bluuebunny/crossref_metadata_2025 huggingface dataset:
ODP422 PCOS is Associated with Increased Incidence of Co-morbidities
Authors: Vine Donna , Ghosh Mahua , Wang Grace
Published: November 2022

Background

PCOS is the most common metabolic-endocrine disorder impacting women over their lifespan. It is important to understand the scope of health outcomes that impact the health and quality of life of those with PCOS in order to improve clinical and patient-centered health care in this population.

Aim

The aim of this study was to determine the incidence of co-morbidities in PCOS compared to age matched controls. We hypothesised that women with PCOS would have increased incidence of co-morbidities.

Methods

A retrospective observational case-control study in those diagnosed with PCOS and age-matched controls (>12 yrs of age in a ratio of 1: 3) was undertaken using the Alberta Health Services Health Analytics database from 2002-2019 in Alberta, Canada. International classification codes (ICD9 and ICD10) were used to determine PCOS diagnosis including visits to a health care provider, Alberta ambulatory care reporting system and Discharge Abstract Data databases.

Results

The incidence of several co-morbidities were 20-40% higher in PCOS (n=16531) including hypertension, dyslipidemia, cardiovascular, cerebrovascular, gastrointestinal and renal disease, metabolic syndrome, eating disorders, mental illness, depression-anxiety, rheumatoid arthritis, respiratory infections and all cancers compared to controls (n=49335) (p<0. 0001). Overweight-obesity, non-alcoholic liver disease, Type 1 diabetes and Type 2 Diabetes had a 3-fold greater incidence in PCOS compared to controls (p<0. 0001).

Conclusion

These findings demonstrate PCOS is a major public health concern due to increased risk of co-morbidities and associated health-care costs. These findings provide evidence-based research to support the development of strategies to improve health care; screening, management and prevention of co-morbidities in high-risk PCOS populations.

Presentation: No date and time listed
ODP427 The Concomitant Existence of Turner Syndrome and Non-classic Congenital Adrenal Hyperplasia
Authors: Go Hanah , Joven Mark
Published: November 2022

Background

Turner syndrome and Congenital adrenal hyperplasia are distinct examples of disorders of sexual development. Because they share common clinical features, their coexistence is difficult to diagnose.1 Currently, only ten cases have been reported in the literature.2 Clinical Case: A 54-year-old, born and raised as a female, consulted a gynecologist because of ambiguous genitalia. Her late presentation was due to reluctance of medical care. During her teenage years, she had cyclic hypogastric cramps but never had menstrual bleeding. In her late 20s, she noted virilizing features such as receding hairline, hirsutism and deepening of voice. None of her parents and siblings presented the same features. She had a short stature (height: 147.32 cm), wide spaced nipples with absent breast bud and shortened 4th metatarsals. Genital examination showed a pair of labia without palpable mass and testes, and a prominent phallic structure measuring 3-4 cm (Prader Stage V). Hormonal assays revealed low normal FSH (4.58, >3. 03 mIU/mL, low LH (0.48, >1.8 mIU/mL), elevated testosterone (4.7, n<0.75 ng/mL) and progesterone (14.47, <1.4 ng/mL), and normal estradiol (66, n: 23-139 pg/mL). Tumor markers such as ß-hCG, ɑ-fetoprotein, LDH and CA-125, electrolyte analysis and thyroid function test showed no abnormalities. Both pelvic ultrasonography and abdominal MRI demonstrated a large abdominopelvic mass (14.7×17.1×8.4 cm) with no identifiable normal looking uterus, ovaries, or testes. She underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy. Histopathologic analysis of the specimen revealed submucous and intramural leiomyoma uteri, atrophic endometrium without abnormality on both ovaries and fallopian tubes. Karyotype analysis indicated 45×[2] / 46, XX [48], Mosaic 45,×/ 46 XX consistent with Turner's syndrome. Because virilization is not a typical feature, further evaluation using 17-hydroxyprogesterone was performed. The result yielded elevated basal (235, n<6 nmol/L) and postsynacthen result (60 minutes) (241, n<30 nmol/L), hence, the diagnosis of non-classic congenital adrenal hyperplasia was established.

Conclusion

The individual incidence of Turner syndrome and non-classic congenital adrenal hyperplasia is not uncommon, but their coexistence is rare. In the absence of Y chromosome in Turner syndrome, presence of virilization should warrant investigation for congenital adrenal hyperplasia.

Presentation: No date and time listed
ODP433 An increased in AIPB expression in Triple Negative (ER-/PR-/Her2-) Breast Tumors reduces estradiol synthesis
Authors: Downs William V , Perkins Edward L , Burak William E , Bose Himangshu S
Published: November 2022

Aromatase (estrogen synthetase) is a p450 family of enzymes that catalyzes testosterone to estradiol in presence of a cofactor NADPH. Aromatase is central for its role in reproduction and reproductive diseases for catalyzing the conversion of testosterone to estradiol by aromatase. Since aromatase is responsible for the synthesis of estrogens, it is essentially a rate limiting enzyme. Aromatase is expressed mainly in the ovaries of premenopausal women and placentas of pregnant women. Therapeutic approaches for controlling breast cancer proliferation include reducing estrogen by interfering with its production via ovarian ablation in premenopausal women and/or use of aromatase inhibitors or inactivators in postmenopausal women. The use of aromatase inhibitors can lead to rapid drug resistance and refractory disease. In triple negative breast cancers (TNBC) aromatase expression remains undetectable. Therefore, treatment of TNBC remains a challenge to the patients resulting in premature and very early death. Our laboratory is investigating the role of Aromatase Interacting Partner in Breast (AIPB), in TNBC cells (MDA-MB-231 cells). AIPB is a 22 kDa protein, present partially in the MAM (Mitochondria Associated-ER Membrane) but mainly in the endoplasmic reticulum associated with aromatase in the unaffected human breast tissue. The AIPB localization in the organelle was determined biochemically and visualized through scanning electron microscopy (SEM). We have cloned AIPB in a tetracycline inducible expression system (pTETOneAIPB) and created on demand expression of AIPB in MDA-MB-231 cells. In pTETOneAIPB engineered MDA-MB-231 cells addition of doxycycline resulted in a several fold increase in AIPB expression as compared to uninduced or MDA-MB-231 cells by quantitative PCR analysis. The growth rate of pTETOneAIPB is almost five-fold slower than the MBA-MB-231 cells. Interestingly estradiol synthesis was also decreased following doxycycline addition measured through radioimmuno assay (RIA). Therefore, our results suggest that restoration of AIPB expression may potentially reduce estradiol synthesis in TNBC patients and provide an additional therapeutic approached to treatment.

