bluuebunny/crossref_metadata_2025 huggingface dataset:
ODP061 When cortisol and catecholamines combine in a severe endocrine condition
Authors: Webster John , Mortensen Michael , Alsayed Mahmoud , Guo Rong , Correa Ricardo
Published: November 2022

Introduction

While the majority of ACTH dependent Cushing's Syndrome cases are the result of over secretion of ACTH from the pituitary gland, ectopic ACTH production can occur. Pheochromocytoma, a tumor originating from the adrenal chromaffin cells and classically associated with catecholamine excess, has also been very rarely shown to secrete ACTH. A 54 year old male with a history of obesity, type 2 diabetes, and resistant HTN presented to ED with a tonic clonic seizure, hypoxia and altered mental status. Labs were notable for thrombocytopenia and hypokalemia. His thrombocytopenia was treated with IVIG and solumedrol. Renin activity and renal doppler were normal. He was discharged on additional blood pressure medication, antipsychotics and Keppra. After 2 weeks he was readmitted with hypertensive urgency, anemia and worsening thrombocytopenia. Bone marrow biopsy did not elucidate the etiology of thrombocytopenia. CT abdomen showed a 5.4 cm, 54 Hounsfield unit, right adrenal mass. MRI brain showed no intracranial pathology. MRI abdomen/pelvis showed bilateral adrenal thickening. The right adrenal mass had characteristics of hematoma as well. Additional labs showed urine metanephrines 1412mcg/24Hr and normetanephrines 978mcg/24Hr, Urine free cortisol was 4494 mcg/24h, random serum cortisol was 207mg/dl, and ACTH was 1091 pg/ml. Dexamethasone suppression test cortisol was 151 ug/dL. Interesting the patient lacked cushingoid phenotypical characteristicsexcept for truncal obesity. The patient was placed on doxazosin for alpha blockadeand etomidate drip to control cortisol. The diagnosis of ACTH secreting pheochromocytoma was reaffirmed after both octreoscan and dotatate PET CT showed the presence of somatostatin receptors in the rightadrenal mass. After 14 days with controlling treatment, the patient underwent successful right adrenalectomy. Pathology was notable for a well circumscribed dark brown mass that had replaced the adrenal medulla that stained patchy positive for ACTH and was positive for synaptophysin and chromogranin. Post surgical labs demonstrated cortisol of 26.3ug/dLand ACTH of 7pg/ml. The patient was discharged 14 days after surgery.

Discussion and conclusion

Pheochromocytomas are rare tumors for which classical features are often absent. Symptoms of palpitations, headache and sweating are only present in 17% of patients. ACTH secreting pheochromocytomas represent 5% of ectopic ACTH syndrome with less than 100 documented cases worldwide. While obesity, hypertension and diabetes are common in our society, this patient's rapid clinical deterioration was the key clue to this rare diagnosis. This case highlights how the decisive actions of a well-coordinated, multidisciplinary team can result in a good outcome for a very challenging and critically ill patient.

Presentation: No date and time listed
ODP062 Clinical Outcomes of Cardiogenic Shock Hospitalization Among Patients with Long-term Systemic Steroid Use
Authors: Kumi Dennis D , Ramirez Marcelo , Soon-shiong Raquel , Patel Birju , Nissan Ninos , Karki Sadichhya , Shrestha Prajwal
Published: November 2022

Background

Cardiogenic shock is a state of immense stress, and our innate human response depends on a robust metabolic, neuroendocrine, and immunologic reserves. Long term use of systemic corticosteroids takes its toll on all aspects of this mechanism. Knowledge of how this basic physiology translates into clinical outcomes is scarce among patients with cardiogenic shock. Our study sought to identify such effects. Method: A retrospective cohort study was designed using data obtained from the 2016 to 2018 combined National Inpatient Sample (NIS) database. Adult patients (age >18) with principal admission diagnosis of cardiogenic shock were identified using the international diseases classification code, tenth revision (ICD-10), and separated into two cohorts based on whether they had a secondary diagnosis of long term systemic steroid use. Primary outcomes assessed were, mortality, length of stay (LOS) and total hospital charge. Secondary outcomes included rate of mechanical ventilation, mechanical circulatory support device use and cardiac arrest. Multivariate linear and logistic regressions were used to adjust for confounders.

Results

There was a total of 477,695 adult hospitalizations for cardiogenic shock, out of which 1. 06% had associated secondary diagnosis of long-term systemic steroid use. The presence of long-term systemic steroid use was associated with 15% increased odds of mortality, (36.63% vs 33.32%, AOR: 1.15, 95% CI: 1. 01 to 1.30, p: 0. 039). However, there was a significant reduction in both length of stay (LOS), (9.85 days vs 11.56 days, adjusted mean difference: -1.59 days, 95%CI: -2.41 days to - 0.77 days, p < 0. 001) and total charge (178,624.8 USD vs 240,859.3 USD, adjusted difference of -55,719.4 USD, 95% CI: - 70,348. 0 to -41,090.8) among patient with long-term systemic steroid use compared to the other cohort. Similarly, there was significantly reduced odds of both the rate of mechanical circulatory support device utilization (15.43% vs 20.32%, AOR: 0.99, 95%CI: 0.62 to 0.77, p: 0. 001) and rate of cardiac arrest (7.22% vs 9.31%, AOR: 0.78, 95%CI: 0.61 to 0.98, p: 0. 035) among patients with long term systemic steroid use. There was no difference in the rate of mechanical ventilation between the two groups.

