bluuebunny/crossref_metadata_2025 huggingface dataset:
5178 TNFα-dependent altered miRNA expression regulates endometrial stromal cell proliferation
Authors: Chowdhury I , Banerjee S , Xu W , Driss A , Nezhat C , Sidell N , Taylor R N , Thompson W E
Published: October 2024

Disclosure: I. Chowdhury: None. S. Banerjee: None. W. Xu: None. A. Driss: None. C. Nezhat: None. N. Sidell: None. R.N. Taylor: None. W.E. Thompson: None.

Abstract: Aberrant upregulation of cytokines, including tumor necrosis factor α (TNFα) and activation of nuclear factor κB (NF-κB) with disbalance of microRNAs (miRNAs) are often found in endometrium of endometriosis subjects, which causes directly or indirectly pelvic inflammation, pain, and infertility, Although, the pathophysiology of endometriosis is not clearly established yet, inflammation is an essential feature. MicroRNAs are emerging as potential regulatory factors in cellular processes such as cell growth, survival, migration, and invasion. In the current study, we examined the direct effects of exogenous recombinant TNFα dependent regulation of miRNAs involved in proliferation in primary cultures of normal endometrial stromal cells (NESC) and compared with the untreated cells derived from endometriosis-ESC (EESC). NanoString nCounter-based assays and quantitative real-time polymerase chain reaction (RT-PCR) were used to identify and validate differentially expressed miRNAs linked to cell proliferation in EESC compared to NESC and TNFα treated NESCs. The expression level of miRNAs linked to cell cycle was downregulated significantly in EESCs compared to NESCs. Interestingly, a dose and time-dependent exogenous TNFα treatment in NESCs showed a similar cell cycle regulator miRNAs expression pattern with EESCs. In further studies, protein analysis by Western Blots of selective cell cycle regulators such as PCNA, Ki67, CDK2, CDK4, CDK6, p21, and p27, and cell proliferation assay demonstrated an inverse correlation between cell cycle regulators and miRNA expression pattern both in EESC and TNFα treated NESCs. These findings suggest that TNFα dependent dysregulation of miRNAs in EESCs may promote proliferation of endometriosis. Further research is needed to investigate the detailed mechanisms by which TNFα regulates miRNA expression and determine these findings' potential therapeutic implications. Acknowledgments: This study was supported in part by National Institutes of Health Grants 1SC3 GM113751, 1SC1 GM130544, U01 HD66439, 1R01HD057235, U54 CA118948, HD41749, S21MD000101, and G12-RR03034. This investigation was conducted in a facility constructed with support from Research Facilities Improvement Grant #C06 RR18386 from NIH/NCRR.

Presentation: 6/2/2024
12545 Transgender Healthcare Online Curriculum: A Partnership Between An Academic Medical Center And Corrections Facility
Authors: Lamos Elizabeth , Malek Rana
Published: October 2024

Disclosure: E. Lamos: None. R. Malek: None.

Introduction: The World Professional Association for Transgender Health Standards of Care (WPATH SOC-8) states that all recommendations be applied equally to individuals living in institutions​, including those incarcerated. Studies suggest correctional staff have little training in transgender care.​ To demonstrate a partnership between an academic medical center and a corrections facility and increase knowledge and comfort in transgender healthcare, academic faculty developed an online curriculum as an educational opportunity for state corrections facility staff and clinicians. Methods: There were 6 modules: Overview of Transgender Competent Care, Health Vulnerabilities, Creating an Inclusive Environment, Feminizing and Masculinizing Hormone Treatment, and Preventative Care of the Transgender Individual (Image). The Short Course (first 3 modules) was for staff or clinicians not providing hormone-affirming care and the Full Course (all 6 modules) for clinicians who provide hormone-affirming care. Each module included embedded knowledge assessments and post-module action items. An anonymous pre and post curriculum survey assessed module effectiveness. The project was considered non-human subject research by the Institutional Review Board. Result analysis used the Fisher Exact Test and unpaired t test. Results: 44 corrections staff and clinicians participated (2022-2023) with a 32% post survey response rate. 57% indicated zero prior hours of transgender education with 59% feeling somewhat or extremely comfortable with transgender healthcare. Most were interested in learning about transgender healthcare (Table 1). Participants had a statistical improvement in knowledge and their access to resources were good or excellent after completion (p <0.05, both) (Table 2). Also, for the Short Course, comfort in recognizing barriers to care significantly improved (p<0.05), 100% agreed both with the importance of what they were asked to learn and that knowledge and skills increased. Domains of comfort with transgender healthcare did not statistically change for participants of the Full Course. Participants positively reflected on curriculum content and bias discussion. Full Course respondents requested more therapeutics information. Inclusive interactions were the most frequent practice improvement strategy. Conclusion: This curriculum increased knowledge and improved resources for those providing healthcare to incarcerated transgender patients. The curriculum was inclusive of many diverse types of learner and allowed learners to choose the course that aligned with their clinical role. It meets WPATH SOC-8 recommendations that corrections staff and clinicians receive training in transgender care.​ It highlights the collaboration between an academic medical center and corrections facility, meeting the academic mission to serve marginalized groups.

Presentation: 6/1/2024
12209 Validation Of A Novel Placenta Explant Protocol For Studying Circadian Rhythms In The Mouse Placenta In Early, Mid And Late Pregnancy
Authors: Lee Jessica S , Yaw Alexandra M , Hoffmann Hanne Mette
Published: October 2024

Disclosure: J.S. Lee: None. A.M. Yaw: None. H.M. Hoffmann: None.

