. Only the modified criteria phenotype was significantly associated wth long-term BMI gain whereas other PCOS phenotypes had similar BMI trajectories as participants without PCOS (p<0.001). Conclusions: Our findings validate the PCOS guideline recommendation as modified Rotterdam criteria reduce over-diagnosis of PCOS in adolescents and accurately identify the phenotype at risk of long-term weight gain. The BMI trajectories of females with and without PCOS diverge from early childhood suggesting that metabolic dysfunction in PCOS commences early in the pre-pubertal period. Disclosures: Nothing to disclose. Funding: PCOS CRE scholarship and Research Training Program Scholarship awarded to CT; NHMRC Medical Research Future fund awarded to HT; National Heart Foundation Future Leader Fellowship awarded to LM; NHMRC early career fellowship awarded to AJ.

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<![CDATA[OR31-07 Increased Caloric Intake Improves Regularity of Menses and Is Associated with Increased TT<sub>3</sub> and Leptin in Exercising Women with Oligo/amenorrhea: The “REFUEL” Randomized Controlled Trial]]> https://www.researchpad.co/article/elastic_article_6307 Exercising women often fail to consume adequate energy intake relative to energy expenditure and are thus susceptible to menstrual disturbances and poor bone health secondary to energy deficiency. Ideal treatment plans are to increase energy intake to reverse energetic suppression. The purpose of this study was to determine if REFUEL, a 12-month randomized controlled trial (RCT) of increased energy intake, improves menstrual frequency and markers of energetic status in exercising women with oligo/amenorrhea. Young, exercising women with oligo/amenorrhea were randomized into two groups. The treatment group (Oligo/Amen+Cal, n=32) increased energy intake 20-40% above baseline energy needs and the Oligo/Amen Control group (n=30) maintained exercise and eating habits for the 12-month intervention. Menses was tracked throughout the intervention by menstrual calendars and daily urine samples, energetic status was assessed by body composition and total triiodothyronine (TT3) and leptin concentrations. Conditional recurrent events Cox Proportional Hazards model tested the effects of the intervention and multi-level modelling assessed relationships among variables. There was a significant group*time interaction for body mass, percent body fat, fat mass, and TT3 concentrations (p<0.03), such that Oligo/Amen+Cal women gained more body and fat mass and had a greater increase in TT3 during the study compared to Oligo/Amen Controls. Specifically, Oligo/Amen+Cal women (21.6 yrs, BMI: 20.2 kg/m2) increased energy intake by 353 kcal/d and gained 1.9 kg of body mass, corresponding to increased fat mass (1.2 kg) and leptin (64%). Oligo/Amen Controls (20.9 yrs, BMI: 21.3 kg/m2) had no change (-32 kcal/d) in energy intake (p<0.001 vs. Oligo/Amen+Cal) and minimal change in body mass (0.8 kg; p=0.04 vs. Oligo/Amen+Cal), fat mass (0.4 kg; p=0.08 vs. Oligo/Amen+Cal), and leptin (21% increase, p=0.07 vs. Oligo/Amen+Cal). Controlling for baseline BMI and menstrual status, the intervention increased the likelihood of experiencing menses (p<0.001) such that Oligo/Amen+Cal women were twice as likely (104% increase) to experience menses during the intervention compared to Oligo/Amen Controls. Further, the higher the BMI at baseline, the greater the likelihood of experiencing a menses such that for every kg/m2 increase in BMI the likelihood of menses increased by 10%. Overall, a nutritional intervention designed to increase energy intake by a moderate amount in exercising women with oligo/amenorrhea successfully improved body mass and fat mass, concentrations of metabolic hormones, and the likelihood of experiencing menses compared to oligo/amenorrheic women who maintained exercise and eating habits. As such, treatment plans designed to increase energy intake can be successful in reversing energetic suppression and recovering menses.