Conclusion

the presence of long-term systemic steroid use as a comorbidity during hospitalization for cardiogenic shock was associated with an increased mortality but a reduction of LOS, total hospital charge, mechanical circulatory support device utilization and cardiac arrest. One can only hypothesize that the antiparallel relationship between mortality and the other primary and secondary outcomes could be brought about by earlier discharge from hospital due to death, thus truncating further needs for hospitalization and other interventions. This crucial paradox needs further studies to ascertain.

Presentation: No date and time listed
OR25-1 Bilateral and Unilateral Malformations of Male Reproductive Tracts in Mice with Androgen Receptor Gene Deletion in the Mesenchyme
Authors: Jia Shuai , Zhao Fei , Wilbourne Jillian
Published: November 2022

Congenital unilateral defects of male reproductive tracts have been documented in men, suggesting the potential left-right asymmetry in male reproductive tract development. However, classic textbooks of endocrinology have been teaching us that the development of the paired male reproductive tract is governed by the universal action of the androgen receptor (Ar). During sexual differentiation, testis-derived androgens activate the Ar to promote the stabilization of the bilateral Wolffian ducts, the progenitor for the male reproductive tract. The Wolffian ducts then differentiate into the paired male reproductive tract organs, which include the epididymes, vas deferentes, and seminal vesicles. The Ar is expressed both in the epithelium and mesenchyme of the male reproductive tract during fetal development. Landmark tissue recombinant studies and the observation of normal morphology in the epithelium-specific Ar knockout mouse all support the notion that Ar action in the mesenchyme dictates the morphogenesis of the male reproductive tract. However, no genetic study has been performed to investigate the consequence of the loss of mesenchymal Ar on male reproductive tract development. To test the functional significance of the mesenchymal Ar, we designed a mesenchyme-specific Ar knockout mouse model (ARcKO) to ablate Ar specifically in the mesenchyme prior to the initiation of sexual differentiation. We performed immunohistochemistry of AR to confirm that Ar expression was absent in the mesenchyme while epithelial Ar remained intact in this ARcKO model. Based on the widely accepted notion that mesenchymal Ar regulates epithelial morphogenesis of the male reproductive tract, we expected to see abnormal patterning of the male reproductive tract. Indeed, the epididymis lost its characteristic coiling and became cystic. However, 40% of the 23 collected ARcKO males displayed these abnormalities on both horns; 43% and 17% displayed these abnormal phenotype solely on the left and right horn, respectively. This surprising observation might result from asynchronous activities of Cre on the left and right horns. However, when the Cre was crossed with a reporter (tdTomato), we observed comparable reporter expression on both horns at the onset of sexual differentiation of reproductive tracts. These observations suggest that the developmental programs in the morphogenesis of the left and right male reproductive tracts are asymmetrical and there could be a compensation mechanism for the loss of mesenchymal AR. We are currently in the process of comparing the left and the right horns in Ar knockout male mice to determine any biased signaling. Taken together, our study provides a unique model for not only studying congenital defects of male reproductive tracts but also for investigating the potential asymmetry in the masculinization program.

Presentation: Monday, June 13, 2022 11:00 a.m. - 11:15 a.m.
PMON201 The Impact of Menopause on Poor Outcomes in Hospitalized Patients with COVID-19 Infection
Authors: Toure Tonslyn , Ravindra Lakshmi , Monteiro Filipe , Gopalakrishnan Geetha
Published: November 2022

Background

Studies evaluating risk factors for poor outcomes with COVID-19 infection report gender variation. Impact of estrogen on immune regulation may be one factor contributing to better outcomes in women. This study evaluates the impact of menopausal status and other risk factors on health outcomes in women hospitalized with COVID-19 infection.

Methods

Data from 1863 non-pregnant women admitted to the Lifespan Hospital System in Rhode Island with COVID-19 infection during the first wave (March 1 - June 30, 2020) and second wave (July 1, 2020 - February 28, 2021) were analyzed. Subjects were categorized as pre-menopausal (age less than 45 years), perimenopausal (age 45 to 55 years), and post-menopausal (age greater than 55 years). Multivariate models to look for risk factors associated with severe illness (i.e. intensive care unit admissions, mechanical ventilation and death) controlled for menopause status, race/ethnicity, insurance type, wave of COVID-19, body mass index, diabetes (DM), hyperglycemia, hypertension (HTN), pulmonary disease, cardiovascular disease (CVD) and chronic kidney disease (CKD).

Results

In our cohort, pre-, peri- and pre, peri and post- menopausal status represented 12.8, 10.4 and 76.8% of the study population, respectively. The mean (± SD) age was 67.57 ±18.0 with 61.4% White/Caucasian, 20% Hispanic/Latino and 1.7% Black. 98.4% had commercial or government-sponsored health insurance. Hyperglycemia (Blood glucose > 180mg/dL on two occurrences) was noted in 26.8% of hospitalized women. Compared to pre- and peri- menopausal status, post- menopausal status was associated with a higher prevalence of Hyperglycemia (OR 1.59 [1.23-2.07]), DM 1.90 (1.50-2.42), HTN 5.06 (4.02-6.37), Pulmonary disease 1.35 (1.07-1.70) and CKD 3.7 (2.58-5.42). Post-menopausal status was also associated with higher mortality (OR 8.6 [2.7-27.6]), severe illness (OR 5.7 [1.3-23.9]), readmission (OR 1.5 [1.04-2.2]) and longer length of hospital stay (OR 1.6 [1.2-2.2]). Adjusted model identified, hyperglycemia (OR 14.8 [4.5-48.8]) and first wave (OR 2.7 [1.1-6.4]) to be independent risk factors for severe illness in post-menopausal women admitted to the hospital with COVID -19 infection.

