bluuebunny/crossref_metadata_2025 huggingface dataset:
SAT-200 Clinical Dilemmas of Adrenal Incidentaloma, Is Change in HU Clinically Significant?
Authors: Niaz Azra , Jun John , Ali Sophia Y
Published: May 2020

Introduction

There has been a recent increase in the incidence of adrenal incidentaloma (AI), defined as an adrenal mass that’s unintentionally discovered on imaging obtained for an indication other than suspected adrenal pathology, mainly because of increased availability and improved quality of imaging modalities.

Differentiation based on malignant vs benign and secretory vs non secretory becomes the most important question on finding such mass. Another important question is long term follow up of these AI. We present a case that highlights the importance of follow up due to the potential of AI to increase in size and density over 3 year.

Case

A 58 years old male with a history of ESRD secondary to hypertension, controlled hypertension, CAD s/p CABG was seen for a right 1.9 cm AI (found in 11/2016 on noncontrast CT abdomen) with low attenuation of approximately 10 Hounsfield units (HU).

He was asymptomatic, normal vitals, and an unremarkable physical exam. Hormonal work up was negative for hyperaldosteronism and Cushing syndrome.

About 1 year later, repeat CT abdomen reported an unchanged right, 2.0 x 1.7 cm adrenal mass, 22 HU. Hormonal work-up was not completed and patient was lost to follow up.

Two years later, repeat CT abdomen reported increase size of AI to 2.7 cm and increase in attenuation of 24 HU precontract which increased to 55 HU on the immediate postcontrast exam and 55 HU on 13-minute delayed images. Hormonal work up was negative for Cushing syndrome and hyperaldosteronism. Plasma metanephrine was 0.50 with elevated plasma normetanephrine of 3.85.

In light of being a renal transplant patient and due to increase in size, change in HU, and significantly elevated normetanephrine levels patient was referred for right adrenalectomy.

Discussion

Change in adrenal mass size is known to be a significant predictor of malignant potential. An unenhanced attenuation value <10 HU is characteristic of a lipid-rich adenoma, can predict benign adrenal adenoma with 98% specificity, and has a very low likelihood of a pheochromocytoma.

European Society of Endocrinology recommends if the AI is homogenous, < 4 cm, with a density ≤ 10 UH, no imaging follow up is recommended and biochemical testing for ruling out pheochromocytoma is indicated in adrenal tumors with an unenhanced attenuation value >10HU. However, the AACE/AAES guidelines recommends AI ≥ 1cm and less than 4cm, repeat imaging with noncontrast CT should be performed at 3–6 months and annually for 1-2 years and if the mass grows or becomes hormonally active then adrenalectomy should be performed.

Conclusion

There are no prospective studies of the optimal frequency and duration of follow up for AI. Considering the increasing incidence of AI, further studies on the clinical significance of change in HU, such as risk of developing pheochromocytoma, along with standardized international guidelines would be helpful for clinicians in managing patients with AI.
SUN-736 Knockout of Membrane Androgen Receptor ZIP9 Results in Reduced Female Fecundity and Abnormal Egg Activation in Zebrafish
Authors: Converse Aubrey , Thomas Peter
Published: May 2020

Recently, our research group cloned and characterized a putative membrane androgen receptor from teleost ovarian tissue that was found to be homologous with the zinc transporter protein ZIP9 (Slc39a9). To date, ZIP9 is the only zinc transporter that is known to be ligand activated or possess steroid receptor activity. Since the discovery of its androgen receptor activity, ZIP9 has been found to mediate androgen actions in a variety of tissues including teleost ovarian follicle cells, human cancer cell lines, and murine Sertoli cells. However, ZIP9 has not been examined in an in vivo model so the precise physiological functions of this receptor remain unclear. A ZIP9-mutant strain of zebrafish was developed using a CRISPR-Cas9 system in order to examine the role of the protein in teleost reproduction. While ZIP9-mutant males had similar breeding occurrence and fertilization rates to wild-type fish, mutant females exhibited severe reductions in fecundity compared to wild-type fish. ZIP9-mutant females spawn significantly fewer eggs of which a high proportion failed to undergo chorion elevation, a characteristic of normal egg activation. Eggs that showed this failed chorion elevation phenotype had significantly lower fertilization rates and produced larvae that exhibit a high incidence of pericardial/yolk sac edema and reduced growth compared to larvae hatched from wild-type eggs. However, no differences were observed in the proportions of oocytes at later stages of development between ZIP9-mutant and wild-type fish, suggesting the observed phenotypes are not related to abnormal oogenesis. We observed that mature wild-type eggs have numerous cortically located vesicles that are autofluorescent under ultraviolet light and decrease in number when the eggs undergo activation, suggesting they undergo exocytosis during the cortical reaction. While zinc is known to be stored in vesicles that undergo exocytosis in mammalian eggs, the role of zinc in teleost egg activation is currently unknown. In eggs from wild-type fish, we observed an increase in extracellular zinc levels upon egg activation and treatment with a zinc ionophore (zinc pyrithione) significantly reduced the number of eggs that undergo normal chorion elevation when activated. This suggests a role for zinc in zebrafish egg activation similar to that observed in mammals. Of interest, ZIP9-mutant eggs that did not undergo chorion elevation had significantly smaller vesicles than those found in wild-type fish eggs. This abnormal vesicle morphology and failure to undergo chorion elevation suggest a role of ZIP9 in egg activation. Additional insight into the role of zinc in zebrafish egg activation and the mechanism by which ZIP9 disruption leads to abnormal cortical vesicles and egg activation will help determine if ZIP9 plays a role in zinc transport and flux in zebrafish eggs during activation.
OR04-04 Identification of a Novel Transcriptional Regulator of Metabolic Disease in Circulating and Central Myeloid Cells
Authors: Sweet David R , Vasudevan Neelakantan T , Fan Liyan , Booth Chloe E , Keerthy Komal S , Jain Mukesh K
Published: May 2020