Presentation: No date and time listed
OR06-2 Paxillin and Androgen Receptor Interact to Modulate Female Fertility
Authors: Astapova Olga , Hammes Stephen , Vann Kenji , Walczyk Ariana , Weidner Adelaide
Published: November 2022

While androgens are traditionally associated with male physiology, recently it has become more apparent that balanced levels of androgens are crucial to maintaining female health and fertility as well. Specifically, androgens are necessary for granulosa cell proliferation and follicle survival and growth. Excess androgen levels in females are a hallmark in polycystic ovary syndrome (PCOS), whereas insufficient androgen signaling due to androgen receptor (AR) knockout leads to diminished ovarian reserve (DOR) – with both extremes resulting in anovulatory infertility. However, little is known about the factors regulating AR in granulosa cells. Our previous findings demonstrate that paxillin, a cytoplasmic adaptor protein, mediates AR signaling in prostate cancer cells. However, our current interest is whether this interaction is also found in granulosa cells. As such, our results show that paxillin indeed enhances AR protein expression in granulosa cells. We have also demonstrated through immunofluorescence and proximity ligation assays that paxillin colocalizes with AR, notably in membrane-bound complexes. Moreover, paxillin-null KGN and mouse granulosa cells express lower levels of AR, as demonstrated through Western blotting and immunofluorescence imaging. We then conducted mRNA and protein degradation experiments in paxillin-null KGN cells in order to determine the mechanism by which paxillin increases AR expression. These results suggest that paxillin acts by preventing AR mRNA and protein degradation, possibly by trafficking AR to focal adhesions, where it may be sequestered from degradation. Finally, to investigate the role of paxillin in female fertility, we developed a novel granulosa cell-specific paxillin knockout mouse model using an Amhr2-driven Cre/lox system. These knockout mice experience altered estrous cycles compared to controls, spending fewer days in estrus and more days in metestrus/diestrus. When bred continuously through 6 months of age, the paxillin KO mice produced significantly more litters compared to their littermate controls. Taken together, these results suggest that paxillin acts as a regulator of androgen-mediated female fertility in granulosa cells, and may serve as a potential target for therapeutics in fertility-related diseases such as PCOS and DOR.

Presentation: Saturday, June 11, 2022 11:45 a.m. - 12:00 p.m.
PMON227 Rare ovarian luteinized fibrothecoma as cause of severe hyperandrogenism in a pre-menopausal woman.
Authors: Mikkilineni Pushyami , Binutu Oluwabusola , Sutton Gregory , McVinnie David , Shill Jessica
Published: November 2022

Ovarian fibromas and fibrothecomas constitute up to 4.7% of ovarian sex cord stromal tumors (OSCSTs). Luteinized fibrothecomas (LFs) are a histological subvariant of OSCSTs. Typically benign and rare, LFs are mostly diagnosed in post-menopausal women with only 30% in women <30. Only 11% of LFs present with androgen excess (AE). We describe a case of severe hyperandrogenism from an ovarian LF in a young woman with primary amenorrhea and infertility.A 23-year-old woman presented for evaluation of hyperandrogenism. Iatrogenic menarche occurred at age 22, induced by letrozole and medroxyprogesterone (MP). BP was 133/86 mm Hg and BMI was 52 kg/m2. Exam revealed diffuse obesity, severe hirsutism (Ferriman Gallwey score 29), and severe acanthosis nigricans, but no scalp alopecia, deep voice, violaceous striae or bruising. Total testosterone (TT) was 108 ng/dl (2-45), free testosterone (FT) 24.7 pg/ml (0.1-6.4 dialysis), prolactin 13.2 ng/ml (<30), DHEAS 2308 ng/ml (240-4330), and TSH 1.29 µIU/mL (0.45-5.33). FSH, LH and estradiol could not be interpreted due to unclear timing in cycle. 17-hydroxyprogesterone after MP challenge was 73 ng/dl (<200). Pelvic ultrasound (US) gave normal endometrial stripe of 3.82 mm. Pelvic US and MRI showed polycystic ovarian morphology without masses. CT abdomen showed normal adrenal glands. Ovarian and adrenal vein sampling (OAVS) lateralized AE to the left ovary (LO). LO TT was 5743 ng/dl with associated IVC sample of 139 ng/dl, ratio of 41, and right ovarian TT was 381 ng/dl with associated IVC sample of 32 ng/dl, ratio of 1.18. Just prior to left oophorectomy, preoperative TT was 103 ng/dl, FT 27.8 pg/ml (0.1-6.4 dialysis), and androstenedione (A4) 212 ng/dl (mid-follicular 51-213). Two days post-operatively, TT dropped to 55 ng/dl, FT 11 pg/ml (0.2-5), and A4 105 ng/dl. Pathology revealed a 3.8 cm LF. Six weeks post-operatively, TT and FT normalized to 18 and 3.6 (0.2-5), respectively.Features atypical for PCOS, including presentation outside age 15-25, rapid progression, TT >2× upper limit of normal (ULN), and free androgen index >4× ULN should prompt thorough evaluation. Adherence to guidelines for TT threshold >150 in a pre-menopausal woman and focus on PCO morphology would have provided false reassurance. Maintaining a high degree of suspicion for an androgen-secreting tumor (AST) is paramount when clinical presentation is atypical. OAVS is usually performed when imaging fails to reveal an abnormality, as small ovarian tumors cannot be excluded. Our patient presented with symptoms atypical for PCOS including severe hirsutism and TT >2× ULN, raising concern for an AST. OAVS lateralized AE to her LO. Resection resulted in testosterone normalization and finding of a rare LF. Identifying the source of AE is especially important in women of reproductive age to restore normal reproductive function.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
ODP064 Latrogenic Cushing Syndrome after Concomitant Oral Controlled-Release Budesonide and Famotidine Therapy
Authors: Salhah Hiba , Bowden Sasigarn
Published: November 2022