Conclusion

While patients with comorbidities are known to be at greater risk for complications related to COVID-19 infection, menopausal status has rarely been considered a risk factor. Recognizing underlying risk factors in females, particularly menopausal females with COVID-19, can help clinicians attempt to identify patients at a higher risk of COVID-19 complications and drive early, more aggressive interventions to mitigate risk in this population.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON224 Newly Observed Interference of Oral Biotin with the Free Testosterone Assay? In a Woman with Postmenopausal Virilization.
Authors: Caputi Rafael , Ayala Alejandro
Published: November 2022

Introduction

Approximately 15-20% of US adults take supraphysiologic doses of biotin-containing supplements for common hair and skin problems. When consumed in high doses, competitive and non-competitive immunoassays which use the streptavidin-biotin binding complex can skew results to be either falsely high or falsely low.

Case Presentation

A 74 y/o Caucasian female who initially presented to her primary care physician concerned for alopecia with frontal balding, acne and moderate hirsutism. Her total testosterone was elevated with 146 ng/dL (7.0–40 ng/dL) with a perplexing normal free testosterone of 4.2 pg/mL (<4.2 pg/mL) in the context of a normal SHBG of 48 nmol/L (14–73 nmol/L). Androstenedione was elevated with 161 ng/dL (17–99 ng/dL) with a normal CMP, prolactin, DHEA-s and 17-hydroxyprogesterone. Transvaginal US was normal. The size of the ovaries was consistent with menopause. Abdominal and pelvic MRI showed normal adrenal glands and ovaries. She was taking between 1 to 3mg of biotin per day. Biotin was discontinued. Repeat testing revealed an elevated free testosterone of 12.7 pg/mL (0.2 - 3.7 pg/mL) along with an elevated total testosterone of 120 ng/dL (2 - 45 ng/dL) and a normal SHBG of 39 nmol/L (14–73 nmol/L) which was now consistent with her clinical picture of postmenopausal hyperandrogenism. Bilateral laparoscopic salpingo-oophorectomy was recommended to exclude an androgen-secreting ovarian tumor or hyperthecosis. Testing was then repeated while taking biotin which again manifested a normal free testosterone of 2.8 pg/mL (<4.2 pg/mL) with an elevated total testosterone of 121.4 ng/dL (7.0–40 ng/dL) and a normal SHBG of 45 nmol/L (14–73 nmol/L).

Discussion

Biotin may interfere with the total testosterone levels, but little is known of its effects on free testosterone. We report the first case of biotin interference on the free testosterone assay, resulting in falsely low values demonstrated in two different occasions, that improved following withdrawal of this widely used vitamin supplement. Laboratories use a direct analog enzyme immunoassay to calculate the free testosterone however its calibration and specificity has been debatable, and it is still unclear how biotin can affect it. It is critical that laboratorians and clinicians are aware of this biotin interference so that misdiagnosis and inappropriate treatment can be prevented. References: 1. Macarena Alpañés, José M. González-Casbas, Juan Sánchez, Héctor Pián, Héctor F. Escobar-Morreale, Management of Postmenopausal Virilization, The Journal of Clinical Endocrinology & Metabolism, Volume 97, Issue 8, 1 August 2012, Pages 2584–2588.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON225 Androgen excess in pre-menopausal women due to Granulosa Cell tumor: A case report.
Authors: Kaval Roopitha , Moll Teresa , Shenoy Veena , Alhussain Aljunaid , Ridway Mildred , Crews James , Cardozo Licy Yanes
Published: November 2022

A 23-year-old nulliparous female without any significant past medical history presented for irregular menstrual cycles, amenorrhea, hirsutism and 25-pound weight loss in one year. Excessive hair growth was found on the face and chest. Prior to endocrinology visit she was started on oral contraceptives without any improvements. Of importance, she had a negative progesterone challenge test (no withdrawal bleeding). On examination in the Endocrinology clinic, she was a class II morbidly obese female, with coarse terminal hair on her chin and chest and no cushingoid features. Intra-vaginal ultrasound revealed large bilateral multi cystic masses in the ovaries. Her laboratory work up was significant for elevated DHEA-S at 826.6 ng/dl (134-407), Total testosterone 144 ng/dl(0-60) and free testosterone 4.75 ng/dl (0.06-1.08). Her 17-OH pregnenolone was also elevated at 539 ng/dl (31- 455), Inhibin B was elevated at 374 pg/ml (<139 premenopausal). 17 - OH progesterone was normal. Estradiol, inhibin A and FSH/LH were normal. ACTH, AM cortisol, TSH and prolactin were normal. Beta HCG and Cancer markers CA 19-9, CA 125, and CEA were negative. CT scan of the abdomen showed a right ovarian mass measuring 10.7×9.6×9.2 cm. Adrenal glands were normal. Patient subsequently underwent a laparoscopic diagnostic right salpingo-oophorectomy. Pathology was reported as an adult granulosa cell tumor (9.5 cm) of the ovary with an intact capsule. After surgery Inhibin B level decreased from 374 to 10 (normal <139), Total testosterone decreased from 144 to 36 ng/dl (0-60), Free testosterone decreased from 4.75 to 1.22 ng/dl (0.06-1.08) and DHEA-S from 826.60 to 687.40 ng/dl (134-407) however remained elevated. Moreover, after surgery irregular menses resolved and she noted some improvement in hirsutism.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON230 Pancreatic Exocrine Cells are Altered during the Progression of Ovarian Cancer-Driven Cachexia in Female Mice
Authors: Elesh Ibrahim , Kim So-Youn , Lele Subodh , Luan Yi , Yu Seok-Yeong
Published: November 2022