Introduction: The placenta serves as an interface facilitating the exchange of essential nutrients and oxygen between the mother and fetus. Preeclampsia, a complication of pregnancy driven by placental dysfunction, is one of the leading causes of maternal, fetal, and neonatal morbidity and mortality. While deregulation of the genes driving circadian rhythm has been associated with preeclampsia, the role of circadian rhythms in the placenta largely remains unknown. To address this gap of knowledge, our goal was to establish and validate an organotypic ex vivo preparation to study placenta circadian rhythms at gestation day (GD) 10, GD14, and GD18 in the mouse. Methods: To establish an organotypic placenta culture GD10, GD14, and GD18 placental explants, we sectioned the placenta into 350μm sections using a vibratome. The explants were prepared from the validated circadian Per2::Luciferase reporter mouse, allowing automated circadian rhythms recording of the explants in a LumiCycle. We compared explant success rate and rhythm quality in 2x2mm section vs whole slice explants at GD14. Using hematoxylin and eosin (H&E) staining we validated the explants to be decidua which is strictly of maternal origin, labyrinth which is strictly of fetal origin, and the junctional zone which consists of binucleated cells fused of maternal and fetal cells. Results: Through H&E staining, we confirmed the decidua is enriched with neutrophils, the junctional zone contains spongiotrophoblasts and glycogen trophoblasts, and the labyrinth has red blood cells. Both explant types were enriched with >80% of cells from the targeted region. We next established a protocol to record Per2::Luciferase rhythms for <6 days. The fragility of the GD10 placentas required the whole explant protocol, whereas the GD18 2x2mm sectioned explants produced strong rhythms with a high success rate. At GD14, the whole explants resulted in stronger rhythms with higher amplitude and higher goodness of fit compared to the 2x2mm explants. Conclusion: We found that depending on GD recording success differs. Specifically, the whole placental explant provides a more robust and reliable model for studying circadian rhythms in GD10 and GD14, whereas 2x2mm explants are successful at GD18. This novel placental preparation protocol will be valuable in studying disease- and drug-induced changes to circadian rhythms in a placenta layer-specific manner in the mouse.

Presentation: 6/1/2024
7542 Androgen Receptor Oligomerization State Allows Cells to Exhibit Non-Linear Dose Responses to Androgens Resulting in Distinct Biological Outputs
Authors: Safi R , Wardell S E , Watkinson P , Qin X , Lee M , Park S , Krebs T , Dolan E , Tsuji T , Nayak S , Khater M , Newlin M , Kirkland M , Xie Y , Long H , Fink E , Fanning S W , Runyon S , Brown M , Xu S , Owzar K , Norris J D , McDonnell D P
Published: October 2024

Disclosure: R. Safi: None. S.E. Wardell: None. P. Watkinson: None. X. Qin: None. M. Lee: None. S. Park: None. T. Krebs: None. E. Dolan: None. T. Tsuji: None. S. Nayak: None. M. Khater: None. M. Newlin: None. M. Kirkland: None. Y. Xie: None. H. Long: None. E. Fink: None. S.W. Fanning: None. S. Runyon: None. M. Brown: None. S. Xu: None. K. Owzar: None. J.D. Norris: None. D.P. McDonnell: None.

An unanswered fundamental question in androgen action is how cells recognize and respond in a non-linear fashion to different levels of androgens to elicit distinct biological outputs such as occurs in prostate cancer (PCa) cells. Indeed, most PCa express the androgen receptor (AR), and tumor growth and progression are facilitated by exceedingly low levels of systemic or intratumorally produced androgens. Paradoxically, high dose androgens have been used to treat patients with late-stage metastatic PCa. We have determined that androgen dose regulates the relative cellular abundance of AR monomers and dimers in cells and that the biological output of each oligomeric form of the receptor is different. Using in vitro systems that model exposure from castrate (low dose; LD) to eugonadal levels and above (high dose; HD) of androgens, we have determined that the global changes in gene expression are substantially different, with the changes induced by LD androgens (monomeric AR) being associated with cell proliferation, and HD androgens (dimeric AR) inducing a differentiated phenotype. A similar distinction in response to androgen levels was observed in vivo. Using a chemical biology strategy, we identified ligands which freeze AR in a monomeric form and demonstrate that these compounds faithfully recapitulate the LD biology exhibited by physiological AR agonists. Mechanistically, we determined that monomeric AR facilitates a non-genomic activation of the mTOR signaling pathway to drive proliferation, while the dimer suppresses c-MYC expression, inhibit proliferation, and drive a transcriptional program associated with a differentiated phenotype. These findings highlight the inherent liabilities in current approaches used to inhibit AR-action in prostate cancer and are instructive as to strategies that can be used to develop new therapeutics for this disease and other androgenopathies.

Presentation: 6/3/2024
6609 Central Serous Chorioretinopathy in the Eyes of the Beholder of Testosterone Depot Injection
Authors: Ni Huajing , Kankanamge Denuka , Jabbour James , Preda Veronica Angela
Published: October 2024

Disclosure: H. Ni: None. D. Kankanamge: None. J. Jabbour: None. V.A. Preda: None.