Introduction

It has been shown that complement factors are increased in polycystic ovary syndrome (PCOS) and that these may be affected by obesity and insulin resistance. Hypothetically, this may increase cardiometabolic risk but, paradoxically, increased clotting is not a feature of PCOS. To investigate this, proteomic analysis of the complement system was undertaken, including inhibitory proteins.

Methods

Plasma was collected from 244 women (147 with PCOS and all 3 diagnostic features and 97 controls). Somalogic proteomic analysis was undertaken for the following complement system proteins: C1q, C1r, C2, C3, C3a, iC3b, C3b, C3d, C3adesArg, C4, C4a, C4b, C5, C5a, C5b-6 complex, C8, properdin, Factor B, Factor D, Factor H, Factor I, Mannose-binding protein C (MBL), Mannan-binding lectin serine protease 1 (MASP3), Complement decay-accelerating factor (DAF) and Complement factor H-related protein 5 (CFHR5).

Results

The alternative pathway of the complement system was primarily activated in PCOS, with increased C3 (p<0. 05), properdin and Factor B (p<0. 01). In addition, inhibition of this pathway was also seen in PCOS, with increase in CFHR5, Factor H and Factor I (p<0. 01). There were also increases in the downstream complement factors of iC3b and C3d, associated with an enhanced B cell response, and C5a, associated with inflammatory cytokine release (p<0. 01).

Conclusion

This is the most comprehensive evaluation of the complement system in PCOS to date and reveals upregulation of the alternative complement system that appears to be offset by the concurrent upregulation of its inhibitors. However, any additional dysregulation of the system, such as occurs with COVID19 infection, may give rise to an increased risk for clotting in women with PCOS.

Presentation: No date and time listed
PMON257 Oral Ingestion of Macronutrients Selectively and Variably Decrease Circulating Testosterone Concentration in Healthy Men
Authors: Iranmanesh Ali , Johannes Veldhuis
Published: November 2022

We have previously reported acute suppression of testosterone in men after oral ingestion of dextrose. In the present study, we aimed to explore a potentially similar effect in response to other macronutrients.

Eleven healthy men (age: 33-70 yrs, BMI 20.4-31.5 kg/m2) were studied in random order after an overnight fast on 4 separate days, involving oral ingestion of 300 ml of either water (control), dextrose, protein, or lipid solutions. Sessions were 6.5 h long, starting at 0800-0900 hrs. Test meals were isocaloric (400 kcal) and were ingested after the 4th blood draw (30 min point). Blood was collected at 10-min intervals for testosterone (ng/dL), and LH (IU/L) measurements. Means of the 1st 4 pre-intervention measures were used as the first value in all the time series. Mean delta-serum (macronutrients-water) testosterone and LH concentrations sampled at 10-minute intervals over 6 hour period were adjusted to the corresponding 1st value to correct for among the sessions inconsistent baseline testosterone and LH measures.

Compared to the water day (control), mean (± SE) serum testosterone concentrations (ng/dL) in 11 subjects decreased from 415 ± 36 (control) to 370 ± 31 (dextrose: P=0.009), 340 ± 28 (protein: p=0.002), and 355 ± 30 (lipid: p=0.01). Corresponding LH (IU/L) values for dextrose (3.16 ± 0.29), protein (3.17 ± 0.39), and lipid (3.07 ± 0.29) were not significantly different from the control day (3.3 ± 0.29). Mean delta-testosterone time series revealed decreased testosterone of 63 ng/dL at 130 min after dextrose, 80 ng/dL at 200 min after protein, and 10 ng/dL at 70 min after lipid ingestion. Duration of suppression was variable among the 3 macronutrients, but most prolonged for protein lasting for 320 min. As oppose to testosterone, delta-LH concentrations were more positive rather than negative, suggestive of a potential testicular insult.

In conclusion, oral ingestion of all macronutrients decrease circulating testosterone concentration, which appear to be independent of LH secretion. The effect is more intense and prolonged after protein intake, and minimal with lipid ingestion. Future studies are required to define the etiology, and the macronutrient effect on more prolonged basis.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON262 Systematic review and critical analysis on dietary supplements for male infertility: from a blend of ingredients to a rationale strategy
Authors: Grande Giuseppe , Petre Gabriel Cosmin , Pesenti Francesco Francini , Toni Luca D , Vitagliano Amerigo , Nisio Andrea D , Ferlin Alberto , Foresta Carlo , Garolla Andrea
Published: November 2022

The aim of this study was to evaluate the potential efficacy of each Dietary supplement (DS) proposed for male infertility and marketed in Italy, using an adapted version of the scoring system by the American Heart Association. In particular, the effect of any ingredients was evaluated according to semen alterations.