Cancer-associated cachexia (CAC) is a progressive debilitating syndrome that frequently complicates metabolic function in body vital organs and subsequently causes a high mortality rate in cancer patients. However, the underlying mechanisms and the functional consequences of organs dysfunction during CAC are not fully elucidated. Recently, we have developed a preclinical model characterized by oocyte-specific knockin for constitutively active Pi3k (Pik3ca*) which induces ovarian granulosa cell tumor (GCT) and mimics the stagewise progression of CAC in humans. Here, we aim to characterize the pathophysiological effect of cachexia on pancreatic dysfunction and investigate the potential role of the pancreas in the development and complication of cachexia in transgenic female mice with ovarian cancer. Notably, we found that cachectic mice showed a drastic loss in the pancreatic volume (PV) before death. Histological and morphometric assays revealed that exocrine pancreatic acinar atrophy (PAA) is drastic in the pancreas of cachectic PIK3CA* mice. The pancreas appeared to have an increased number of pancreatic stellate-shaped cells between acini. No significant changes were observed in the morphology of endocrine islets. Immunohistochemical assay confirmed a high level of TNF-a in atrophied pancreatic tissue and decreased intensity of pancreatic amylase in the cytoplasm of atrophied pancreatic acinar cells. Additionally, moderate fibrosis was also observed in the wall of pancreatic ducts and blood vessels, but not between the pancreatic acini. Masson trichrome and histochemical staining confirmed expression of extracellular matrix proteins including collagen I, II, IV, and a -smooth muscle actin in ducts and vessel walls. Overall, our preliminary findings in this novel CAC mouse model strongly suggest potential signs of exhausted exocrine pancreatic cells during the progression of ovarian cancer-driven cachexia. This mouse line could be an ideal preclinical model to aid in clarifying molecular mediators of pancreatic pathophysiology and metabolic dysfunction during CAC.

Keywords: Ovarian cancer-driven cachexia, pancreas, atrophy, amylase.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON238 Bone mineral density in middle-aged women with reproductive disorders
Authors: van Zwol - Janssens Charissa , Hage Aglaia , van der Ham K , Velthuis B K , Budde R P J , Koster M P H , Louwers Y V , Franx A , Fauser B C J M , Boersma E , Laven J S E
Published: November 2022

Objective

Reproductive disorders, such as premature ovarian insufficiency (POI) and polycystic ovary syndrome (PCOS), are associated with hormonal and metabolic changes, which might lead to changes in bone mineral density (BMD). On the one hand women with POI enter menopause too early and have less estrogen exposure rendering them more prone to develop low BMD. On the other hand women with PCOS, who often have a delayed menopause and therefore experience longer estrogen exposure, may have an increased BMD. A third reproductive disorder, preeclampsia (PE) does not seem to affect BMD or menopausal age. The effect of these reproductive disorders, with different menopausal ages, on BMD could be most evident in middle-aged women. We hypothesized that the duration of estrogen exposure is a key factor in BMD.

Materials and methods

In this cross-sectional study we compared the BMD of 258 women with a history of PE as a reference group (age 46.0; 40% postmenopausal) to 90 women with POI (age 48.1; 100% postmenopausal) and 84 women with PCOS (age 48.3; 26% postmenopausal). BMD was measured using non-contrast coronary CT scans in three consecutive thoracic vertebrae (in the T7-T10 range). Secondary outcomes included T-scores, prevalence of osteoporosis and osteopenia and the comparison of measured BMD to predicted BMD, calculated by a formula based on age. We performed linear regression analysis to assess differences in BMD between groups, adjusted for age and BMI.

Results

Compared to women with a history of PE, women with POI had lower BMD (p<0.001) and women with PCOS had higher BMD (p=0.003). Prevalence of osteoporosis was higher in women with POI (12.2%) compared to PE (0.8%; P=0.002) and none of the women with PCOS fulfilled the criteria for osteoporosis. Prevalence of osteopenia was significantly higher in the POI group and lower in the PCOS group compared to the PE group (57.8%, 17.9% and 26.4% respectively; p<0.01). As expected, measured BMD of women with POI was significantly lower than the predicted BMD. On the contrary, the measured BMD of women with PCOS was significantly higher than the predicted BMD. Interestingly, the measured BMD of women with a history of PE was also higher than the predicted BMD.

Conclusion

Our results imply that prolonged estrogen exposure as in PCOS is associated with a higher BMD. A lower estrogen exposure associated with an early menopause in POI is associated with a lower BMD and osteoporosis. Patients with a history of PE could be seen as an intermediate group in terms of BMD and estrogen exposure, however, the measured BMD of women with a history of PE was higher than the predicted BMD. This study emphasizes the importance of adequate counseling on long term health issues when diagnosing a reproductive disorder.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON246 Prevalence of Hypertension and Metabolic Syndrome in Women with PCOS from France and Brazil
Authors: Spritzer Poli Mara , Marchesan Lucas B , Perol Sandrine , Allgayer Roberta M , Plu-Bureau Geneviève
Published: November 2022

