bluuebunny/crossref_metadata_2025_split huggingface dataset:
MON-726 Modifications of FOXO1 and GATA4-NKX2.5 Signaling Induce Human Enteroendocrine Differentiation
Authors: Zeve Daniel Richard , Stas Eric , Shah Manasvi S , Breault David T 
Published: May 2020

Abstract
Enteroendocrine (EE) cells are the most abundant hormone-producing cells in the human body and are vital for metabolism, as well as intestinal and pancreatic function. They have been implicated in the pathogenesis of multiple diseases including diabetes mellitus. Although recent studies have identified multiple signaling pathways (including Wnt, MAPK, BMP and Notch) that can induce low levels of EE cell differentiation, the production of functional human EE cells in vitro remains challenging, making their study and therapeutic utilization difficult. To improve this, we employed the human intestinal organoid culturing system, as it mimics intestinal epithelial homeostasis, allowing for differentiation of multiple epithelial cell types. Using a small scale, directed screen, we targeted multiple transcriptional regulators, using small molecules known to control pancreatic and intestinal development, and hormone production. We chose small molecules instead of gene editing tools to avoid the potential pitfall of off-target mutagenesis.
We found that inhibition of FoxO1 in our organoid culture led to an increase in EE cell differentiation as assessed by EE-specific gene expression, with a 5-10 fold upregulation in expression of ChgA, NeuroD1, and Neurog3 compared to whole mucosal biopsies (P&lt;0.01 for all targets, n=3 per group). Flow cytometry data showed 6-8% of cells produced CHGA, compared to 0.2% in undifferentiated organoids (P&lt;0.0001, n=3 per group), and the 1% typically seen in the duodenum. We also noted a corresponding increase in the production of EE hormones, including glucose-dependent insulinotropic peptide (GIP), serotonin and somatostatin, by qPCR and immunofluorescence. Analysis of conditioned media using ELISA, compared to undifferentiated organoids, revealed increased serotonin (362.6±52.3 vs 167.5±5.1 ng/mL, P=.0037, n=3 per group) and GIP (5.76±1.31 pg/mL vs undetectable, n=3 per group). Independently, upregulation of GATA4-Nkx2.5 also induced EE cell differentiation and hormone production, although to a lesser extent than FoxO1 inhibition. The exception to this was GIP, which showed increased expression and production with GATA4-Nkx2.5 compared to FoxO1 inhibition (20.8±7.4 vs 5.8±1.3 pg/mL, n=3 per group), with a much larger increase when FoxO1 inhibition was followed by GATA4-Nkx2.5 activation (53.4±4.8 pg/mL, n=3). Of note, all experiments were performed in a minimum of three human lines.
Taken together, our data have identified multiple factors, including inhibition of FoxO1 and activation of GATA4-Nkx2.5, that can drive ex vivo human EE cell differentiation, with unique hormone production profiles, when targeted via small molecules. This is a critical first step towards understanding the role of enteroendocrine cells in disease and the development of EE cell-based therapies.
SAT-304 Pituitary Stem Cells May Drive Adenomas Causing Cushing’s Disease
Authors: Asuzu David , Johnson Kory , Elkahloun Abdel , Wu Weiwei , Nieman Lynnette , Stratakis Constantine A , Chittiboina Prashant 
Published: May 2020