On the website of Italian ministry of health we found 23 supplements marketed for male infertility. We considered as eligible only randomized clinical trials (RCTs) evaluating substances included in DS marketed for male infertility. A formula allowed us to classify the expected efficacy of each DS, based on composition. Each DS was scored and included into three classes of expected efficacy: high, low and none.

Among 24 supplements,3 (12,5%) fall in high, 9 (37,5%) in lower and 12 (50,0%) in no expected efficacy class. DS composition showed 36 substances, 18 with no literature on male fertility and 18 showing positive effect on sperm parameters, thus considered potentially active ingredients (PAI). All DS were mixtures of ingredients, containing from 2 to 17 different substances. Fifteen supplements (65.2%) contained at least 1 ingredient without evidence of efficacy and 21 formulations had PAI dosed below mED. Some PAI were associated to the improvement of specific sperm parameters.

DS were usually blends of many substances, that are frequently employed at negligible dose or without any evidence of efficacy on male reproduction. Some ingredients have been demonstrated to be effective on specific sperm parameters by RCTs. We report a list of ingredients with potential efficacy on specific sperm parameters, aimed to allow a tailored use of DS.

In the light of our findings we rise three final considerations: i) the Italian market of DS for male infertility offers products with potential efficacy in the improvement of sperm parameters but also many with uncertain effect; ii) the actual literature is poor of well-designed studies on PAI investigating their mechanisms of action and effective dose in different pathological conditions; iii) based on current literature, our study can help in the choice of DS and PAI that are more likely to be effective on specific sperm alterations.

Our critical analysis suggests a rational strategy for a tailored use of DS in male infertility.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
RF02 | PMON309 Two Siblings with p.Arg92Trp Variant in NR5A1 Presenting with Testicular Disorder of Sexual Differentiation
Authors: Abisad Daniela Aguilar , Ballesteros Andrea Montano , Diaz Alejandro
Published: November 2022

Introduction

Disorders of sexual differentiation (DSD) are conditions of abnormal chromosomal development leading to atypical gonadal and anatomic sexual characteristics. We present the case of two siblings with 46, XX testicular DSD due to a mutation in the nuclear receptor subfamily 5 group A member 1 gene (NR5A1). While there is extensive literature about pathogenic variants of this gene, this is the first variant that has been shown to cause DSD in XX individuals, rather than XY.

Case presentation

This is the case of a sibling pair with male phenotype and history of hypospadias and hypogonadism in the setting of 46, XX karyotype. Sibling A presented with hypospadias at birth and hypergonadotropic hypogonadism in childhood, initially managed with testosterone. Testicular biopsy revealed bilateral germ cell aplasia with degeneration of seminiferous tubules. Sibling B presented at age 8 with precocious puberty. He had a history of hypospadias and cryptorchidism at birth. Gonadotropins and testosterone levels were prepubertal. Testicular biopsy showed only Sertoli cells. Their adrenal gland and thyroid function was normal, their bone age was advanced. FISH was negative for the SRY gene in both cases, no ovaries or mullerian structures found in pelvic imaging. DSD genetic panels were sent, resulting positive for the R92W variant in NR5A1. Neither of them have intellectual disability; however, sibling B has attention deficit and hyperactivity disorder (ADHD) and obsessive compulsive disorder (OCD). Family history is remarkable for two uncles that have been unable to conceive. Their mother had oligomenorrhea and difficulty conceiving, typical presentation of a female carrier of this mutation.

Discussion and conclusions

The majority of DSD in individuals with a 46, XX karyotype are due to congenital adrenal hyperplasia (CAH) secondary to 21-hydroxylase deficiency, however, mutations in NR5A1 have been among the most frequently identified genetic causes of gonadal development disorders and are associated with a wide phenotypic spectrum. NR5A1 is an essential transcription factor that regulates target genes crucial for normal reproductive physiology and endocrine function, controlling several steps of adrenal and gonadal development. While there is extensive literature about the phenotype caused by variants of NR5A1 in XY individuals, the recurrent heterozygous p.Arg92Trp NR5A1 variant R92W that our patients have, is the only one that has been shown to cause DSD in XX individuals, with variable degrees of testis development. It has previously been described, however, in women with primary ovarian insufficiency (POI). Morevariants of this gene will continue to arise, playing a pivotal role in reproductive endocrinology.

Presentation: Saturday, June 11, 2022 1:00 p.m. - 1:05 p.m., Saturday, June 11, 2022 1:00 p.m. - 1:05 p.m., Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
ODP144 Cushing Syndrome and Primary Hyperaldosteronism due to Synchronic Bilateral Adrenal Adenomas with Exclusive Production of Cortisol on the Right and Aldosterone on the Left
Authors: Rolim Marina Rocha , Mendonça Berenice Bilharinho , Ledesma Felipe , Charchar Helaine , Srougi Victor , Tanno Fabio , Chambo José Luis , Almeida Madson Queiroz , Latronico Ana Claudia , Fragoso Maria Candida Villares
Published: November 2022