Polycystic ovary syndrome (PCOS) is a common endocrine disorder, affecting women of reproductive age and has been associated with metabolic and reproductive abnormalities. Evidence from different geographic regions indicates that the metabolic features of PCOS may be influenced by ethnicity, lifestyle and sociocultural backgrounds. Also, metabolic disturbances seem to be found in higher frequency in referral than in unselected populations and from a clinical practice point of view, data from referral samples may be relevant for adjusting effective primary and secondary prevention strategies. Therefore, the aim of this study was to determine the prevalence of systemic arterial hypertension and metabolic syndrome (MBS) in women with PCOS from France and Brazil. We retrospectively assessed 314 French (FR) women and 236 Brazilian (BR) women with PCOS, according to the Rotterdam criteria, aging between 18 and 42 years and not using combined oral contraceptives or metformin. These women had been referred to the respective University Hospitals between 2014 to 2021 in France and 2014 to 2020 in Brazil. BR women had participated in previous studies conducted at our research center. Hypertension was defined according to the ACC/AHA 2017 and MBS according to the Joint Scientific Statement, 2009. Mean age was 26.0 (SD 5.9) years FR and 25.9 (SD 5.8) BR, p=0.87, and mean body mass index (BMI) was 27.1 (SD 6.8) FR and 31.3 (SD 7.5) BR P<0.001. The frequency of classical phenotype (A/B) was 83.7% in FR (n=262) and 77.1% in BR (n=182), p =0.071. Subsequent analyses were done including only women presenting the classical phenotype from each center. FR women with PCOS presented lower prevalence of overweight/obesity (53.7% FR vs. 84.6% BR, p<0.001), hypertension (32.6% FR vs. 71.6% BR, p<0.001) and MBS (12.5% FR vs. 38.5% BR, p< 0.001) than BR women with PCOS. Using logistic regression models for hypertension and MBS, it was found that both BMI and country contributed significantly for these prevalence results. In conclusion, women with PCOS living in France presented a less severe metabolic profile than those living in south Brazil. Differences in dietary pattern, lifestyle, socio-cultural features and ethnicity may have contributed to these results. Further studies are warranted to deeper characterize the influence of dietary composition and lifestyle on prevalence ratios of metabolic features in women living in France.

Sources of Research Support: INCT-CNPq /FAPERGS

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
ODP066 Impact of Comorbid Adrenal Insufficiency on Outcomes of Infective Endocarditis Hospitalizations
Authors: Akaho Elikplim , Kumi Dennis , Ramirez Marcelo , Karki Sadicchya , Shrestha Prajwal , Nissan Ninos , Patel Birju , Soon-shiong Raquel
Published: November 2022

Background

Adrenocortical integrity is vital to mounting an appropriate response to a potentially deleterious endovascular infection such as infectious endocarditis. Likewise, adrenocortical hormones affect how cytokine storms lead to secondhand injuries that complicate sepsis. Data on the clinical outcomes of infective endocarditis in patients with adrenal insufficiency is limited and thus our attempt to study this topic was perhaps a worthwhile venture. Method: A retrospective cohort study was designed using data obtained from the 2016 to 2018 combined National Inpatient Sample (NIS) database. Adult patients (age >18) admitted with a principal diagnosis of acute and subacute infective endocarditis were identified using the international diseases classification code, tenth revision (ICD-10). They were then stratified into two cohorts based on the presence of adrenal insufficiency. Primary outcomes assessed were, mortality, length of stay (LOS) and total hospital charge. Secondary outcomes included septic shock, embolic stroke, cardiogenic shock, and septic arterial embolization. Multivariate linear and logistic regressions were used to adjust for confounders.

Results

There was a total of 36,669.97 adult hospitalizations for infective endocarditis, among which 0.67% had a secondary diagnosis of adrenal insufficiency. The presence of adrenal insufficiency led to an increased length of stay (19.66 days vs 13. 05 days, adjusted mean difference of 7.17 days, 95%CI: 1.41 to 12.92 days, p: 0. 015) and an increased total charge (342,312.3 USD vs 147,887.6 USD, adjusted mean difference: 189,447.6 USD, 95%CI: 32,509.7USD to 346,385.5 USD, p: 0. 018) compared with patients without adrenal insufficiency. There was no significant difference in the odds of mortality between the two groups (4. 00% vs 4. 01%, AOR: 0.87, 95% CI: 0.99 to 7.57, p: 0.898). Similarly, other secondary clinical outcomes including septic shock, cardiogenic shock, septic arterial embolization, and embolic strokes were not different between the two cohort groups.

Conclusion

The presence of adrenal insufficiency among patients admitted with infective endocarditis led to an increased length of stay and a commensurate increased total charge. However, it did not significantly impact the outcomes in terms of mortality, rate of septic shock, cardiogenic shock, septic arterial embolization, or embolic strokes.

Presentation: No date and time listed
PMON267 Comparison Between Single Time Point Testing and 24-Hour Average Concentration of Total Testosterone in Hypogonadal Men Treated with an Oral Testosterone Undecanoate Softgel (JATENZO®)
Authors: Swerdloff Ronald , Wang Christina , Gittelman Marc , Newmark Jay , Woun Seo B , Rohowsky Nestor , Dudley Robert
Published: November 2022