Background: Central serous chorioretinopathy (CSC) is a characterised by subretinal fluid accumulation causing detachment of the neurosensory retina and visual disturbance[1]. It has been postulated that increased focal choriocapillaris permeability results in leakage of interstial fluid into the subretinal space[1]. CSC is a rare side effect of testosterone therapy, the exact mechanism is unclear. Case : We present a 32-year-old male gym manager who has been self-administering anabolic steroids with testsoterone ethanoate IM injections every 14 days for body building. He reported left blurred vision for 8 weeks. He was normotensive and reported taking doxycycline and azithromycin for mycoplasma genitalia. There was no history of glucocorticoid exposure, smoking, alcohol use, gastoesophageal reflux disease, hypertension, shift work, sleep apnoea, nor other established risk factors for CSC. The visual acuity was OD 6/6 and OS 6/9 with intraocular pressures of 12mmHg on the right and 15mmg on the left. Fundoscopy revealed a left serous macular elevation involving the left fovea. A macular OCT confirmed the left acute central serous chorioretinopathy and right retinal pigment epithelial changes. The latter were suggestive of right CSC in the past. Discussion: Androgens have been associated with CSC in the literature. It has been hypothesised that exogenous testosterone may cause increased choriodal blood flow and permeability, promote atheroscleorsis, cause sodium retention and potentially interact with retinal pigment epithelium (RPE) via androgen receptors4. Intramuscular testosterone therapy causes fluctuation in plasma testosterone concentration (which is thought to be more relevant than absolute value of testosterone). Most cases (60%) of CSC resolve spontaneously, and appear to coincide with cessation of testosterone therapy5. Conclusion: Clinicans need to be alert to the use of testosterone and other anabolic steroids, prescribed or self administered, particularly in their patients with CSC or who have other risk factors for CSC, such as corticosteroids, smoking and hypertension. References: 1.Kaye R, Chandra S, Sheth J, Boon CJF, Sivaprasad S, Lotery A. Central serous chorioretinopathy: an update on risk factors, pathophysiology and imaging modalities. Prog Re- tin Eye Res. 2020 Nov; 79:100865. 2.Kitzmann AS, Pulido JS, Diehl NN, et al. The incidence of central serous chorioretinopathy in Olmsted County, Minnesota, 1980–2002. Ophthalmology 2008; 115:169–173. doi: 10.1016/j.ophtha.2007.02.032. 3.Ahad MA, Chua CN, Evans NM. Central serous chorioretinopathy associated with testosterone therapy. Eye. 2006 Apr;20(4):503– 5.4.Nudleman E, Witmer MT, Kiss S, Williams GA, Wolfe JD. Central serous chorioretinopathy in patients receiving exogenous testosterone therapy. Retina. 2014;34(10):2128–32.

Presentation: 6/2/2024
8808 Characterization and pre-clinical study on a novel hormonal compound with potent analgesic and anti-inflammatory properties
Authors: Antakly T , Brox A , Menezes J , Zingg H H
Published: October 2024

Disclosure: T. Antakly: Other; Self; Theriac Biomedical. A. Brox: Owner/Co-Owner; Self; Theriac Biomedical. J. Menezes: Advisory Board Member; Self; Theriac Biomedical. H.H. Zingg: None.

Hormones and naturally occurring compounds with therapeutic benefits have provided the basis of some of the most celebrated drugs e.g. Taxol (extracted from the Pacific Hew tree) and Premarin, a hormone replacement therapy (extracted from horse urine). Our team has discovered and chemically identified a bioactive compound (called “HIP-2”) found in folk medicinal admixtures. It has unique analgesic and anti-inflammatory properties. The chemical structure of HIP-2 consists of small fatty acid molecules which bind and activate the PPAR-gamma transcription factor, and furthermore they have HDAC inhibitory properties. A synthetic replica of the bioactive compounds was obtained and administered orally or by injection in a single dosage form. Using animal models for pain and inflammation, HIP-2 has demonstrated an outstanding efficacy and safety profile in animals (rats, mice) rendered experimentally inflamed (e.g. carrageenan injection) or osteoarthritic (surgery or iodoacetate injection). Pain and inflammation have decreased (by up to 50%) after HIP-2 administration. Inflammatory cytokines biomarkers were also reduced by about 50% as compared to baseline or to the positive controls (classical NSAID’s). HIP-2, with an outstanding safety and efficacy profile, offers an advantage over currently available analgesic and anti-inflammatory drugs, which have significant draw-backs due to limited efficacy and significant side effects such as gastrointestinal bleeding/ulcers/perforation and cardiac toxicity (Advil, Celebrex), osteoporosis and immunosuppression (corticosteroids), or addiction (opioids). As a potential solution to these problems, HIP-2 is likely to be superior both at the safety and efficacy levels. The HIP-2 IMP was cleared by FDA and is entering Phase 2 clinical trials.

Presentation: 6/3/2024
7005 Characterization of Clinical Candidate Miricorilant in Preclinical Models for NASH
Authors: Moll Tijmen J A , Viho Eva M G , Gentenaar Max , Hunt Hazel , Meijer Onno Cornelis , Kroon Jan
Published: October 2024

Disclosure: T.J. Moll: None. E.M. Viho: None. M. Gentenaar: None. H. Hunt: Employee; Self; Corcept Therapeutics. O.C. Meijer: Grant Recipient; Self; Corcept Therapeutics. J. Kroon: Research Investigator; Self; Corcept Therapeutics.