Female patient, 37 years old, coming from China, with a history of resistant arterial hypertension since the age of 25 years old associated with spontaneous hypokalemia and classic ACTH-independent Cushing's syndrome. Radiological imaging exams showed bilateral adrenal nodules. Diagnosed primary hyperaldosteronism (Baseline aldosterone 103ng/dL (2.5-39.2), Renin 2.7uUI/mL (4.4 - 46.1), Aldo/renin ratio 38.1 and K 3.3) and confirmed laboratory hypercortisolism (Cortisol after 1mg of dexamethasone overnight 17.9mcg/Dl (< 1.8), salivary cortisol at midnight 0.466mcg/dL (<0.274), ACTH <2pg/mL, basal serum cortisol 19.4 (3.7–19.4). Abdominal CT identified bilateral adrenal nodules, one on the right and two on the left, with <10 HU, suggestive of adenomas. In the pandemic, we proposed the same surgical treatment for primary adrenocortical macronodular hyperplasia (total adrenalectomy of the largest gland and with greater uptake in PET-FDG and subtotal of the smallest and with less uptake). The patient was therefore submitted to total videolaparoscopic adrenalectomy on the left and subtotal on the right. The anatomopathological evaluation showed bilateral adenomas (Weiss 1) being the immunohistochemistry of the right adrenal nodule diffusely positive for CYP11B1 and negative for CYP11B2, inferring only cortisol production. In contrast, left adrenal nodules were positive only for CYP11B2, inferring exclusive aldosterone production. However, we also found bilateral subcapsular cell clusters with positivity for CYP11B2. This may infer a possibility of hyperaldosteronism recurrence. The patient developed secondary adrenal insufficiency postoperatively, requiring replacement with hydrocortisone, and cure of hyperaldosteronism with normalization of potassium and release of renin. Six months after the operation, there was no need for antihypertensive drugs. Currently, it is using only one class of antihypertensive, amlodipine 5mg a day, and it still needs to be replaced with hydrocortisone 20mg on waking up and 10mg at 2 pm with still undetectable serum cortisol concentrations, despite the ACTH being unblocked (17.3 pg /ml). Now we are working on genetic background of this case report.

Presentation: No date and time listed
RF34 | PMON254 Why Does Androgens Fail to Promote the Survival of the Wolffian Duct in the Absence of Wnt9b?
Authors: Crossen McKenna , Jia Shuai , Zhao Fei
Published: November 2022

The male reproductive tract development has been predominantly dependent upon androgen action, which is mainly mediated by the androgen receptor (AR). Under the influence of androgen actions, the primitive male reproductive tract, also known as the Wolffian duct, is first stabilized then differentiates into the epididymis, vas deference and seminal vesicle. Impairment of androgen signaling during fetal programming causes a spectrum of male reproductive disorders that manifest in adulthood. Despite the dominant roles of androgen action in the male reproductive tract development, it has been noticed that androgen's capability in promoting Wolffian duct survival and development is completely lost in the absence of Wnt9b. Wnt9b belongs to the WNT family of secreted proteins. It is specifically expressed in the Wolffian duct epithelium to regulate the surrounding mesenchyme. This project aims to investigate why the androgen action fails to maintain the male reproductive tract in the absence of Wnt9b. We first examined the expression pattern of Wnt9b during the window of sexual differentiation, which was not investigated in the previous publication. We found that during sexual differentiation, Wnt9b is still specifically expressed in the Wolffian duct epithelium in both XX and XY embryos. Androgen action exists in XY but not in the XX embryos during sexual differentiation. These observations indicate that Wnt9b does not lie downstream of the androgen signaling. We are in process of collecting Wnt9b+/+ and Wnt9b-/- male embryos to examine whether expression of AR and AR downstream genes is altered in the absence of Wnt9b. In addition, WNT9B can activate two intracellular pathways under the context of target cells: β-catenin dependent and independent pathways. Since Wolffian duct mesenchyme is the major target of epithelium-derived WNT9B, we next determined which pathways WNT9B acts through in the mesenchyme by specific deletion of β-catenin in the Wolffian duct mesenchyme. Interestingly, the loss of β-catenin in the mesenchyme led to the partial degeneration of Wolffian ducts at the caudal region and cystic formations at the cranial region, the similar phenotypes observed in mesenchymal AR knockout male embryos. Taken together, our study demonstrates that the local WNT9B/β-catenin pathway provides the permissive signaling for androgen actions in Wolffian duct development.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m., Monday, June 13, 2022 1:12 p.m. - 1:17 p.m.
RF34 | PMON276 The Effect of Gender Affirming Hormone Therapy (GAHT) on Non-Conventional Cardiovascular Risk Markers in Transgender Men: A Systematic Review
Authors: Moreira Allgayer Roberta Martins Costa , Gustavo da Silva Borba , Ramos Ramon Bossardi , Filho Ruy Silveira Moraes , Mara Spritzer Poli
Published: November 2022