Derangement in systemic metabolic homeostasis is tightly associated with widespread activation of resident and circulating immune cells, a phenomenon known as ‘metaflammation’. Numerous studies have explored the role of tissue resident and circulating macrophages in contributing to metaflammation, obesity, and their sequelae; however, there is a dearth of information regarding targetable transcriptional regulators of the genesis and persistence of metabolic disease. Here, we identify myeloid Krüppel-like factor 2 (KLF2) as a novel regulator of metabolic disease. Previous reports demonstrate that KLF2 serves as a critical regulator of myeloid cell quiescence and is downregulated in numerous acute and chronic inflammatory states. Specifically in the context of chronic metaflammation, we note that KLF2 expression is decreased in circulating immune cells of obese patients and in adipose tissue macrophages of high fat diet (HFD) fed mice, which is consistent with the hypothesis that KLF2 regulates metaflammation. To explore this further, we utilized mice with myeloid cell-specific deletion of KLF2 (K2KO) which exhibit accelerated obesity and insulin resistance. K2KO mice have widespread central (i.e. CNS) and peripheral metaflammation both in the basal and HFD-stimulated states. To discern whether the effect of myeloid deletion of KLF2 on metabolism is due to deletion in microglia in the feeding centers of the hypothalamus or in peripheral immune cells, bone marrow chimeras with head shielding were created. 50% reconstitution of circulating immune cells with K2KO cells in wildtype (WT) mice was sufficient to maintain the metabolic disease phenotype, while mice with K2KO microglia + WT circulating cells had only slightly improved outcomes compared to K2KO mice. Conversely, ablation of microglia in K2KO mice using PLX5622 formulated in HFD also successfully attenuated the aberrant feeding behavior, weight gain, and glucose dyshomeostasis seen in K2KO mice. Together, these data demonstrate a role for loss of KLF2 in hematopoietic and CNS resident cells in causing metabolic disease. Given that myeloid KLF2 expression decreases under metabolic stress in WT mice and humans, we sought to explore whether maintenance of KLF2 expression in these cells would be protective against diet-induced metabolic disease. Indeed, mice with myeloid-specific overexpression of KLF2 demonstrated a markedly improved metabolic phenotype when challenged with HFD, providing evidence that targeting KLF2 expression in myeloid cells may prove to be a therapeutic option against metaflammation.
MON-588 Anthropometric Parameters, Body Fat Percentage and Metabolic Profile in Sarcopenic Women with Recommendation for Bariatric Surgery
Authors: Carvalho Nara Nóbrega Crispim , Limeira Caio Chaves de Holanda , Borges Heloisa Calegari , Lima Débora Nóbrega , Mesquita Patricia Nunes , Rego Gitana Silva , Holanda Narriane Chaves Pereira , Martins Vinícius José Baccin , Filho João Modesto , Pimenta Flávia Cristina Fernandes , Alves José Luiz de Brito
Published: May 2020

INTRODUCTION: Sarcopenia (SARC) is a musculoskeletal disorder that predisposes several complications, including metabolic ones. Obesity also provides higher risk for metabolic complications, however, there is lack of evidences regarding the association of obesity with SARC on metabolic parameters in non-elderly individuals. OBJECTIVE: To evaluate anthropometric parameters, body fat percentage (BFP) and metabolic parameters in women with and without SARC preceding Bariatric Surgery (BS). METHODS: A cross-sectional study involving 60 obese women in the outpatient care in a public Brazilian University Hospital between March to September 2018. Body composition was given by bio-impedance (inbody-370), multifrequency (5, 50, 250Hz) with 12 hours fasting, dominant Handgrip Strength (HS) was evaluated by Jamar dynamometer (3 measurements; 30 sec interval). Were also evaluated fasting blood glucose, HbA1c, homeostatic model assessment-insulin resistance (HOMA-IR), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), triglycerides and high-sensitive C-reactive protein (hs-CRP). SARC was defined by the association of a low muscle mass index (weight-adjusted appendicular skeletal muscle mass: ASMM/weight x 100%) and decreased HS, using as cutoff points the smallest quintile for each variable. Data were expressed as mean ± standard deviation and independent t-test was used for comparison between groups. Statistics were made by SPSS software, 20th version (IBM Corp., Armonk, NY). RESULTS: The mean age, weight, body mass index and BFP of sarcopenic and non-sarcopenic women were: 40.75 ± 11 x 39.23 ± 8.92 years old (p=0.665), 102.93 ± 9.58 x 109.19 ± 14.25 Kg (p=0.237), 44.88 ± 2.7 x 42.24 ± 4.79 Kg/m2 and 54.12 ± 1.11 x 51.44 ± 3.43% (p=0.052), respectively. Regarding the laboratory parameters of women with and without SARC: fasting blood glucose 89.25 ± 14.48 x 98.40 ± 27.48 mg/dL (p=0.359), HbA1c 5.83 ± 0.33 x 6.21 ± 1.18% (p=0.185), HOMA-IR 3.61 ± 1.28 x 5.31 ± 4.74 (p=0.160), TC 170.87 ± 39.36 x 180.82 ± 34.51 mg/dL, LDL 94.67 ± 26.63 x 105.60 ± 30.85 mg/dL, HDL 53.37 ± 18.50 x 50.84 ± 10.32 mg/dL, triglycerides 114.12 ± 38.84 x 127.30 ± 75.04 mg/dL and hs-CRP 8.51 ± 6.50 x 7.51 ±6.52 mg/L (p=0.792). CONCLUSION: Women with SARC and recommendation for BS when compared to women without SARC had similar anthropometric, metabolic and BFP parameters.
OR19-06 Sepsis-Induced Critical Illness in Mice Alters Key Regulators of ACTH Production and Secretion Within the Anterior Pituitary Gland
Authors: Téblick Arno , Perre Sarah Vander , Langouche Lies , Van den Berghe Greet
Published: May 2020