Background and aim: Miricorilant (CORT118335) is a selective glucocorticoid receptor (GR) modulator that is under clinical development for the treatment of non-alcohol steatohepatitis (NASH). It was previously shown that miricorilant treatment effectively lowers hepatic lipid content in mouse models of NASH. The aim of this study is to further characterize the effects of miricorilant in the mouse liver and in human liver tissue. Methods: In mouse liver tissue, we investigated the agonistic and antagonistic properties of miricorilant on GR activity. Eight-week-old male C57BL/6J mice were fed with a control diet or miricorilant-supplemented diet (resulting in an estimated dose of 60 mg/kg/day) for 6 days and 1.5 mg/kg corticosterone was injected to monitor the effects of miricorilant on GR-responsive gene expression. We next accessed the time-dependency of the lipid-lowering effects of miricorilant in the mouse liver. Mice were fed with a high-fat, high-fructose diet (HFHFD) for 3 weeks before treatment with 60 mg/kg/day miricorilant for one, three or six days by oral gavage, and triglyceride content in the liver was measured at endpoint. In human liver tissue, we investigated the effects of miricorilant on precision-cut liver slices (PCLS) obtained from two independent donors with pre-existing steatosis. PCLS were cultured for a total of 168 hours in the presence of 100 nM hydrocortisone, and with or without an exogenous lipid challenge. PCLS were treated with 200-600 nM miricorilant for 96 hours before triglyceride content was measured. RNA-sequencing was performed on PCLS after 24 hours of 200-400 nM miricorilant exposure. Results: In the mouse liver, miricorilant exhibited a unique combination of agonistic and antagonistic properties on GR-responsive gene expression. Miricorilant effectively lowered HFHFD-induced lipid accumulation in the mouse liver as soon as three days after treatment. In human PCLS, miricorilant treatment lowered triglyceride content in both donors. RNA-sequencing revealed respectively 40 and 128 differentially expressed genes after 200 and 400 nM miricorilant treatment. Pathway analysis revealed very-low-density lipoprotein (VLDL) particle remodeling as a significant enriched pathway, providing a possible explanation for the lowered triglyceride content upon miricorilant treatment. Conclusion: Miricorilant rapidly and effectively lowers hepatic lipid content in the mouse liver and in human PCLS, possibly via modulation of VLDL remodeling.

Presentation: 6/2/2024
8672 Disrupted Glucocorticoid Receptor Mediated Signalling Causes a Primary Cilia Defect in the Fetal Mouse Renal Tubule
Authors: Lao J , Short K L , Ng J , Cottle D L , Cole T J
Published: October 2024

Disclosure: J. Lao: None. K.L. Short: None. J. Ng: None. D.L. Cottle: None. T.J. Cole: None.

Primary cilia are microtubule-based organelles that protrude from cell membranes to mediate diverse developmental signalling pathways and senses extracellular stimuli to maintain tissue homeostasis. We show that glucocorticoid (GC) signalling via the glucocorticoid receptor (GR) is a regulator of normal primary cilia formation in kidney renal tubules. RNA sequencing of whole E18.5 fetal kidney from mice with global deletion of the GR (GR-null) compared to littermate wild-type controls identified significantly reduced mRNA levels of key ciliogenesis-related genes. Further analysis of whole fetal kidney RNA by real-time-qPCR confirmed reduced mRNA levels for the cilia-associated proteins Ccp110 (fold -2.18), Cep97 (fold -1.78), Cep290 (fold -2.91), Kif3a (fold -1.82) and Rpgr (fold -1.90). Confocal microscopy revealed abnormal, stunted primary cilia in renal proximal tubules, collecting ducts and podocytes in GR-null or in renal tubule conditional GR-deleted mice, created using the HoxB7 Cre-driver allele. Primary cilia length was significantly decreased in kidney proximal tubule cells in GR-null mice (5.01 ± 0.11μm) compared with wildtype controls (6.20 ± 0.15μm). Furthermore, scanning electron microscopy of E18.5 GR-null kidney further confirmed aberrant primary cilia morphology in the proximal renal tubules and the absence of an apical brush border. Activation of GR signalling with the synthetic GC dexamethasone in cultured mouse IMCD3 kidney tubule cells significantly increased primary cilia length (2.89 ± 0.04μm) compared to vehicle controls (2.46 ± 0.28μm), an effect blocked by the GR antagonist RU486 (vehicle + RU486: 2.06 ± 0.02μm, dexamethasone + RU486: 2.00 ± 0.02μm). Aurora kinase A (AURKA) is a known regulator of primary cilia formation via the AKT cell signalling pathway and distinct proteins at different stages of the mitotic cell cycle. AURKA protein levels were downregulated in GR-null kidney and in IMCD3 cells which suggest GR regulates AURKA to control the assembly and disassembly of primary cilia. Together, these results demonstrate that GC signalling via the GR is required for normal primary ciliogenesis in the developing renal tubule and suggests that synthetic GR agonists may provide a novel therapeutic option for human ciliopathies such as those observed in forms of polycystic kidney disease.

Presentation: 6/2/2024
7340 Elucidating the Role of Steroid Hormone Receptors in Anaplastic Thyroid Carcinoma and Pancreatic Ductal Carcinoma Models
Authors: Stockert O M , Lange C A
Published: October 2024

Disclosure: O.M. Stockert: None. C.A. Lange: None.