In 2019, a novel, first-in-class testosterone (T) replacement therapy (TRT), oral testosterone undecanoate (TU) was approved by the U.S. FDA for the treatment of male hypogonadism. During clinical trials, dose adjustments were based on 24-hr average T concentration (Cavg) because single T measurements were considered less accurate. Subsequent, concordance analyses for this oral TU product have shown that a single total T value 6-hrs after the morning oral TU dose best corresponds to Cavg. Nonetheless, a conversion factor for T values obtained at other post-dose time points would be useful for healthcare providers (HCPs) and ease potential scheduling challenges for patients. Consequently, the relationship between other T sampling time points and Cavg was examined to determine if a reliable conversion factor could be derived to help HCPs monitor a patient's serum T concentration at times other than 6-hrs after oral TU administration. Hypogonadal men, age 18–65 y/o, were recruited into a randomized, open-label, multicenter, dose-titration trial. Overall, 166 men were randomized into the oral TU arm. Dose titration was based on Cavg calculated from serial pharmacokinetic (PK) samples. There were three pre-designated PK visits, to individualize the appropriate TU dose and to achieve a eugonadal T Cavg by the final study visit. Ratios between different timepoints and Cavg were determined for PK samples following morning drug administration. Overall, 87.3% (95% CI: 81.3%, 92.0%) of hypogonadal men had a final Cavg in the eugonadal range, with a mean serum total T = 488.7 ± 154.5 ng/dL (16.95 ± 5.37 nmol/L). Pooled values from all PK days demonstrated a linear relationship between T concentrations at 4, 6, and 9-hrs and Cavg (p < 0.0001). Visit and time of sampling interaction were not statistically significant (p >> 0.50), indicating consistency in results among PK visits and among sampling timepoints. A factor was derived to enable conversion of T values assessed at a time other than 6-hrs (post oral TU dose) into a close approximation of Cavg: 1/[1.870–0.14×(hours after AM dose)]. Therefore, a sample drawn at 4-hrs after the morning oral TU dose would be multiplied by 0.75; while one drawn at 8-hrs would be multiplied by 1.33. Hence, T Cavg can be approximated after morning JATENZO administration if a blood sample cannot be collected precisely 6-hrs thereafter.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
RF34 | PMON202 The Human Metabolic Profile of Estetrol
Authors: Gérard Céline , Apter Dan , Chatel Guillaume , Nollevaux Fabrice , Jost Maud , Gaspard Ulysse , Foidart Jean-Michel
Published: November 2022

Objectives

Estetrol (E4), a native estrogen produced by the fetal human liver, has four hydroxyl-groups, making it structurally distinct from other estrogens. We determined the human metabolic profile of E4, which is recently marketed combined with drospirenone as a novel contraceptive and is currently in development for the relief of menopausal symptoms.

Method: We studied the in vitro metabolism of E4 using human hepatocytes, human liver microsomes and recombinant CYP enzymes in reaction phenotyping studies to evaluate the role of human cytochrome P450 enzymes, UDP-glucuronosyltransferases (UGT) and sulfotransferases (SULT). Furthermore, we evaluated the in vivo metabolism of E4 in an open-label, non-randomised phase 1 human absorption, distribution, metabolism, and excretion study, in which six healthy postmenopausal participants received a single oral solution containing 15 mg [14C]-E4. We analysed blood (up to 240h), urine and faecal samples (up to 312h). Metabolite profiling and identification was performed using liquid chromatography with tandem mass spectrometry.

Results

The in vitro studies showed that E4 undergoes extensive phase 2 metabolism to form glucuronide and sulphate conjugates. UGT2B7 is the key enzyme catalysing the direct glucuronidation of E4 and SULT1E1 is the dominant sulfotransferase for sulphate conjugation in vitro. Unlike for other estrogens, oxidative phase 1 metabolism by P450 enzymes does not play a major role in the metabolism of E4. Following oral administration of E4 to humans, the major metabolites observed in human plasma were E4-16-glucuronide, E4-3-glucuronide and E4-glucuronide-sulfate (respectively 62%, 17% and 9% of radioactivity at the time of maximum plasma concentration [Tmax]). At Tmax, unchanged E4 accounts for only 7.5% of plasma radioactivity. Main metabolites quantified in urine during the first 6 hours after administration were E4-16-glucuronide and E4-3-glucuronide (respectively 77.4% and 16.5% of urine radioactivity). Approximately 69% of the total administered radioactivity was recovered in urine and 22% in faeces. Mass spectrometry showed that E4 was not present in urine but was detected in faeces and that E4 is not converted back into one of the other natural estrogens (estriol, estradiol, estrone).

Conclusions

Altogether, these data suggest that E4 has a unique metabolic profile. E4 is a terminal end-product of estrogen metabolism, which is not converted back, in vivo, into active metabolites like estriol, estradiol or estrone. Unlike for other estrogens, CYP450 enzymes do not play a major role in the metabolic pathway of E4.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m., Monday, June 13, 2022 12:48 p.m. - 12:53 p.m.
RF34 | PMON203 Copy Number Variant Analysis Reveals Novel Candidate Genes Associated with Primary Ovarian Insufficiency
Authors: Franca Monica , Moraes Daniela , Franca Monica , Krepischi Ana , Faria-Jr Jose , Costa Elaine , Hayashida Sylvia , Maciel Gustavo , Costa Silvia , Kulikowski Leslie , Jorge Alexsander , Mendonca Berenice , Domenice Sorahia
Published: November 2022

Primary ovarian insufficiency (POI) is characterized by a heterogeneous genetic background. Moreover, the etiology of most POI cases remains unclear. Herein, our goal was to investigate the presence of rare deletions or duplications in patients presenting with POI, which could lead to identification of novel chromosomal regions and/or genes potentially associated with disease risk.

Patients and Methods

Sixty-six patients presenting idiopathic POI were evaluated; 44 had primary amenorrhea and 22 secondary amenorrhea. Chromosomal microarray analysis was performed to detect copy number variants (CNVs) using array-CGH or SNP-array. Identified CNVs were validated by population databases of either health individuals (Database of Genomic Variants - DGV) or patients carrying chromosomal structural alterations (DECIPHER). A set of rare CNVs was selected and annotated in regard to gene content and clinical associations using bioinformatic tools (RefSeq, OMIM, PubMed, Ovarian Kaleidoscope Database, and Mouse Genome Informatics).