Introduction

Critical illness is hallmarked by high plasma cortisol without elevated ACTH. This is partly explained by low cortisol-binding proteins and reduced binding affinity and by suppressed cortisol metabolism, 3 peripheral drivers of increased free cortisol (corticosterone (CORT) in rodents).1 We hypothesized that lack of elevated plasma ACTH is explained by CORT-induced feed-back inhibition either due to suppressed CRH or AVP pituitary signaling or to direct glucocorticoid receptor (GR)-mediated action within corticotrophs. To test this hypothesis, we documented alterations in ACTH precursors and key enzymes controlling ACTH maturation and secretion in a mouse model of sepsis-induced critical illness.

Methods

C57Bl/6 mice were randomly allocated to a healthy control group or to 4 critically ill groups sacrificed after increasing illness duration (30 hours (H), 3 days (D), 5D or 7D). Critical illness was induced by sepsis brought about by cecal-ligation and puncture followed by fluid-resuscitation and antibiotics treatment. The study was continued until 15 surviving animals per time cohort were reached (n=120).

We quantified pituitary pro-opiomelanocortin (POMC) gene/protein expression and POMC plasma concentrations, pituitary POMC intracellular trafficking and cleavage via intracellular POMC sorting/trafficking receptor Carboxypeptidase E (CPE) and prohormone convertase 1 (PC1/3) gene/protein expression. Gene expression of Annexin A1, an inhibitor of mature ACTH secretion, was quantified as marker of GR-mediated CORT-induced feedback inhibition at corticotroph level.

Results

Plasma CORT concentrations were median 3-fold increased during critical illness (p<0.001 for all time cohorts) in the face of normal (for 30H, 3D and 5D cohorts) to low (7D time cohort; p=0.01) plasma ACTH concentrations. Plasma POMC concentrations were higher in critically ill than in control mice (p=0.05). POMC gene expression (but not protein, P=0.8) was a median 55% higher in critically ill mice than in controls (p<0.05 for all time cohorts). In contrast, pituitary mature ACTH protein concentration was median 61% lower in critically ill than in control mice (p<0.01). CPE gene expression was only increased in 30H time cohort (p<0.001). PC1/3 gene and protein expression were positively correlated (R2 0.1; p=0.001) and were reduced (by 37% and 43%, respectively) during the entire course of critical illness (p<0.01). Annexin A1 gene expression was increased during critical illness (p<0.05 for all time cohorts).

Conclusion

Suppressed CRH or AVP signaling and GR-mediated action within corticotrophs explained lack of elevated plasma ACTH in critical illness, as indicated by impaired POMC processing and ACTH maturation. However, increased POMC gene expression suggests ongoing corticotroph activation, the driver of which needs to be identified.

1. Teblick A et al. Nat Rev Endocrinol 2019
MON-018 Fertility Rates in Rats Characterized by Increased Hypothalamic CRH Secretion
Authors: Zoumakis Emmanouil
Published: May 2020

Certain strains of rats are characterized by hyperactive Hypothalamic-Pituitary-Adrenal axis responses to stress, increased hypothalamic Corticotropin-Releasing Hormone (CRH) production and decreased fertility rates. Activation of the HPA-axis and CRH secretion has been associated with suppression of the Hypothalamic-Pituitary-Ovarian axis primarily as a result of glucocorticoids. Here we examined the hypothesis that Fischer rats have decreased fertility rates because of hypothalamic CRH hypersecretion. Antalarmin, a CRH receptor type 1 antagonist, is known to suppress adrenocorticotropin hormone secretion and other CRH receptor type 1-mediated responses. Adult female Fischer rats were injected with antalarmin or placebo, twice a day, for 16 days. Mating was evidenced by the presence of spermatozoa in the vaginal smear performed every morning. After 16 days, 20% of rats (20%) treated with placebo became pregnant and 55% rats treated with antalarmin became pregnant. We have previously reported that administration of antalarmin after the first day of pregnancy does not affect blastocyst implantation in Fischer rats. Our data suggest that antalarmin improves fertility rates in Fischer rats by antagonizing the direct antireproductive role of hypothalamic CRH.
SAT-032 Follistatin-Like 3 (FSTL3) and Early Postnatal Testicular Development
Authors: Mejia Randy Ballesteros , Xia Dong , O’Shaughnessy Peter , Mukherjee Abir
Published: May 2020