Steroid hormones play important roles in modulating transcription mechanisms in a variety of cancers, including breast cancer, prostate cancer, and ovarian cancer. These interactions are highly context-dependent and regulated by diverse ligands, cofactors, and dynamic post-translational modifications that occur in response to the activation of stress-sensing and oncogenic signaling pathways. Numerous other cancers are associated with elevated steroid hormones, steroid hormone receptors, or both, but have not been studied to explore the possibility of leveraging steroid or steroid receptor (SR)-modulated transcriptional regulation to combat cancerous growth. Two of these cancers are pancreatic ductal adenocarcinoma (PDAC) and anaplastic thyroid carcinoma (ATC). PDAC is an aggressive cancer with a very poor 5-year survival of 8-11%. It is also rapidly growing in incidence and has been shown to be associated with other increasing health conditions, including diabetes and obesity. PDAC is associated with metabolic reprogramming and elevated Progesterone Receptor (PR), Glucocorticoid Receptor (GR), and Mineralocorticoid Receptor (MR) expression. ATC is another aggressive cancer with less than 10% 6-month survival from diagnosis. Like PDAC, ATC has a rapidly increasing incidence and is associated with obesity and high-fat diets. ATC is also associated with elevated cholesterol biosynthesis and elevated GR expression. Herein, we probed public data sets and screened a panel of PDAC and ATC models to demonstrate the expression of mixed SRs. In this study, we take an initial exploration of the hypothesis that related SRs (such as PR, GR, and/or MR) contribute to PDAC and ATC progression and that post-translational modifications, ligand binding, and/or selected cofactors modulate SR actions in the context of activated signaling pathways associated with these aggressive cancers. This study gives us a preliminary understanding of the potential role of steroid hormones and their receptors in the aggressive biology of two deadly and increasingly prevalent endocrine cancers in aging and obese populations.

Presentation: 6/3/2024
12539 Estrogen Type In Contraceptives Matters: Effects On Spatial Memory And Anxiety-Like Behaviors In A Female Rat Model
Authors: Hegwood A P , Kujawa S , Prakapenka A V
Published: October 2024

Disclosure: A.P. Hegwood: None. S. Kujawa: None. A.V. Prakapenka: None.

According to U.S. Centers for Disease Control & Prevention, ∼12% of the U.S. population uses combined oral contraceptives (COCs). Behavioral side effects are common factors in COC discontinuation. Management of COC-related side effects primarily centers around altering the synthetic ethinyl estradiol (EE) dose, the predominant estrogen in COCs. Estradiol valerate (EV) is a recently introduced natural estrogen alternative to EE. EE-containing COCs, such as EE/dienogest (DNG), were found to decrease free steroid hormone serum levels compared to EV-containing COCs, such as EV/DNG. In women and in female rodent models, steroid hormones (e.g., 17-ß estradiol (E2), testosterone) dose-dependently modulate spatial memory and anxiety-related behaviors. We hypothesized that in a rat model of hormonal contraceptive use, EE/DNG differentially modulates spatial memory and anxiety-like behaviors compared to EV/DNG, corresponding to circulating steroid hormone levels. Thirty-six young-adult (3-4-months-old) female rats were randomly assigned to EE/DNG (0.13µg EE and 8.6µg DNG per day), EV/DNG (8.6µg EV and 8.6µg DNG per day), or vehicle treatment (propylene glycol) continuously administered by subcutaneous osmotic pump. At four weeks of treatment, rats were tested on the delayed spontaneous alternation (dSA) task to evaluate spatial memory and the elevated plus maze (EPM) to evaluate anxiety-like behavior. Following EPM, serum was collected and analyzed for E2, progesterone, androstenedione, and testosterone levels using liquid chromatography-mass spectrometry and corticosterone levels using radioimmunoassay. Spontaneous alternation behaviors were similar across all three groups on the dSA. However, EE/DNG increased use of an egocentric navigational strategy, the use of consecutive turns, on the dSA compared to vehicle and EV/DNG. Egocentric strategy is likely mediated by the striatal memory system, engagement of which is elevated with stress and anxiety. Of note, EE/DNG decreased time spent in the open arms of the EPM compared to vehicle and EV/DNG, indicating increased anxiety-like behaviors with EE/DNG. Both EE/DNG and EV/DNG decreased androstenedione, testosterone, and E2 circulating levels compared to vehicle. Steroid hormone serum levels did not differ between EE/DNG and EV/DNG; 58% of EE/DNG serum samples were undetermined/below threshold for E2 levels, limiting statistical analysis. Altogether, we found that, compared to vehicle and the natural EV, the synthetic EE component in hormonal contraceptives elevates anxiety-like behaviors in female rats, an effect that corresponds to increased use of an egocentric navigational strategy on a spatial memory task. Our findings support the notion that estrogen type matters for behavioral outcomes associated with contraceptive use, identifying estrogen type as a possible tool for management of COC-related side effects.

Presentation: 6/1/2024
6891 Exclusive Normetanephrine Secreting Pheochromocytoma In A Young Male
Authors: Askar Esraa , Moran Heberth , Yuksel Simge , Bleich David
Published: October 2024

Disclosure: E. Askar: None. H. Moran: None.