Results

Fifteen rare CNVs were found in 13 patients, ten of them with primary amenorrhea. The identified CNVs comprised four deleted and 11 duplicated regions, in which the break points occurred intragenic in 8 of them. Most of the CNVs were mapped in the autosomes. Seven CNVs identified in 5 patients were classified as pathogenic or probably pathogenic; three were novel variants containing candidate genes associated with ovarian development [19q13.43 (NLRP5) and Xq13.2 (XIST)] or function [(2q37.3 (PER2)]. The other pathogenic CNVs were previously implicated in POI etiology; two of them [Xq27.2q28 del (14.5 Mb) and Xq22.1q27.1 dup (41.4 Mb)] were encompassed in a complex chromosome rearrangement, associated with 20p13 dup (2.7 Mb), and the last region was 15q25.2 deletion. Four VUS containing genes associated with ovarian development [2p23.1 (XDH and EHD3), 9p24.1 (KDM4C), 22q12.1 (ZNRF3)] or function [1p31.1 (PTGFR)] were identified.

Conclusion

CNVs analysis indicated a genetic etiology in 10% of these POI patients. New candidate genes associated with ovarian development and function were revealed.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m., Monday, June 13, 2022 12:54 p.m. - 12:59 p.m.
ODP435 11-Ketotestosterone is One of the Major Androgens in Pigs
Authors: Yazawa Takashi , Sato Takahiro , Ida Takanori
Published: November 2022

11-ketotestosterone (11-KT) is one of the active androgens, which belongs to the novel 11-oxygenated class of androgens. Although 11-KT has been regarded as a teleost-specific androgen, it was recently demonstrated that 11-KT is a major human androgen. However, the details of its functions and dynamics remain unknown. Deficiency of proper animal model is one of the reasons for this. Although rodent models have contributed to reveal synthesis and functions of steroid hormones, only small amounts of 11-KT are synthesized in mice and rats (Yazawa et al. Endocrinology, 2008). In mammal, 11-KT is abundant in limited animal species, such as non-human primates, pig and guinea pig. This suggest that these animal species are valuable models for determining the functions of 11-KT in mammals. In the present study, we evaluated the production and potential functions of 11-KT in pig. In previous studies, we studied human and murine steroidogenesis using steroidogenic cells-derived from mesenchymal stem cells (MSCs, Yazawa et al. Endocrinology 2006, 2009; Mol Endo, 2010; Endocrine J 2016). Therefore, we first induced steroidogenic cells from porcine subcutaneous preadipocytes (PSPA cells), which originate from MSCs. With the aid of cAMP, adenovirus-mediated introduction of SF-1/Ad4BP induced the differentiation of PSPA cells into 11-KT-producing cells. Using PSPA cell-derived steroidogenic cells and our previously established method (Yazawa et al. JSBMB, 2020), we found that porcine AKR1C1 can convert androstenedione into testosterone (T) and is expressed in adrenal, testis and ovary. In addition, we showed that HSD11B2, one of the essential genes to produce 11-KT from T was expressed in testicular Leydig cells and the adrenal cortex. 11-KT was present in the plasma of both immature male and female pigs, with slightly higher levels in males than in females. T levels were much higher in male pigs. It is noteworthy that 11-KT levels were >10-fold higher than T levels in female pigs. However, castration altered the plasma profiles of 11-KT and T in male pigs to the female-like profiles. 11-KT induced endothelial nitric oxide synthase (eNOS) in porcine vascular endothelial cells. These results indicate that 11-KT is produced in porcine adrenal glands and testes. It can potentially be involved in the regulation of cardiovascular functions through eNOS expression. In addition, these findings indicate that pigs (especially female) represent an ideal model for analyzing the functions of 11-KT in mammal.

Presentation: No date and time listed
ODP441 NErbB-2 and Androgen Receptor Regulate a Gene Signature Involved in Immune Response and Favorable Outcome in TNBC
Authors: Elizalde Patricia , Roldan Deamicis Agustina , Oakley Robert , Madera Santiago , Cordo Russo Rosalía , Schillaci Roxana , Fresno Cristóbal , Cidlowski John , Proietti Cecilia
Published: November 2022

Triple negative breast cancer (TNBC) has poor prognosis and neither established biomarkers nor therapeutic targets. On the one hand, the androgen receptor (AR), a steroid hormone receptor (SR) which is expressed in 10-53% of TNBC and proved to be critical for BC proliferation, has been proposed as a new target in TNBC. On the other, we and others have shown that membrane ErbB-2 migrates to the nucleus (nuclear ErbB-2, NErbB-2) where it binds DNA at HER-2 associated sequences (HAS) to regulate BC proliferation and migration. Since we have previously shown a functional interplay between growth factors and SR signaling pathways in BC, we propose the existence of an interaction between AR and ErbB-2 which is involved in NErbB-2+/AR+ BC growth. The experimental model used was the human TNBC cell line MDA-MB-453 which displays high expression levels of AR and NErbB-2. We have previously shown that dihydrotestosterone (DHT)-activated AR induces Src-mediated ErbB-2 rapid activation and its migration to the nucleus where it binds to HAS sites in the DNA. By coimmunoprecipitation and WB studies, we have now shown that this crosstalk involves ErbB-2 and AR physical association which is inhibited by the AR antagonist enzalutamide. By microarray and bioinformatics analysis we have also previously identified a set of differentially expressed genes (DEGs) in the presence of DHT and NErbB-2 eviction to define an independent predictor of better clinical outcome in TNBC. By using the Tumor Immune Estimation Resource (TIMER) we found a significant association of the expression of the gene signature and the abundance of tumor immune infiltrates, in accordance with the involvement of the gene signature in the immune response and interferon pathways. We have validated the regulation of the gene signature in MDA-MB-453 cells and found that in the presence of DHT, eviction of NErbB-2 significantly induces the mRNA expression of the genes CXCL10, HLA-A, NMI, STAT1 and TAP1, members of said gene signature. We have also identified AREs and HAS sites in STAT1, NMI and TAP1 and studied AR and ErbB-2 recruitment to the chromatin by ChIP assays. We found that DHT induces ErbB-2 binding to its HAS sites in STAT1 in regulatory regions. Our findings evidence that DHT-activated AR induces ErbB-2 rapid activation and its migration to the nucleus where it binds to HAS sites in regulatory regions of a set of genes involved in the immune response in TNBC.