Activin, a TGFβ family ligand, induces Sertoli cell (SC) proliferation during the early postnatal stage in mice. Overexpression of activin, however, leads to the disruption of spermatogenesis. FSTL3, a glycoprotein inhibitor of activin, is highly expressed in the testis and its expression is induced by activin creating an inhbitory feedback loop. FSTL3 deletion in mice is therefore expected to produce a mouse model of increased activin action. Contrary to overexpression of activin, however, as we have shown before, global deletion of FSTL3 in mice results in increased numbers of SC and increased sperm production in older males. Stereological analyses show that although the overall number of SC increases with age in both genotypes during pre-pubertal stages, increase in SC numbers is significantly higher in FSTL3 KO males. Here we show our transcriptomic analyses of WT and FSTL3 KO mice at 3d and 8wk. mRNA sequencing data showed that more than 1000 genes are diffferentially regulated between WT and FSTL3 KO at 3d. There is a much lower number of genes differentially expressed at 8wk. Among several canonical pathways that are altered at 3d in FSTL3 KO mice compared to WT we investigated the “Sertoli-to-Sertoli cell Communication” pathway. We found increased expression of junction proteins, including those that are involved in the blood testis barrier (BTB) as well as earlier establishment of BTB in FSTL3 KO testis. Without the BTB, preleptotene spermatocytes cannot progress through the spermatogenic programme. Importantly, we found accelerated SYCP3 organisation, indicative of early entry into meiosis, concomitant with early establishment of BTB. To directly address whether FSTL3 deletion can induce SC proliferation we knocked down FSTL3 expression in the TM4 SC line. We found increased PCNA expression with FSTL3 siRNA compared to control siRNA transfection demonstrating FSTL3 impairment does induce SC proliferation. We are currently investigating the mechanisms for early establishment of BTB in FSTL3 KO testes and additional canonical pathways. The results of the study presented suggests that loss of FSTL3 promotes SC proliferation which likely allows establishment of SC-SC interactions including BTB formation earlier than in WT. Given that the difference in size between WT and FSTL3 KO testis persists at older ages, it is possible that the timing of increase in SC number and establishment of BTB has a sustained effect on testicular function later in life.
SUN-733 Analysis of Divergent Long Noncoding RNAs in Estrogen-Regulated Transcription
Authors: Lee Debra , Yang Barabara , Sedano Melina , Choudhari Ramesh , Gadad Shrikanth S
Published: May 2020

The role of long noncoding RNAs (lncRNAs) in cancer biology are just beginning to be elucidated and recent studies have shown that they could be therapeutic targets. In a previous study, combining powerful techniques, Global Run-On sequencing (GRO-seq) and subcellular fractionation RNA-seq in breast cancer cells identified a large number of estrogen-regulated unannotated long noncoding RNAs. Analysis of gene expression data from hundreds of samples representing 13 different tissue types including both cancer and normal tissue, revealed that many lncRNAs are differentially expressed in various cancers. Furthermore, a large number of lncRNAs are divergent transcripts and show distinct expression patterns across molecular subtypes of cancer. In functional assays, knockdown of selected lncRNA, such as lncRNA67, inhibits the growth of breast cancer cells. Amplified expression of lncRNA67 in luminal-subtype of breast cancer correlates with clinical outcome. LncRNA67 has now been fully annotated (transcription start and stop site, 5’ cap, polyA tail, and exon/intron structure), and cloned. Our preliminary molecular analyses indicate that lncRNA67 plays a critical role in ER-dependent and -independent pathways. Collectively, our results suggest that lncRNAs are an integral component of cancer biology.
OR04-02 The Role of the Sympathetic Nervous System in Metabolic Disorder and Adipose Dysfunction in Obesity and Aging
Authors: Sakamoto Kenichi , Zhou Chunxue , Maurizi Giulia , Liberini Claudia , Li Ling , Ruiz Henry H , Buettner Christoph
Published: May 2020

Both obesity and aging increase susceptibility to metabolic disease and type 2 diabetes where adipose tissue dysfunction is a hallmark. In both conditions, impaired autonomic control is believed to play a key role in disrupted CNS control of metabolism. However, the role of the sympathetic nervous system (SNS) and catecholaminergic signaling in metabolic disease has not been well defined in large part due to a lack of suitable animal models(1). Surgical denervation is not specific to the SNS and chemical sympathectomy through 6-hydroxydopamine causes inflammation due to toxic effects. Abrogation of catecholamine (CA) synthesis in the whole body due to genetic deletion of the gene for tyrosine hydroxylase (th), a key enzyme in CA synthesis, results in embryonic lethality likely due to the lack of dopamine and norepinephrine in the CNS where they serve as key neurotransmitters.

Here we studied the role of the SNS and catecholaminergic signaling in metabolic control in both aging as well as high fat diet (HFD) induced obesity. We created a mouse model of inducible th gene deletion that is restricted to the periphery, including sympathetic fibers of the peripheral NS but spares the brain as a pharmaco-genetic model of sympathectomy(2). TH is deleted and CA levels were reduced more than 90% in peripheral tissues of TH KO mice, while intact in the CNS. TH KO mice are cold intolerant consistent with functional sympathectomy. Interestingly, TH KO mice are protected from HFD feeding induced glucose intolerance (AUC during GTT: WT1018.8±42.0 mg/dl/hr vs. TH KO 485.0±85.8 mg/dl/hr; p < 0.0001; n = 6) even though food intake increased in TH KO mice. In 20 months old TH KO mice glucose tolerance was improved and fasting blood glucose levels were reduced (AUC during GTT: WT 357.3±16.2 mg/dl/hr vs. TH KO 254.5±15.6 mg/dl/hr; p < 0.01; n = 12) with higher insulin levels (WT 0.35±0.07 μg/l vs. TH KO 1.28±0.28 μg/l; p < 0.001; n = 9). Of note, insulin tolerance tests did not show marked differences. Both obesity and aging are characterized by impaired adipose tissue function with reduced lipogenic capacity. TH KO mice fed a HFD exhibit increased WAT de novo lipogenesis, lower lipolysis, and trend to exhibit decreased adipose tissue inflammation, suggesting that the SNS is a major culprit for the impaired lipogenic capacity in adipose tissue. Our data provides support for the paradigm that impaired SNS function plays an important role in the dysmetabolic states of obesity and aging.