IntroductionPheochromocytomas and paragangliomas arise from chromaffin cells and produce excessive catecholamines. Hypertension is their primary clinical symptom. they differ in their catecholamine secretory phenotype; paragangliomas typically secrete norepinephrine, while pheochromocytomas typically secrete epinephrine. Here, we present a case 21-year-old male with pheochromocytoma with elevated norepinephrine but normal epinephrine levels.Case presentationA 21 -year-old male with no past medical history presented with episodes of palpitations, sweating, and elevated blood pressure. He also reported abdominal pain. On admission, his blood pressure was 143/92 mmHg, pulse was 71/min, respiration was 16/min, and oxygen saturation was 100% in room air. The rest of physical exam was not significant. His electrolyte panel was normal. Computed tomography (CT) abdomen and pelvis with contrast was performed and showed a right adrenal mass measuring 4.9 x 4.0 x 6.5 cm which is predominantly hypoattenuating (approximately 30 Hounsfield units) with thick nodular rim of enhancement. Hormonal workup showed normal serum testosterone, DHEA-S, AM cortisol, ACTH, aldosterone, plasma metanephrine, and urine metanephrine, but profoundly high plasma normetanephrine 3641.6 (0.0-210.1) pg/mL. His 24-hour urinary normetanephrine was elevated at 14874 (80-440) ug/24hr, confirming a pheochromocytoma. The high norepinephrine level in the setting of normal epinephrine raised suspicion for Von Hippel Lindau (VHL). He was started on alpha and beta blockage and had laparoscopic removal of the right adrenal gland. The pathology report confirmed the diagnosis of pheochromocytoma confined to right adrenal. The patient was then referred for genetic testing. Conclusion: Pheochromocytoma can be a part of familial disorders, although being sporadic in nature most of the time. As demonstrated in our case, the hallmark of pheochromocytoma associated with VHL is nearly exclusive normetanephrine secretion and a high ratio of normetanephrine to metanephrine. Confirmation and testing of genes aid in proper monitoring and follow-up. The preferred course of treatment is radical resection.

Presentation: 6/2/2024
8236 Hepatocyte Vitamin D Receptor (VDR) Regulates Liver Regeneration
Authors: Gunton J E
Published: October 2024

Disclosure: J.E. Gunton: None.

Harendran Elangovan [1], Rebecca A. Stokes [1], Jeremy Keane [1], Sarinder Chahal [1], Caroline Samer [1], Miguel Agoncillo [1], Josephine Yu [1], Jennifer Chen [1], Michael Downes 2, Ronald M. Evans 2, Christopher Liddle [1],2, Jenny E. Gunton [1] [1]Centre for Diabetes, Obesity and Endocrinology, The Westmead Institute for Medical Research, University of Sydney, Westmead, NSW, Australia 2 Salk Institute for Biological Studies, La Jolla, CA. USA Background: Vitamin D signals through the vitamin D receptor (VDR) to induce its end-organ effects. Hepatic stellate cells control development of liver fibrosis in response to stressors and vitamin D signaling decreases fibrogenesis. VDR expression in hepatocytes, however, is low in healthy liver, and the role of VDR in hepatocyte proliferation is unclear. Methods: In these studies, hepatocyte-VDR null mice (hVDR) were used to assess the role of VDR and vitamin D signaling in hepatic regeneration. Mice underwent 2/3 partial hepatectomy and liver regeneration and function were studied. Results: hVDR mice had impaired liver regeneration with impaired hepatocyte proliferation. This was associated with significant differential changes in bile salts. Notably, mice lacking hepatocyte VDR had significant increases in expression of conjugated bile acids after partial hepatectomy, consistent with failure to normalize hepatic function by the 14-day time point tested. CDCA (chenodeoxycholate) is an FXR ligand, which was increased after surgery in controls but not in hVDR mice. FXR signaling is important for hepatocyte proliferation. Real-time PCR of hVDR and control livers showed significant changes in expression of cell cycle genes including cyclins D1 and E1 and cyclin-dependent kinase 2. Gene expression profiling of hepatocytes treated with vitamin D or control showed regulation of groups of genes involved in liver proliferation, hepatitis, liver hyperplasia / hyperproliferation and liver necrosis / cell death. Conclusions: Together these studies demonstrate an important functional role for VDR in hepatocytes during liver regeneration. Combined with the known profibrotic effects of impaired VDR signaling in stellate cells, these studies provide a mechanism whereby vitamin D deficiency would both reduce hepatocyte proliferation and permit fibrosis, leading to significant liver compromise.

Presentation: 6/2/2024
8805 Nucleostemin is a novel bifunctional transcriptional coregulator of theandrogenreceptorin prostate cancer cells
Authors: Cheung Edwin , Zhang Cuiting
Published: October 2024

Disclosure: E. Cheung: None. C. Zhang: None.

Androgen receptor (AR) is a master transcriptional regulator in prostate cancer, interacting with and recruiting distinct sets of coregulator proteins to mediate the expression of direct target genes. What are the coregulators of AR and how they function in AR transcriptional activity and prostate cancer biology are still unclear. Using chromatin-immunoprecipitation coupled with mass spectrometry, we identified a set of AR interacting proteins that are present in multiple prostate cancer cell lines. We provide biochemical, cell-based, and genomic evidence to show that Nucleostemin, also known as Guanine nucleotide-binding protein-like 3, is a novel AR coregulator. Specifically, we demonstrate Nucleostemin has dual coregulator activities, functioning as both a coactivator and a corepressor of AR-dependent transcription to regulate the expression of genes that are involved for prostate cancer progression and immune response. From cellular and animal studies, we found that inhibiting Nucleostemin sensitizes prostate cancer cells to Enzalutamide. Clinical analyses of Nucleostemin from prostate cancer patient cohorts revealed that it is expressed significantly higher in cancer than normal in tissues. Finally, we found Nucleostemin expression is an excellent predictor of disease-free survival. In conclusion, our work suggests that Nucleostemin is a novel AR coregulator and is a promising diagnostic and therapeutic target for prostate cancer treatment.