The process of gender transitioning includes the employment of gender-affirming hormone therapy (GAHT) to attenuate gender dysphoria. Although considered relatively safe, there is still debate whether GAHT may increase the risk of cardiovascular disease (CVD) in transgender men (TM). Non-conventional subclinical cardiovascular (CV) risk markers, such as inflammation or coagulation markers, endothelial activity-related substances, and the use of ultrasound-assisted vascular function evaluation, may improve CV risk prediction in specific populations. Therefore, we conducted a systematic review to summarize the available findings regarding GAHT and its effects on these non-conventional cardiovascular risk markers in TM. Medline, LILACS, SCOPUS, and Embase databases were searched for cross-sectional, case-control, cohort, or randomized clinical trial studies published until July 2020. The selection criteria for the studies were as follows: ≥ 18 years old transgender male, no established CVD, using GAHT (androgens) for at least 2 months and compared with cis women or untreated transmen or before and during/after treatment for cohort studies. From 152 studies initially identified, 7 studies (3 cohorts and 4 case-control) were included in the systematic review. Some of these studies assessed different CV risk markers in the same group of participants. No changes were found in t-PA PAI-1, ICAMs, and PCR-us after 4 and 12 months of GAHT. Fibrinogen did not differ either in TM with or with no GAHT. Three studies evaluated flow-mediated-vasodilation (FMD) of the brachial artery. The only cohort study found no changes in FMD after 12 months of GAHT. One case-control study observed significant impaired FMD, and another one showed impaired FMD- after-nitroglycerin in transmen compared with cis women. One study reported increased homocysteine and higher carotid intima-media thickness (C-MIT) after 12 months of hormone use. Further, one study observed testosterone did not affect distensibility (DC) or compliance (CC) coefficient after 12 months, but the changes in fasting insulin levels were negatively associated with changes in the CC and DC. One study assessed the vascular function using brachial-ankle pulse wave velocity (baPWV) and observed a higher risk in TM with GAHT versus the untreated group. The results of the present systematic review suggest that GAHT may be associated with worse subclinical cardiovascular markers in TM. Further studies assessing long-term clinical follow-up of transgender men using GAHT could positively impact the prevention of CVD and management strategies for this population.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m., Monday, June 13, 2022 1:00 p.m. - 1:05 p.m.
ODP444 The Effect of Glucocorticoid Therapy on Cardio-Metabolic Risk in Patients with Multiple Sclerosis
Authors: Brovkina Snezhana , Dzherieva Irina , Volkova Natalya , Davidenko Ilya , Reshetnikov Igor , Komurdzhyants Maria , Tishchenko Yulia
Published: November 2022

Multiple sclerosis (MS) often requires usage of glucocorticoids (GC). Excess of GC may affect muscles, which increases mortality. This study evaluates the effect of GC therapy on muscles and cardiovascular disease (CVD) risk factors in patients with MS.

Methods

45 patients (55.5% women), 37.8 (20-45) years, were examined. 51% of patients received GC (methylprednisolone 3000-5000 mg, 3-5 days). MS duration was 5.9±1.2 years, the assessment by EDSS - 3.3±1.2. Muscle tissue was assessed by bioimpedance analysis, a chair-rising test, dynamometry. Glucose and lipid metabolism, blood pressure (BP), body mass index (BMI), waist circumference (WC),physical activity were evaluated. Statistical analysis was carried out using "Statistica 10. 0". Normal distribution was tested by the Shapiro–Wilks test, p>0. 05. А Mann-Whitney U-test, a t-test,χ2 test were used,p<0. 05.

Results

Patients with GC had significantly higher BMI (28.56±5. 02 vs 22.87±3.49 kg/m2), WC (98 (66-123) vs 79.9±5.27cm), lower muscle mass (36.4 (35-38) vs 40.75±2. 02 kg). A negative correlation between GC dose and a grip strength of the dominant hand among men was noted (r=-0.88, p<0. 0001). A normal WС was identified in 52% of patients. Comparing with others received GC, they had a lower level of low-density lipoproteins (1.35±0.39 vs 1.39(1-3,32) mmol/l), systolic BP (132.3±15.7 vs 141 (100-165)) and better handgrip strength (40 (25-47) vs 27±7.3 daN).

Conclusions

The negative effect of GC on cardiometabolic parameters in all patients with MS and muscle strength in men was confirmed. Metabolic healthier patients probably have a resistance to GC side effects, which may be determined genetically. It is promising for finding protective factors against GC.

Presentation: No date and time listed
OR07-2 Endocrine Response And Resistance Is Driven By Novel Transcriptional Co-Regulator Activity In Invasive Lobular Carcinoma Of The Breast
Authors: Costello James , Danis Etienne , Goodspeed Andrew , Mohammed Hisham , Sottnik Joseph , Sikora Matthew
Published: November 2022

Invasive lobular carcinoma of the breast (ILC) affects ∼40,000 US women annually and is a top ten most diagnosed cancer in women. ∼95% of ILC express estrogen receptor alpha (ER), but ILC is associated with poor long-term outcomes and anti-estrogen resistance, suggesting ER function is distinct in ILC cells. We reported that MDC1 (mediator of DNA damage checkpoint 1) is an ILC-specific ER co-regulator required for ER-driven proliferation, and that MDC1 knockdown dysregulates the ER transcriptome in ILC. Mechanisms of MDC1 co-regulator activity are unknown, but must be defined to understand endocrine response and resistance in ILC.

To understand MDC1 co-regulator functions, we profiled ER/MDC1 genomic binding (ie. "cistrome") using CUT&RUN in ER+ ILC cell line MDA MB 134VI (MM134). We identified ∼2500 binding sites each for ER and MDC1, but only ∼8% of sites were bound by both factors. However, using BETA to integrate cistrome and transcriptome data predicted that both ER and MDC1 activate estrogen-regulated genes with over half of direct ER target genes also direct MDC1 targets (p=9.8×10-375). Among shared ER: MDC1 direct target genes (n=760), 58% had MDC1 binding in the promoter region with ER bound at nearby enhancers, while only 15% were linked to shared ER: MDC1 binding sites, suggesting MDC1 primarily facilitates promoter access for ER-bound enhancers. Further, MDC1 knockdown reduced ER binding at enhancer sites at ER: MDC1 target genes, implicating MDC1 in regulating chromatin access.

Importantly, MDC1 does not have known enzymatic activity to directly regulate chromatin structure. We profiled MDC1-interacting proteins in ILC cells by RIME (Rapid immunoprecipitation mass spectrometry of endogenous proteins), and determined whether knockdown of putative ER: MDC1 partners disrupted ER function. This identified the SWI/SNF complex protein BRG1 (SMARCA4) as a critical ER: MDC1 partner. BRG1 was required for ILC cell growth, and BRG1 knockdown specifically suppressed ER regulation of MDC1-dependent ER target genes.