Reference

1. Ryu V, Buettner C. Fat cells gobbling up norepinephrine? PLoS Biol. 2019;17(2):e3000138.

2. Fischer K, Ruiz HH, Jhun K, Finan B, Oberlin DJ, van der Heide V, et al. Alternatively activated macrophages do not synthesize catecholamines or contribute to adipose tissue adaptive thermogenesis. Nature medicine. 2017;23(5):623-30.
SUN-592 Adipocyte Specific Endothelin a Receptor Knockout Increases Adiposity in Mice
Authors: Rivera-Gonzalez Osvaldo , Speed Joshua S
Published: May 2020

Obesity is associated with increased levels of Endothelin-1 (ET-1). Blockade of ET-1 type A receptors (ETA) improves lipid profile in patients with chronic kidney disease; however the mechanism is unknown.[1] In adipocytes ETA activation increases lipolysis, a potential mechanism for elevated lipids in obese individuals.[2] Therefore, the goal of this study was to determine if adipocyte specific knockout (KO) of the ETA receptor in mice alters genes associated with lipid metabolism in adipose and improves plasma lipids. 24-week old adipocyte ETA knockout mice had significantly elevated body weight compared to floxed controls (32.6±1.0 vs. 29.5±0.7 g respectively). Echo MRI revealed that the increased body weight was due to greater adiposity (10.1±0.9 vs. 14.7±1.8 % body weight; floxed vs. KO), while no statistical difference was observed in lean weight (88.9±2.4 vs. 86.8±2.6 % body weight; floxed vs. KO). Surprisingly, there were no statistical differences in plasma total cholesterol or triglycerides. RNA sequencing indicated downregulation of 597 genes and upregulation of 444 genes in visceral adipose and downregulation of 368 and upregulation of 847 genes in subcutaneous adipose. KEGG pathway analysis revealed that most genes altered in visceral adipose were related to metabolic pathways. These data implicate a role for adipose tissue ETA receptors in regulating adiposity and promoting pathophysiology related to obesity.

1.

Farrah, T.E., et al., Endothelin Receptor Antagonism Improves Lipid Profiles and Lowers PCSK9 (Proprotein Convertase Subtilisin/Kexin Type 9) in Patients With Chronic Kidney Disease. Hypertension, 2019. 74(2): p. 323-330.

2.

Eriksson, A.K., et al., Endothelin-1 stimulates human adipocyte lipolysis through the ET A receptor. Int J Obes (Lond), 2009. 33(1): p. 67-74.
SAT-435 A Direct Comparison of Thyroid Hormone Receptor (THR) Protein Levels in Mice Provides Unexpected Insights into TH Action
Authors: Bansal Sanya , Minakhina Svetlana , Zhang Alice , Brotherton Michael , Janodia Rucha , Oliveira Vanessa De , Tadepalli Shaan , Wondisford Fredric Edward
Published: May 2020

Thyroid Hormone (TH) action is mediated by three major THR isoforms α1, β1 and β2 (THRA1, THRB1 and THRB2), encoded by two genes Thra and Thrb. A fourth non-T3 binding isoform is termed THRA2. Previous characterization and comparison of THR isoform expression using QPCR of mRNA levels does not assess translational or post-translational control of THR protein levels. Moreover, reliable antibodies against all THR isoforms are not currently available. To address these concerns, we generated knock-in mouse models expressing endogenously and identically 2X HA tagged THRs (THRA1/2, THRB1 and THRB2), which could then be detected by commercially available anti-HA antibodies. We characterized THR expression in 16 mouse organs. We found that in all peripheral tissues tested except the liver, the dominant THR isoform is THRA1, and THRB1 and THRA2 are expressed at significantly lower levels or are undetectable. Surprising THRB1 levels were very low in metabolically active organs such fat and muscle. In some organs, mRNA transcript levels predicted THR protein amounts, while in others the prediction was inaccurate. For instance, adipose depots have similar levels of Thrb1 and Thra1 transcripts, however THRA1 protein levels are up to 10-fold higher than THRB1. In contrast to peripheral organs, brain tissues express low levels of both THRB1 and THRA1, but have very high levels of THRA2. As expected and confirmed in this study, THRB2 has limited expression and was only detected in pituitary of the organs tested. Interestingly, THRB2 expression in female was much higher than male mice (the only sex-difference in THR expression found); and expression of the THRB2 target gene, Tshb, was lower in female mice. For the first time, these HA-THR mouse lines can be used to accurately compare isoform-specific actions of THRs in organs. The predominant expression of THRA1 in most peripheral organs, for example, suggests that many peripheral actions of THRB1 could be indirectly mediated.
MON-335 Bisphosphonate and Denosumab Refractory Hypercalcemia of Malignancy: What Else Is at Play?
Authors: Patel Reshma , Lopresti Jonathan S
Published: May 2020

Hypercalcemia of Malignancy has been historically responsive to anti-resorptive agents. However, when multiple mechanisms contribute, it may be difficult to treat with one modality. This case highlights the importance of the work up in treatment of hypercalcemia in a low PTH state. A 44 yo M with h/o high grade metastatic spindle cell neoplasm with skeletal metastasis was admitted with hypercalcemia. He reported some constipation prior to presentation, however denied confusion. His vital signs were notable for HR of 86 bpm and BP of 112/75 mmHg. Labs at admission were remarkable for an uncorrected Ca of 16.1 (8.8-10.3 ng/mL), a phosphorus (Phos) level of 3.3 mg/dL (2.5-4.5 mg/dL), a PTH level of 11 pg/mL (15-65 pg/mL), PTHrP level of 134 pg/mL (14-27 pg/mL), a 25 OH vit D level of 11 ng/mL (30-100 ng/mL), and a BUN/Cr and GFR of 34/2.38 (8-22 mg/dL/0.5-1.3 mg/dL) and 32 ml/min/m2. He was given intranasal calcitonin and ergocalciferol, then received 2mg IV of zolendronic acid, which reduced the patient’s Ca level to a nadir of 6.6 ng/mL in 5 days. On the next admission serum Ca was elevated to 15.7 ng/dL, which did not respond to zolendronic acid. Given that patient’s Ca was refractory to zolendronic acid, denosumab was given but had no response. He then underwent surgery for cord compression and was given dexamethasone (dex) 4mg IV Q6h post-op. His Ca responded quickly to dex, with a nadir to 9.2 ng/dL, however his Ca became elevated after cessation. Given response to dex, vit D 1,25 OH level was sent and was elevated at 94 pg/mL (18-72 pg/mL). In addition, given his inappropriately normal Phos level in the setting of low PTH, FGF23 was sent and came back elevated at 473 RU/mL (<180 RU/mL). This was likely due to increased bone turnover and release of FGF23. He was discharged with a Ca level of 12.5 ng/mL, however was found to have an elevated Ca to 14.9 ng/mL on presentation to clinic. Given concern that Vit D 1,25 OH, PTHrP and direct bony involvement were all contributing to his hypercalcemia, patient was started on IVF and dex IV. His calcium responded 11.5 ng/mL and was then transitioned to PO dexamethasone and plaquenil. The most likely explanation for this phenomenon is malignancy induced cytokine/PAMP release, which stimulates 1-alpha hydroxylase in tumor macrophages to convert 25 OH D to 1,25 OH D. This was supported by his elevated 1,25 OH D level and a decreased 25 OH D, which suggests that 25 OH D was used as substrate by activated macrophages. This case highlights the importance of ancillary work up of hypercalcemia when a patient’s calcium is refractory to standard anti-resorptive therapy. Moreover, it shows the need for a systematic approach when treating hypercalcemia.
MON-648 Chronic Unpredictable Environmental Stress May Induce Predisposition to Diabetes Mellitus
Authors: Raghav Alok
Published: May 2020