Presentation: 6/2/2024
8516 Role of ER, PR, IRS-1 Transcriptional Complexes in Endocrine Resistant Luminal Breast Cancer
Authors: Schmidt S , Lange C A
Published: October 2024

Disclosure: S. Schmidt: None. C.A. Lange: Consulting Fee; Self; Consultant for Astrin.

In the United States, 1 in 8 women will develop invasive BC within their lifetime. For Luminal breast cancer expressing steroid hormone receptors (SRs) for estrogen and progesterone (ER+/PR+), current standard of care targets ER with antagonists, such as tamoxifen, or blocks estrogen synthesis using aromatase inhibitors. Unfortunately, these endocrine therapies ultimately fail in up to 40% of patients with node positive status. Most endocrine resistant breast cancers retain expression of ER and PR, PGR being a key ER target gene. Two common mechanisms of resistance are ESR1 gene mutations, providing ligand-independent activation of ER, high expression of PR, and activation of oncogenic signaling pathways that circumvent therapies. These pathways include mitogen- and stress-induced signaling pathways (MAPKs, CDKs, PI3K/AKT). Notably, PR target genes encode many of the key components of oncogenic signaling pathways, such as ErbB family members and their ligands. Interestingly, PR can act to limit or divert ER-driven proliferation in hormone-sensitive breast cancer models. However, tumor-promoting actions of PR have been identified by the Lange lab and others. Constitutive ER/PR complexes cooperate at cancer-relevant target genes and both progestins and anti-progestins block estrogen-driven proliferation and survival. The Lange lab demonstrated that MAPK-activated phospho-PRs drive tamoxifen resistance and cancer stem cell expansion. Tumor heterogeneity underscores the need to target both hormone sensitive proliferating cells and non-proliferating cancer stem cells to avoid recurrence of newly resistant BC. A cytoplasmic adaptor molecule in the IGF/insulin signaling pathway, insulin receptor substrate-one (IRS-1), has been identified as a new possible target. We previously reported that IRS-1 is a ligand-independent phospho-PR target gene. Once expressed, IRS-1 forms a complex with ER/PR and is required for breast cancer stemness properties. We hypothesize that the transcriptional regulation of target genes by PR/ER/IRS-1 promotes endocrine resistance and supports stemness phenotypes in ER+ breast cancer. Further studies confirmed a close-proximity nuclear interaction between PR and IRS-1 upon estrogen (but not progestin) stimulation in additional models. This interaction was blocked upon IRS-1 inhibition or insulin treatment. Canonical cytoplasmic insulin/IR/IRS-1 signaling may sequester IRS-1 away from nuclear ER/PR complexes. Accordingly, insulin treatment dramatically reprogrammed the estrogen-driven transcriptome. Characterization of the signal-specific roles of cytoplasmic (insulin) or nuclear (estrogen) IRS-1 may reveal how altered SR signaling complexes contribute to endocrine resistance and may lead to new therapeutic targets for improved outcomes for women with endocrine resistant ER+ breast cancer.

Presentation: 6/3/2024
7587 The Estrogen Receptors and ARE/Nrf2 are Involved in Protecting Human Dermal Fibroblasts from Damage Caused by Mitochondrial Disfunction
Authors: Darawshe A , Trachtenberg A , Sharoni Y
Published: October 2024

Disclosure: A. Darawshe: None. A. Trachtenberg: None. Y. Sharoni: None.

Skin ageing is influenced by several factors including environmental exposure and hormonal changes. Reactive oxygen species (ROS), which mediate many of the effects of these factors, can be formed by extrinsic factors, such as sun exposure, or can result from mitochondrial dysfunction as occurs during ageing. Several studies have shown the protective role of estrogens on skin health. The aim of the current study was to examine the damage to dermal fibroblasts by mitochondrially generated ROS, and to study the mechanism of the protective effects of estradiol. Rotenone, a complex I inhibitor, was used to cause mitochondrial dysfunction in human dermal fibroblasts and its effects on mitochondrial and cytosolic ROS levels, mitochondrial respiration, cell death, apoptosis, matrix metalloproteinase-1 (MMP1) and pro-collagen secretion were determined as markers of skin damage. Rotenone caused substantial reduction of respiration, followed by increased mitochondrial and cytosolic ROS which resulted in apoptotic cell death, increased MMP1 secretion and decreased collagen secretion. Pretreatment with estradiol recovered more than 50% of the respiratory activity, reduced mitochondrial and cytosolic ROS levels and MMP1 secretion and increased cell number and collagen secretion. These effects can be partially explained by a cooperative effect of estradiol and rotenone on Antioxidant Response Element (ARE/Nrf2) transcriptional activity, which leads to upregulation of antioxidant proteins such as NQO1 and Trxr1. To determine if estrogen receptors (ER) are involved in the protective effects, we used the ER inhibitor, Fulvestrant. This inhibitor, which completely inhibited ER Response Element reporter activity, partially prevented the protective effects of estradiol. The protective effects of estradiol were Similarly reduced by ML385 and Ochratoxin A, that are inhibitors of the ARE/Nrf2 transcription system. Incubating the cells with Fulvestrant and Ochratoxin A, which inhibit both pathways, completely blocked the protective effect of estradiol. These results suggest that the estrogen receptors and the ARE/Nrf2 signaling pathways are involved in the protective effects of estradiol against the damage caused by rotenone-induced mitochondria generated oxidative stress.

Presentation: 6/3/2024
8242 A Case of Ayme Gripp Syndrome
Authors: Jaisinghani P , Pappas J
Published: October 2024

Disclosure: P. Jaisinghani: Consulting Fee; Self; Novo Nordisk. J. Pappas: None.