Taken together, our data suggest that in ILC cells, MDC1 facilitates ER access to and activity at target gene promoters. The BRG1 SWI/SNF complex is a putative ER: MDC1 partner and mediator of MDC1 co-regulator functions. Ongoing research is focused on understanding how MDC1 regulates ILC-specific ER activity by licensing access to ILC-specific target genes, and the role of the SWI/SNF complex in mediating genomic ER: MDC1 activity.

Presentation: Saturday, June 11, 2022 11:45 a.m. - 12:00 p.m.
OR26-2 mTOR Interacts with the Mineralocorticoid Receptor to Regulate its Activation by Ligand
Authors: Gomez-Sanchez Celso E , Gomez-Sanchez Elise P , Ali Yusuf
Published: November 2022

Background

Phosphorylation of the mineralocorticoid receptor (MR) at serine 843 inhibits its activity. mTOR phosphorylation of ULK1 prevents this MR phosphorylation.

Purpose

To study the relationship between mTOR and MR and its role in the modulation of aldosterone signaling.

Methods

Mouse cortical collecting duct M1 cells stably expressing the rat wild type MR or mutated MR (mu/S843A) and a Gaussia luciferase gene reporter under a hormone response promoter tyrosine aminotransferase (TAT) were incubated with mTOR activator and inhibitors in the presence and absence of aldosterone or corticosterone. Co-immunoprecipitation was performed to study protein interactions with the MR, and a phospho-serine antibody was used to demonstrate MR and ULK1 phosphorylation. Effects of mTOR knockdown in M1 cells by lentiviral transduction with CRISPR/gRNA for Raptor and Rictor were also studied.

Results

mTOR and the adaptor proteins of mTOR complex 1 and 2, Raptor and Rictor, respectively, co-precipitated with the MR of M1-rMR cells. Expression levels of these proteins were not changed by aldosterone. mTOR inhibitors significantly reduced ligand-induced MR reporter gene transactivation, however the mTOR activator MHY1485 had no effect. CRISPR/gRNA gene knockdown of Raptor or Rictor also inhibited MR activation stimulated by ligands, supporting a role of mTOR in maintaining the active MR. The mTOR inhibitor AZD8055 attenuated ligand-mediated MR gene transcription assessed by the reporter gene product and measurement of products of the endogenous MR target genes ACSBG1, LCN2 and Psca. AZD8055 also decreased phospho-ULK1 levels in a dose-dependent manner. We thus assumed that mTOR decreases ULK1 activity and prevents ULK1 phosphorylation of the MR at Serine (S843). Indeed, phosphorylation of the MR was increased in M1-rMR cells treated with AZD8055. However, AZD8055 still inhibited ligand-induced MR activity in cells carrying the mutated MR in which the Serine at 843 was replaced with an Alanine. These results suggest that mTOR has an additional role in MR activity unrelated to the specific phosphorylation at S843 by ULK1. This merits further investigation.

Presentation: Monday, June 13, 2022 11:15 a.m. - 11:30 a.m.
OR26-4 Glucocorticoid Receptor Expression and Signaling During Critical Illness, in Relation to the Duration of Illness and the Systemic Glucocorticoid Availability: A Prospective, Observational, Cross-Sectional Human and a Translational Mouse Study
Authors: Téblick Arno , Van Dyck Lisa , Van Aerde Nathalie , Vander Perre Sarah , Pauwels Lies , Derese Inge , Debaveye Yves , Wouters Pieter J , Vanhorebeek Ilse , Langouche Lies , Van den Berghe Greet
Published: November 2022

Critically ill patients are thought to develop maladaptive glucocorticoid-resistance which convinces many clinicians to administer stress doses of glucocorticoids to overcome this state of glucocorticoid-resistance. However, supportive data arises mainly from whole blood cells. It is currently not known if the observed changes in regulators and markers of glucocorticoid signaling and activity are also present in other cell and tissue types with a role in critical illness. We quantified regulators and markers of glucocorticoid signaling and activity in several cell and tissue types in critically ill humans and animals and in healthy controls. We found that throughout critical illness, glucocorticoid activity appeared suppressed in neutrophils, but upregulated in monocytes and skeletal muscle. Also in vital tissues GRα-signaling was altered in a tissue-specific, largely time-independent manner. Increasing systemic glucocorticoid availability increased glucocorticoid activity in adipose tissue, diaphragm and lung, whereas in immune cells and other tissues regulatory pathways counteracted. These data argue against glucocorticoid-treatable generalized glucocorticoid resistance and rather point towards an adaptive response in each specific cell or tissue type to optimally guide the beneficial actions of glucocorticoids to the tissues that need it while protecting collateral undesirable effects in tissue that are harmed by elevated systemic glucocorticoid availability.

Presentation: Monday, June 13, 2022 11:45 a.m. - 12:00 p.m.
OR26-6 | LBMON298 The Role Of Nuclear Receptor Coregulator Grip1 In Homeostatic Programming Of Macrophages
Authors: Dacic Marija , Deochand Dinesh , Deochand Yurii , Rogatsky Inez
Published: November 2022