Abstract
Introduction:Cell rests of self-renewing Sox2+ progenitor cells have been identified in the normal pituitary glands1, however their role in human pituitary tumorigenesis is not understood. Adrenocorticotropic hormone (ACTH) producing microadenomas that cause Cushing’s disease frequently (~70%) lack pathogenic genetic mutations.2 In mice, targeted expression of oncogenic β-catenin in Sox2+ cells generate microadenomas. Interestingly, the Sox2+ cells reside within the adjacent normal gland and drive adenomas in a paracrine fashion.3 We hypothesized that Sox2+ progenitors in human pituitary gland may drive the formation of microadenomas that cause Cushing’s disease (CD).
Methods:Four ACTH producing adenomas and two non-functional adenomas (NFPA) with separately annotated adjacent normal tissue (henceforward called ‘microenvironment’) were procured for this study (NCT00060541). We performed RNA deep sequencing (RNAseq) and compared expression of lineage-specific markers and progenitor markers using two-sample T-tests after testing for variance equality and using Welch’s approximation for degrees of freedom.
Results:We found expected overexpression of ACTH preprohormone POMC in CD adenomas compared to adjacent microenvironment (?-fold) and NFPA (?-fold). The microenvironment in Cushing’s disease showed increased expression of progenitor markers including SOX2, SOX9, CDH1, GRFA2, and KLF4 compared with microenviroment in NFPA. Likewise, the Cushing’s disease microenvironment showed increased expression ofPOMC (26.98 - fold, P = 0.004) as well as PRLR (FC 17.39, P = 0.006) and GH1 (FC 29.91, P = 0.003) implying that increased Sox2+ progenitors contribute to terminally differentiated corticotrope, lactotroph and somatotroph lineages in-vivo.
Conclusions:We report increased expression of several progenitor markers and concomitant elevation in tissues-specific markers in the microenvironment of Cushing’s disease patients. Our results indicate that increased pituitary progenitors in the microenvironment of human corticotropinomas may signal in paracrine fashion and may contribute to the pathogenesis of Cushing’s disease.
References:1. Cox, B. et al. J. Endocrinol.234, R135-R158 (2017).2. Bi, W. L. et al. Clin. Cancer Res.23, 1841-1851 (2017).3. Andoniadou, C. L. et al. Cell Stem Cell13, 433-445 (2013).
OR03-05 Novel Lipidome Signature in Active Cushing Syndrome Revealed by UHPLC-MS Metabolomics
Authors: Vega-Beyhart Arturo , Iruarrizaga Marta , Garcia-Eguren Guillermo , Pane Adriana , Giró Oriol , Aranda Gloria , Casals Gregori , Porta Mireia Mora , Halperin Irene , Carmona Francesc , Enseñat Joaquim , Vidal Oscar , Hu Ting , Rojo Gemma , Hanzu Felicia Alexandra 
Published: May 2020

Abstract
Background: Alterations in circulating amino acids, polyamines and acylcarnitines have been reported in patients with endogenous chronic hypercortisolism. However, lipid metabolites profiling and its interplay with the serum metabolome and degree of hypercortisolism in patients with active Cushing syndrome (CS) has not been previously assessed. Objective: To identify new metabolomic biomarkers associated with active CS.
Methodology: Multiple UHPLC-MS platforms were used to analyze the metabolome of serum samples obtained from 25 patients with active endogenous CS and 25 controls subjects matched by propensity score (sex, BMI, T2D, DLP, HBP).
Results: Metabolome of CS patients was deeply disrupted with 122 (27%) of the assessed metabolites significantly altered (p adj. &lt;0.05) out of which 5 bile acids resulted with the highest perturbation (&gt; 2-fold decrease). From the altered metabolites, 3 amino acids (AA), 2 acylcarnitines (ACs), 2 ceramides (CER) and 5 glycerophospholipids showed direction of effect independently associated with 24-h urinary free cortisol (MS) levels. A highly discriminant (AUC 96%) metabolome signature (n=59) characterized by lower levels of AA, ACs, polyunsaturated fatty acids (PUFA) and monoglycerophosphocolines (MGPC) together with increased levels of triacyclglycerols (TG), CER, diacylglycerophosphocholines (DGPC) and cholesteryl esters was identified and cross-validated (R2Y= 0.92, Q2Y= 0.68) using PLS-DA VIP scores &gt;1.5. PUFA omega-6, and alanine, aspartate and glutamate metabolism resulted the most impacted canonical pathways (q-stat 19.7, 10.8 (p&lt;0.001). Finally, topological network analysis detected 158 pairwise differential correlations (p &lt;0.005, 10,000-fold permutation) between 141 metabolites due to CS where the acylPC (P-18:1/0:0) resulted a key metabolite in the network (betweenness =0.117 & closeness centrality =0.467).
Conclusion: Active Cushing syndrome leads to a global proatherogenic shift in the circulating ceramides, glycerophospholipids and sphingolipids metabolites which are independently associated to the levels of urinary free cortisol being potential biomarkers of patients’ cardiovascular risk.
SUN-242 Transcriptome of the Amygdala in Normal Cyclic and Acyclic Gilts
Authors: Lents Clay A , Nonneman Dan J , Keel Brittney N 
Published: May 2020