Presentation: No date and time listed
ODP443 Targeting NAD+ availability modulates 11β- Hydroxysteroid dehydrogenase 1 activity in mouse and human cell lines and primary cells
Authors: Kabli Ali , Elhassan Yasir , Heising Silke , Heaselgrave Samuel , Hardy Rowan , Hodson David , Morgan Stuart , Lavery Gareth
Published: November 2022

Four nicotinamide adenine dinucleotide coenzymes – NAD+, NADH, NADP+, and NADPH – are the central catalysts of metabolism. The enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) is NADPH-dependent reductase that catalyses the reduction of the inactive glucocorticoid cortisone) to active cortisol (11-dehydrocorticosterone to corticosterone in rodents). 11β-HSD1 is in the endoplasmic reticulum (ER) lumen, and the level of NADPH in the ER regulates enzyme activity. Little is known about the biochemical mechanisms controlling cytosolic/ER NAD(P)(H) crosstalk. Here we examined how perturbed cellular NAD+ availability modulates 11β-HSD1 enzyme activity in the ER across a range of mouse and human cell types. 11β-HSD1 activity was measured in a series of transformed and primary mouse and human liver (HepG2), muscle, and Human Dermal Fibroblast cells (HDFn) depleted for NAD+ using FK866 to inhibit nicotinamide phosphoribosyl transferase (NAMPT), the rate-limiting enzyme in NAD+ biosynthesis. To replete NAD+ we supplemented cells with 0.5mM of NAD+ precursors nicotinamide riboside (NR) and nicotinamide mononucleotide (NMN). FK866 (100nM, 48h) was effective at depleting NAD+ by approximately 90% in C2C12 myotubes and 50% in HepG2 liver cells. In each cell line, 11β-HSD1 reductase activity was decreased by FK866 compared to untreated controls (80% in C2C12 P<0. 0001, 55% in HepG2, P=0. 03 and 65% in HDFn P=0. 002). Restoring NAD+ level with NR (0.5mM, 6h) in C2C12 and (0.5mM, 24h) in HepG2 and HDFn rescued 11β-HSD1 enzyme activity compared with no precursor treatment (60% in C2C12 P=0. 0007, 52% in HepG2 P=0. 03 and 68% in HDFn P=0. 0006). Human dermal fibroblasts treated with TNFa (10nM, 24h) boosted 11β-HSD1 expression 4-fold, however this was not able to overcome the FK866 induced drop-in reductase activity. These data show that depletion of NAD+ and impaired 11β-HSD1 reductase activity in the ER is a generalised phenomenon across cell types. This suggests a crosstalk between cytosolic and ER NAD(P)(H) pools that can respond rapidly to changes in NAD+ precursor availability. This could provide new insight into the regulation of cellular glucocorticoid metabolism and ER NAD(P)(H) homeostasis.

Presentation: No date and time listed
OR07-1 Estrogen-regulated Enhancer RNAs Contain a Functional Motif that Controls Enhancer Assembly and Function in Breast Cancer Cells
Authors: Hou Tim , Kraus W Lee
Published: November 2022

Approximately 70% of breast cancers express estrogen receptor alpha (ERα), a hormone-regulated transcription factor. Upon binding to endogenous and exogenous ligands, ERα dimerizes and binds to regulatory regions across the genome known as enhancers. Enhancers act as nucleation sites for the recruitment of coregulatory proteins and ultimately regulate target gene transcription. We have previously used global run-on sequencing (GRO-seq) to detect transcription at active enhancers that generates enhancer RNAs (eRNAs). eRNAs are poorly characterized noncoding RNAs and are thought to regulate enhancer function and gene expression. To better understand the functions of these eRNAs, we annotated the estrogen-regulated eRNA transcriptome in ERα-positive breast cancer cells using precision nuclear run-on of capped RNA (PRO-cap) and polyA-depleted RNA-seq. We then cloned selected eRNAs, fused them to single guide RNAs, and targeted them to their ERα enhancers of origin using CRISPR/dCas9. eRNAs promoted expression of cognate, but not heterologous, target genes by increasing ERα recruitment and simulating p300-catalyzed H3K27 acetylation at the enhancer. We identified a ∼40 nucleotide functional eRNA regulatory motif (FERM) present in many estrogen-regulated eRNAs that is necessary and sufficient to promote gene expression by increasing ERα recruitment and stimulating p300-catalyzed H3K27 acetylation. The FERM interacted with BCAS2, an RNA-binding protein amplified in breast cancers, to exert its effect on enhancer assembly and dynamics. Overexpression of a targeted eRNA controlling expression of an oncogene increased cell proliferation, implicating eRNAs in regulating breast cancer cell growth. Together, we provide a genome-wide annotation of estrogen-regulated eRNAs, demonstrate the mechanisms by which targeted eRNAs promote target gene expression in vivo, identify the FERM element within eRNAs that is critical for regulating target gene expression, and show the biological implications of oncogenic eRNAs in breast cancer growth. Collectively, our results demonstrate the biological and mechanistic roles of eRNAs in breast cancer.

This work is supported by grants from the NIH/NIDDK (DK058110) and CPRIT (RP160319) to W.L.K.

Presentation: Saturday, June 11, 2022 11:30 a.m. - 11:45 a.m.