Aberrant macrophage function is implicated in acute and chronic inflammatory settings, and central to diseases such as obesity, atherosclerosis and cancer. However, the transcriptional and epigenomic programs driving macrophage polarization toward distinct phenotypic states are not well understood. Macrophage phenotypes exist on a spectrum, with pro-inflammatory (M1-like) and homeostatic, tissue repair (M2-like) states viewed as polar opposites and relatively stable. The main driver of the M2-like program is a type-2 cytokine, IL4 which signals to STAT6 followed by the Krüppel-like factor 4 (KLF4), a master regulator for numerous M2-specific genes. Reportedly, glucocorticoid (GC) hormones also promote the M2-like state, although the extent to which the phenotypes elicited by such disparate signals overlap is unknown. Interestingly, we discovered that GRIP1, a well-known cofactor for nuclear receptors including GR, serves as a KLF4 coactivator, suggesting that GRIP1 could mediate transcriptional and phenotypic convergence of the IL4- and GC-driven pathways. Gene expression profiling on differentially polarized macrophages revealed a highly significant overlap between M2IL4 and M2GC transcriptomes which was mirrored by the H3K27ac de novo enhancer landscape. Notably, loss of GRIP1 blunted both up-regulation of M2- and downregulation of M1-associated genes irrespective of the signal driving M2 polarization. At the cellular level, polarization with either IL4 or GCs enhanced macrophage phagocytosis relative to non-polarized M0, and did so in a GRIP1-dependent manner. Finally, in the dextran sulphate sodium (DSS)-induced colitis model of intestinal inflammation and tissue damage, GRIP1-cKO mice lacking GRIP1 in myeloid cells, including macrophages, displayed more severe destruction of the colonic mucosa and greater immune cell infiltration into the mucosa and submucosa. Moreover, in total colon RNA, the abundance of many characteristic M2 transcripts was reduced in the cKO mice. Surprisingly, the expression of proinflammatory mediators such as TNF, IL1b and IL23 was unaffected by GRIP1 deletion, suggesting that the more severe disease in the cKO stems primarily from the failure of M2-like resolving macrophage to efficiently mitigate tissue destruction and repair the damage. Together, these findings point to GRIP1 as a crucial regulator of macrophage polarization to the homeostatic state in vitro and in vivo.

Presentation: Monday, June 13, 2022 12:15 p.m. - 12:30 p.m., Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON285 Hypokalemic Periodic Paralysis: A Rare Presentation of Primary Hyperaldosteronism
Authors: Elsheikh Marwa , Robertson Anna-Belle , Lavine Ross , Abdelhadi Mahmoud , Rastogi Natasha
Published: November 2022

Background

Primary hyperaldosteronism (PHA) is caused by excessive production of aldosterone from the adrenal glands. Aldosterone levels are usually independent of renin, angiotensin II or sodium status, and often result in hypertension (HTN), hypokalemia and metabolic alkalosis. The most common presentation of this condition is with asymptomatic mild to severe or refractory HTN. This case highlights a patient presenting with hypokalemic periodic paralysis as the first sign of PHA.

Clinical Case

A 35 year old previously healthy Chinese male, presented with sudden onset bilateral lower extremity weakness for 1 day. This was associated with mild transient lower back pain and fatigue. He had no history of trauma, recent travel, exposure to animal or tick bites. He reported no recent illness or vaccination. His family history was negative for hypertension and various endocrinopathies. The patient was not on any medications and did not consume licorice.

His exam was significant for a blood pressure (BP) of 217/136 mmHg, with lower limb evaluation revealing power of 3/5, normal reflexes and a positive heel to shin test. He demonstrated an unsteady gait but Romberg's sign was negative. Upper limb exam was normal. Laboratory findings were remarkable for potassium 1.9 mmol/L, bicarbonate (CO2) 37 mmol/L, normal magnesium (Mg), normal TSH and blood gas showing metabolic alkalosis. Subsequent testing showed renin 0.35 ng/ml/hr, aldosterone 23 ng/dl and aldosterone/pra ratio 66. Saline infusion test confirmed the diagnosis of PHA. Other endocrinologic tests were normal. Electrocardiogram showed prolonged QT interval and U waves. Renal ultrasound was negative for renal artery stenosis and CT abdomen/pelvis showed a 2.4 cm left adrenal adenoma.

The patient was admitted to the intensive care unit for 24 hours, where he received intravenous potassium replacement and was started on spironolactone and other antihypertensive medications, which he was discharged on, along with oral potassium. His symptoms resolved after potassium levels normalized and he subsequently had a left adrenalectomy.

Conclusion

With PHA being the most common endocrinological cause of HTN, clinicians should recognize indicators of underlying PHA even in rare presentations such as this.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON295 Investigating the ileal FXR/SHP axis in regulating lipid absorption and enterohepatic recirculation of bile acids
Authors: Nguyen James , Anakk Sayeepriyadarshini
Published: November 2022

Lipids are biological macromolecules essential for energy storage, signaling pathways, and cell structure. Majority of these molecules come from our diet where they are digested in the small intestine through their interaction with bile acids. Lipids break down into small molecules and are brought to the intestinal brush border to be absorbed passively by diffusion or by lipid transporters. Interestingly, two essential nuclear receptors that regulate both bile acid and lipid absorption and transportation are farnesnoid X receptor (FXR) and small heterodimer partner (SHP). Many studies have shown that the linear FXR/SHP axis controls these lipid metabolic processes, however, there are evidence indicating FXR-dependent and -independent pathways of SHP. In order to ascertain the importance of FXR and SHP in intestinal lipid metabolism, we utilized the following three mice models, intestine-specific Fxr (IFxrKO), intestine-specific Shp (IShpKO), and intestine-specific Fxr and Shp (IDKO) knockout models and challenged them with high fat diet. We are assessing the intestinal absorption and transportation of triglycerides by measuring the concentration of triglyceride on circulation, liver, and intestine and the molecular level the protein expression of different transporters. Parallelly, we will examine the bile acid pool by measuring bile acid levels from the intestine, liver, gall bladder, and serum to test if changes in bile acid circulation observed in these knockout models show different contribution towards lipid/triglyceride absorption. In addition, we will inject poloxamer-407 in these mice groups to assess their chylomicron secretion rate. Finally, we will examine the histology of the small intestine by performing H&E staining to examine the morphology and perform PAS/Alcian blue staining to examine mucus production.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.