Title: Chronic Unpredictable Environmental Stress may induce predisposition to diabetes mellitus Objective: Chronic unpredictable environmental stress (CUES) may induce predisposition to diabetes mellitus. Material & Methods: This study investigates the role of CUES on impaired homeostasis. Stressed group mice (n=20) were exposed to CUES for 16 weeks. Weekly body weight, feed consumption, feed efficiency ratio, fasting blood glucose were monitored. Plasma HbA1c, plasma cortisol, plasma epinephrine and plasma insulin, serum lipids, antioxidants and carbohydrate metabolizing enzymes activity were assessed along with DNA damage and histopathological examination of liver, kidney, pancreas, spleen and skeletal muscles. Semi-quantitative expression of IL-4, IL-6 and β- actin was also assessed. Results: Fasting blood glucose levels & HbA1c in the stressed were significantly higher compared to control (p<0.001). Serum lipids were found insignificantly higher in stressed mice compared to control. Body weights of the stressed mice and feed efficiency ratio were found significant (p<0.001). Plasma corticosterone, plasma epinephrine, HOMA-IR was found to be significantly higher in the stressed group (p<0.001). Plasma insulin level was found to be significantly lower in the stressed group (p< 0.001). Significant changes were observed in antioxidants level, carbohydrate metabolizing enzymes activity, peripheral tissues and DNA integrity. Expression of IL-4, IL-6 was found significantly higher in the stressed group. Conclusions: CUES initiates pathogenesis of diabetes.
OR03-02 Identification of a Novel Stem/Progenitor Population of the Adrenal Medulla
Authors: Santambrogio Alice , Russell John P , Lodge Emily J , Scriba Laura D , Berger Ilona , Hakansson Hanna , Steenblock Charlotte , Eisenhofer Graeme , Fassnacht Martin , Bornstein Stefan R , Andoniadou Cynthia L
Published: May 2020

The adrenal glands regulate multiple physiological processes including the stress response, the immune system and metabolism. The adrenal is composed of an outer cortex that produces steroids, and an inner medulla that produces catecholamines. Tissue-specific stem/progenitor populations have been identified in the adrenal cortex, while the presence of a functional stem/progenitor population in the adrenal medulla is unclear. The adrenal medulla derives from the neural crest and contains chromaffin cells, neurons and sustentacular (support) cells. Establishing cell hierarchy and elucidating mechanisms of regulation of the different cell types is important to understand normal homeostasis and disease pathogenesis, such as of pheochromocytomas. Using genetic approaches in mouse, we have established that a subpopulation of sustentacular cells express the stem/progenitor marker SOX2. Through genetic lineage-tracing using the Sox2-CreERT2 strain, we demonstrate that these are an expanding population, capable of giving rise to chromaffin cells and neurons throughout life, consistent with a stem/progenitor role in vivo. We further demonstrate the self-renewal and differentiation potential of SOX2+ cells through in vitro isolation and expansion. Through analysis of FFPE sections of human adrenals, we confirm the presence of SOX2+ cells in the normal adult organ, as well as in pheochromocytomas. Taken together, our data support the identification of a previously undescribed stem/progenitor cell in the mammalian adrenal medulla, and confirm its functional relevance.
MON-251 Xanthogranulomatous Hypophysitis: A Rare Case of Hypopoituitarism
Authors: Romero Fabiola , Lopez Helen , Terwindt Natalia , Sanchez Humberto , Ocampos Sara , Avalos Andres , Boggino Hugo , Pfannl Rolf , Ayala Alejando
Published: May 2020