We present a 35 year old female with cataracts, generalized seizures and oligodontia, referred to us for hair loss and elevated TSH. For over 10 years, patient had a diffuse hair loss which progressed to a focal patch of hair loss in the setting of prior use of keppra and seborrheic dermatitis. Patient has regular menses, and no symptoms of hyper/hypothyroidism or family history of thyroid disease. Patient established care with dermatology and scalp biopsy noted early androgenetic alopecia. Physical exam was notable for short stature, height 4'11, 98 lbs, BMI 19.8 kg/m², progeric appearance, facial hyperpigmented spot, hair thinning, palpable thyroid R>L. Laboratory evaluation included TSH 6.150 (0.450 - 4.500 uIU/mL), DHEAS 263 (84.8 - 378.0 ug/dL), total testosterone 15 (10 - 55 ng/dL), FSH 6.6 (Follicular phase 3.5 - 12.5 mIU/mL), LH 7.6 (Follicular phase 2.4 - 12.5 mIU/mL), estradiol 83 (Follicular phase 12.5 - 166.0 pg/mL). Thyroid US demonstrated smaller than average size left lobe measuring 2.93 cm x 0.84 cm x 1.36cm vs right lobe measured 4.34 cm x 0.90 cm x 1.62. Labs noted TSH 5.690 (0.450 - 4.500 uIU/mL), establishing subclinical hypothyroidism and negative thyroid antibodies. Clinical genetics evaluation included trio exome sequencing which revealed a de novo pathogenic variant p.(Ser54Leu) (TCG>TTG): c.161 C>T in exon 1 of the MAF gene (NM_005360.4) which establishes the diagnosis of a Ayme- Gripp syndrome (AGS). AGS is characterized by a triad of cataract, sensorineural hearing loss, and characteristic facial features in combination with neurodevelopmental abnormalities. 21 affected individuals from 19 families have been reported with AGS. Clinical features in our case characteristic of AGS included short stature, brachycephaly, midface hypoplasia, long philtrum and premature hair loss. Hypothyroidism was identified in childhood in 3/15 identified individuals who responded to thyroid hormone replacement therapy. For this patient, we will monitor thyroid function tests every 3-6 months and monitor for symptoms of hypothyroidism. Patient has been referred to cardiology and nephrology, and will continue to follow with genetics and dermatology. Our case further establishes the association of hypothyroidism with AGS. Our case demonstrates the value of expanding the differential of endocrinopathies to rare genetic conditions which are currently identifiable by clinically available genomic tests.

Presentation: 6/1/2024
7165 A Case of Severe Myxedema Coma with Levothyroxine Allergy
Authors: Rothstein Costris A , Yaseen A , Levy S
Published: October 2024

Disclosure: A. Rothstein Costris: None. A. Yaseen: None. S. Levy: None.

Introduction: Myxedema coma is a life threatening condition with a mortality rate of 25-60%. Untreated, it can lead to fatal physiological changes. The treatment is thyroid hormone replacement therapy, alongside supportive care for complications. Clinical Case: A 65-year-old woman, who has been dealing with untreated hypothyroidism since 2012 and reported allergies to various levothyroxine brands, presented with falls, weakness, and exacerbated leg numbness. During the examination, she exhibited periorbital edema, non-pitting edema in the lower extremities, bradycardia, and hypothermia. The blood work revealed an elevation in TSH levels at 301 uIU/L (0.45-5.33 uIU/mL). The patient was administered IV Synthroid (200 mcg), Atropine, and Hydrocortisone due to concerns about myxedema coma. After four hours, the patient experienced angioedema and hypotension, necessitating intubation. This situation was complicated by a pulseless electrical activity (PEA) arrest. Following cardiopulmonary resuscitation and the administration of epinephrine, the patient achieved return of spontaneous circulation. The allergy team assessed the patient for potential anaphylactic shock from IV Synthroid. Subsequently, the patient was initiated onIV Liothyronine and Hydrocortisone, leading to improvement in mental status over a period of a few weeks, and no adverse effects were reported. Thyroid labs improved to TSH of 53 (0.45-5.33 uIU/mL), FT3 of 3.3 (2.5-4.4 pg/mL) and total T3 of 133 (87-178 ng/dl). Upon discharge, the patient was prescribed Liothyronine, with plans to perform a Levothyroxine desensitization test in the outpatient setting. Conclusion: Myxedema coma is an endocrine emergency. Patients usually present with deteriorating mental status, hypothermia, and multiple organ system abnormalities. The diagnosis relies more on the clinical presentation and physical examination than on laboratory results. Thyroid replacement therapy serves as the cornerstone of treatment, with a preference for T4 due to its greater availability and lower likelihood of adverse events compared to Liothyronine, which may increase the risk of mortality at higher doses. Despite associated risks, incorporating low doses of Liothyronine is still considered in treatment plans, driven by concerns about reduced T4 to T3 conversion in myxedema patients. Hypersensitivity reactions to Levothyroxine are uncommon but may manifest as widespread hives, swelling, eczema-like skin rashes, elevated body temperature, and compromised liver function, with only one case reporting an anaphylactic reaction to Levothyroxine. In summary, the potential for experiencing an anaphylactic reaction to Levothyroxine exists, highlighting the need to contemplate alternative treatments, such as Liothyronine. Desensitization methods can be considered once the patient's condition is stable.

Presentation: 6/2/2024