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<![CDATA[OR31-06 Candidate Gene Variants in a Large Cohort of Women with Primary Ovarian Insufficiency]]> https://www.researchpad.co/article/N2cd3d3e8-cdbc-4c9b-929f-bd5c8c2f5cc7

Abstract

Primary ovarian insufficiency (POI) is highly heritable. The majority of cases have no known cause. We hypothesized that mutations in previously identified genes or genes from the same pathways are the cause of POI in a recessive or dominant manner. Subjects included 294 women diagnosed with POI (amenorrhea with an elevated FSH level). All had a 46XX karyotype, and normal

repeat number. Subjects were recruited in Boston (n=95), at the NIH and Washington University (n=98), and in Pittsburgh (n=98). Controls included subjects recruited for health in old age and disorders unrelated to reproduction or cancer, and subjects from the 1000 Genomes Project (total n=587). Variants were called using the Sentieon software package (

). Case and control samples were stratified on ethnicity, relatedness and heterozygosity. Peddy and XPAT were used to calculate quality control metrics to detect outlier samples for removal from analysis to create a homogenous dataset. The number of cases (227) and controls (458) was adjusted for downstream analysis. XPAT imposed additional quality filters and removed variants. A second filter removed variants that did not pass a Gnomad filter of <0.001 allele frequency. VAAST was used to determine a composite likelihood ratio (CLR) as the test statistic to represent the aggregate burden of variants of affected individuals in each transcript relative to a set of 458 control genomes. The significance of each transcript’s VAAST CLR score was evaluated by 1 million permutations. We screened exomes for variants in previously identified genes causing POI in humans and those demonstrating infertility in a male or female mouse model. We also used the American College of Medical Genetics and Genomics standards for interpretation of pathogenicity of a variant, with priority on null variants in genes with probability of loss of function intolerance based on the observed vs. expected rate in gnomAD, in vivo or in vitro functional evidence of a damaging effect, significantly increased prevalence compared to controls, i.e. not found in any controls or in fewer than 10 in the gnomAD database if the subject had a matching race/ethnicity. Thirty-four subjects were removed for poor quality exomes and relatedness. Fifty-three subjects had at least one variant in a previously identified POI gene or one in which there was a previously identified functional model. Two subjects carried recessive variants and 30 carried at least one novel heterozygous candidate variant for follow up. Analysis of genetic causes of POI in this large cohort identified candidate causal gene variants in over half of the subjects. The data demonstrate that the genetic architecture is heterogeneous. Although recessive mutations have been identified in consanguineous families, the data suggest that a dominant or oligogenic pattern of inheritance may be important.

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<![CDATA[OR31-01 A Randomized Controlled Trial of a Lifestyle Intervention with Longitudinal Follow up on Ovarian Dysmorphology, Hyperandrogenism, and Menstrual Irregularity in Women with Polycystic Ovary Syndrome]]> https://www.researchpad.co/article/N7f8abff6-2add-42b7-ae04-14dacbd89cad <![CDATA[OR31-03 Single-Cell Profiling of Adult Human Ovarian Cortex Reveals Six Main Cell Types but No Germline Stem Cells]]> https://www.researchpad.co/article/Ne2deb5da-c179-416e-8e27-62acbf46a783 24,000 cells revealed the presence of six main cell types in ovarian cortex; oocytes, granulosa cells, immune cells, endothelial cells, perivascular cells, and stromal cells. Surface marker screening showed robust expression of 43 cell surface antigens in ovarian cortex cells. With the help of transcriptomic and cell surface antigen profiles, the DDX4 Ab+ cells were identified as perivascular cells. This finding was validated by immunostaining of ovarian tissue showing DDX4 Ab+ cells lining CD31 positive endothelial cells of blood vessels. To search for germline stem cells on a broader front, we compared our data with human fetal ovary cells including pre-meiotic germ cells (Li et al. 2017) and found no evidence for the presence of germ line stem cells of any kind in adult human ovarian cortex. In summary, we provide the first cell map of human ovarian cortex. Our results demonstrate six main cell types, but cannot provide support to the existence of oogonial stem cells. This dataset will be a valuable tool for studying the role of specific cell populations in ovarian biology, dissecting causes of infertility, and developing novel assisted reproductive technologies or even contraceptives. ]]> <![CDATA[OR31-04 Androgen Signaling Regulates Female Fertility Through Modulation of H3K27me3 in Granulosa Cells]]> https://www.researchpad.co/article/N6bb21297-2325-4828-9894-afa9b26c4e22