The patient is a 54-year-old male with type 2 diabetic treated with insulin who presented with a two-week history of fever nausea and vomiting. He subsequently developed seizure episodes with changes in mental status prompting a visit to the emergency room. Upon admission the patient was hypotensive and lethargic. After an initial improvement, he developed polyuria (14-15 liters per day) in the context of eunatremia and specific gravity of 1010. His initial evaluation revealed a serum a.m. cortisol of 1.2 mcg/dL (6.2 to 19.4), low free T4 of 0.64 Ng/dl) 0.9-1.7) and secondary hypogonadism with a total testosterone of less than 2.5 ng/dL (181-772). Hormonal replacement was initiated. MRI of the brain revealed heterogeneous sellar lesion measuring 2.1 x 2.0 x 2.0 cm with peripheral enhancement following contrast administration and deviation of the pituitary stalk to the right. The lesion invaded the left cavernous sinus (Knosp III) and abutted the optic chiasm. He underwent an uneventful transsphenoidal tumorectomy revealing extensive coagulation necrosis with chronic lymphocytic infiltration, numerous macrophages and multinucleated giant cells with deposits of hemosiderin, cellular debris and isolated cholesterol crystals surrounded by granulomatous tissue. The pathology was compatible with xanthogranulomatous hypophysitis. We report a rare case of Xanthogranulomatous hypophysitis (XGH), the least common subtype of pituitary hypophysitis often mistaken by a neoplastic lesion. It is more common in young adults occurring in isolation, as a secondary reaction to a local process such as apoplexy part or as part of a systemic condition with autoimmune aetiology, often resulting in variable degrees of anterior and posterior pituitary deficiencies.
SUN-660 Appetite Suppressing Effects of Glucoregulatory Chimeric Peptides Devoid of Nausea
Authors: Roth Christian Ludwig , Milliken Brandon T , Elfers Clinton , Sweet Ian , Doyle Robert
Published: May 2020

Current treatments for type 2 diabetes (T2D) and obesity do not reliably achieve long-term weight-loss and up to 50% of patients experience nausea and vomiting. Thus, there is a critical need for obesity medications that provide glycemic control with enhanced hypophagic response without nausea. Based on rational design and in silico modelling of the glucagon-like peptide 1-receptor agonist (GLP1RA) exendin-4 (Ex4) and the neuropeptide Y2 receptor (Y2R) agonist peptide YY (PYY) 3-36, our group has developed and tested several new monomeric chimeric peptides that display dual agonism of the anorectic Y2R and the glucoregulatory GLP1R. In addition, we explored a third agonistic behavior at the neuropeptide Y1 receptor (Y1R), which potentially relates to pancreatic beta-cell protection. Our current lead, GEP44, binds and robustly activates the Y2R, Y1R, and GLP1R. In our experiments, half maximal effective concentration (EC50) values for GEP44 were 10 nM at Y2R, 24 nM at Y1R, and 330 pM at GLP1R for dose responses of cell-based fluorescence resonance energy transfer assays. For the control substrates PYY3-36, PYY1-36, and Ex4, EC50 values were 16 nM, 12 nM, and 16 pM at the Y2R, Y1R, and GLP1R, respectively. We tested effects of daily injections of these chimeric peptides in adult Sprague-Dawley rats on food intake (FI), body weight (BW) changes, blood glucose levels and an indicator of nausea (kaolin intake). GEP44 produced profound reduction in FI (2-d average GEP44 at 20 nmol/kg FI -71%; Ex-4 20 nmol/kg FI -40%). Anorectic doses of GEP44 did not trigger a pica response assessed by kaolin consumption in treated rats, while in Ex-4 treated rats, kaolin consumption accounted for 28% of total daily solid intake, indicating a clear nausea response. During 11 d of treatment with GEP44, FI was consistently reduced resulting in a significantly stronger reduction of BW compared to Ex4 at the end of treatment (GEP44 -7.6%, Ex4 -3.7%, p<0.05). Furthermore, 5-day injections of GEP44 showed greater reduction of glucose levels than for Ex4 in diet induced obese rats receiving intraperitoneal glucose. These results were supported by results generated using a customized perifusion system that demonstrated acute stimulation of insulin secretion by exposure of isolated rodent and human islets to GEP44 and Ex4 (both p<0.01 vs. vehicle). In conclusion, simultaneously targeting serial anorectic pathways with single molecule chimeric peptides developed by our group can address multiple coexisting conditions, namely obesity and T2D, to more efficaciously reduce FI, BW and blood glucose levels thereby reducing off target effects.
SUN-211 Active Steroid Hormone Synthesis Renders Adrenocortical Cells Highly Susceptible to Type II Ferroptosis Induction
Authors: Weigand Isabel , Schreiner Jochen , Roehrig Florian , Sun Na , Laura-Sophie Landwehr , Urlaub Hanna , Kiseljak-Vassiliades Katja , Wierman Margaret E , Angeli José Pedro Friedmann , Walch Axel K , Sbiera Silviu , Fassnacht Martin , Kroiss Matthias
Published: May 2020

Context: Cell death in the adrenal cortex is ill understood but of high clinical relevance. Resistance of adrenocortical carcinoma (ACC) to current treatment with mitotane and chemotherapy calls for an improved understanding of adrenal cortical cell death processes. Ferroptosis is an iron-dependent form of regulated cell death which is characterized by polyunsaturated lipids adrenic (AdA) and arachidonic acid (AA) peroxidation. Aim: To address the potential role of ferroptosis in the adrenal gland as a potential treatment target of ACC. Methods: Human ACC cells H295R, CU-ACC1 and 2 were used. Protein expression of key enzymes was determined by western blotting. Lipid peroxidation was quantified with BODIPY 581/591 and cell viability with CellTiterGlo after treatment with known inducers and inhibitors of ferroptosis and steroidogenesis, respectively. Results: Adrenocortical tissues are enriched in AdA and AA and express high levels of genes relevant to ferroptosis, such as glutathione peroxidase 4 (GPX4) and long-chain-fatty-acid CoA ligase 4 (ACSL4). Inhibition of GPX4 with RSL3 led to cell death in H295R, CU-ACC1 and 2 cells at EC50 values of 2.4x10-7, 8.1x10-7 and 1.5x10-8 M, respectively. The steroidogenesis inhibitor ketoconazole completely reversed RSL3 cytotoxicity in all three steroidogenic cell lines by reducing lipid peroxidation. Mitotane induced lipid peroxidation but inhibition of ferroptosis with liproxstatin did not protect mitotane-induced cell death. Conclusion: Adrenocortical cells are highly sensitive to ferroptosis due to active steroidogenesis. Triggering this form of cell death could present future novel treatment options against ACC.
SUN-484 Complex Thyroid Cancer with Several Recurrences and Metastases
Authors: Patel Kruti K , Via Michael
Published: May 2020

56-year-old male with metastatic papillary thyroid cancer who underwent total thyroidectomy in 8/2014, and I-131 ablation in 9/2014, with post-operative pathology revealing multifocal, bilateral papillary thyroid cancer with extrathyroidal extension with surgical resection margins uninvolved. There were positive lymphovascular spread and multiple central compartments and bilateral neck nodes metastases with extranodal extension. Subsequent thyrogen stimulated whole-body scan in 2/2016 showed no areas of uptake.However, in 12/2016 he was found to have right supraclavicular lymph nodes positive for recurrence which was resected and given another 168 mCi I-131 and 33 treatments of XRT to R shoulder.A repeat PET in 7/2019 showed persistent hyper metabolic lesion in C7, multiple hyper metabolic nodules throughout the R lung, and a new 1.1 cm left Hilar lymph node suggesting disease progression. Biopsy of the C7 lesion confirmed dedifferentiated papillary thyroid cancer and demonstrated the presence of an NTRK mutation.This time he was given lenvatinib 24 mg daily for 4 weeks, followed by 200 mCi I-131. Post-treatment whole body scan showed good uptake in all lesions, except the C7 lesion which was treated with external radiation. DiscussionWhile cure is achieved in most cases of differentiated thyroid cancer, a minority of cases demonstrate disease progression. Loss of response to I-131, very low serum thyroglobulin levels despite known disease, and high PET avidity provide clinical evidence of dedifferentiation, confirmed with tissue sampling.If feasible targeted systemic therapy remains the best tolerated treatment option.While several studies demonstrate an increase in iodine avidity in approximately 50-60% of patients with dedifferentiated thyroid cancer that were treated with tyrosine kinase inhibitors (TKI), (cite alan Ho’s 2013 NEJM article, and the 2015 debrafenib study Rothenberg SM et al, clin cancer res 2015), selumetanib remains unavailable for clinical use and dabrafenib may only be beneficial in cases with known BRAF V600E mutations. Moreover, it is unknown whether a planned short course of TKI therapy would potentially induce resistance to future TKI therapy.Therefore, lenvatinib, which inhibits activity of at least 6[VM1] different tyrosine kinase enzymes important in thyroid cancer was chosen rather than entrectanib, which was reserved for use if the need arises. This patient demonstrated excellent response to I-131 therapy with lenvatinib pretreatment.A number of formal studies of various TKIs for thyroid cancer re-differentiation are currently underway. (cite Brown SR, Hall A, et al BMC cancer 2019; and also cite the CIII trial with cabozatanib)Conclusion This case represents the emerging paradigm for the ability of TKI therapy to redifferentiate advanced thyroid cancer and allow for re-treatment with I-131 targeted therapy.
SUN-638 Perinatal DDE Exposure Disrupts Thermogenesis Early in Development
Authors: vonderEmbse Annalise , Tong Jason W , Lein Pamela J , Merrill Michele La
Published: May 2020

Numerous epidemiological studies have identified a positive association of exposure to the endocrine-disrupting pesticide dichlorodiphenyltrichloroethane (DDT) and its primary metabolite dichlorodiphenyldichloroethylene (DDE) with the risk of obesity. Of particular concern is the persistent metabolic dysfunction resulting from early life DDT and DDE exposures, evidenced as obesity, glucose intolerance, dyslipidemia, and impaired thermogenesis in adult rodents. However, little is known about the developmental timing and etiopathogenesis of this long-term DDT-related metabolic phenotype. This study aimed to address these knowledge gaps by evaluating endocrine phenotypes and thermogenic parameters at two postnatal (P) time points, P0 and P12, in C57BL/6J mice. Dams were orally gavaged with p,p’-DDT (1.7 mg/kg body weight [BW]), p,p’-DDE (1.31 mg/kg BW), or organic olive oil (vehicle) daily from gestational day (GD) 11.5-P0 or P5. Infrared analysis was then performed during a cold challenge. We further attempted to rescue the cold-challenged P12 mice by administering the β3-adrenergic receptor (AR)-agonist CL316,243 (CL), a direct stimulator of brown adipose (BAT) thermogenesis. At P0, area under the curve analysis revealed higher body temperatures in both males and females exposed to DDE compared to controls during the 9 min cold challenge. In addition, DDE-exposed females lost body heat at a significantly faster rate than sex-matched controls over the 9 min cold challenge. This occurred without any treatment-related differences in resting glucose, suprascapular BAT weight, or body weight for either sex at P0. To assess BAT-autonomous response to β3-AR stimulation during a cold challenge, P12 mice were exposed to the pharmacological β3-AR agonist, or PBS control in a 2x2 exposure design and suprascapular BAT temperature was evaluated via infrared analysis of the suprascapular surface at 10 min intervals. Blood glucose, BAT weight, and BW remained equivocal across all P12 treatment groups. However, BAT temperature was significantly decreased 10 min after cold- challenge in female P12 mice with perinatal DDE-exposure when compared to sex- and perinatal treatment-matched controls. This DDE effect among P12 female mice was rescued by CL. These data suggest that thermogenic dysfunction consequent to perinatal DDE exposure is detectable during postnatal development, well before the emergence of endocrine phenotypes. The CL rescue of the BAT thermogenic impairments observed after perinatal DDE exposure are consistent with DDE toxicities upstream in the β3-AR nerve circuitry. Additionally, the data highlight the emergence of an early postnatal sex divergence in DDE-related thermogenic dysfunction. We speculate that the effects of DDE on suprascapular BAT heat production result from prenatal alterations in sympathetic circuitry.