Abstract
Many replacement gilts fail to express estrus, which results from being nonpubertal or behavioral anestrus. Genomic studies identified neuronal and olfaction pathways associated with these reproductive phenotypes. To better understand puberty failure, the transcriptome of the amygdala in nonpubertal, behavioral anestrus and normal cyclic gilts in the early follicular and midluteal phase was evaluated (n = 8/group; mean age = 259 d). An average of 57 million reads were obtained from each of 32 TruSeq mRNA libraries and mapped to Sscrofa 11.1. Differential expression of genes (DEG; adjusted P &lt; 0.05) was determined using DESeq2 and pathway analysis performed with iPathwayGuide. A total of 17,173 annotated genes were expressed (raw read count &gt; 15 in ≥ 8 samples). Comparing amygdala expression in nonpubertal gilts with follicular phase control gilts revealed 6 DEG (3 unannotated) including ARRDC2, ZFAND2A, and LAMC2. The only DEG identified between behavioral anestrus gilts and luteal phase control gilts was PIK3CG. There were 88 DEG in the amygdala of follicular phase gilts compared with luteal phase gilts. Expression of 73 genes was upregulated in the amygdala of follicular phase gilts and 14 genes were more highly expressed in luteal phase gilts. Enriched pathways included TGFβ signaling and ion transport related to GABAergic and glutamatergic neuronal function. Molecular processes for chemokine binding, membrane transporters and receptor signaling through phosphatidylinositol and tyrosine kinases were upregulated in the amygdala of follicular phase gilts. Prominent cellular components included ion channels and integrins necessary for focal adhesions that promote dendritic growth and neuronal synapse. Major differences in the amygdala of prepubertal and behavioral anestrus gilts were not found, with few genes differentially expressed compared to cyclic gilts. Stage of the ovarian cycle majorly impacted gene expression related to increased neuronal activity in the amygdala of follicular phase gilts. USDA is an equal opportunity provider and employer.
SUN-495 Rapid Resolution of Hyperthyroidism Induced Hepatic Dysfunction with Methimazole
Authors: Amir Maryam , Rahhal Marie-Noel , Calles-Escandon Jorge 
Published: May 2020

Abstract
Background:
Hepatic dysfunction in the setting of hyperthyroidism is difficult to diagnose and poses a challenge in therapy, since the classic medications used are potentially hepatotoxic.
Clinical case:
A 51-year-old female patient presented with fatigue, palpitations and tremors. BP 146/65 mmHg, HR 111 bpm and Temp 97.8 F. She was severely thyrotoxic with 3 + DTRs, tremors, enlarged thyroid gland with bruit on auscultation, clear lungs and no lower extremities edema. Blood tests showed TSH &lt;0.003 IU/ml (0.450-5.330 IU/ml), Free T4 5.33 ng/dL (0.45-1.80 ng/dL), Free T3 &gt; 30.0 pg/mL (2.3-4.2 pg/mL). Liver enzymes showed elevation in transaminases with ALT 319 IU/L (7-40 IU/L), AST 330 IU/L (7-40 IU/L) normal total Bilirubin 0.7 mg/dL (0.1-1.5 mg/dL) and Alkaline phosphatase 65 IU/L (40-200 IU/L). Transaminases were also elevated 3 weeks prior to presentation and this was extensively worked up with no identifiable etiology to explain the liver dysfunction. She was started on beta blocker therapy and admitted to the ICU. She had no clinical or echocardiographic evidence of cardiac dysfunction and remained hemodynamically stable. She was started on Methimazole 45 mg daily. The patient improved clinically and a pronounced decline in transaminase levels was documented in the first 72 hours. She was discharged home on day 3 of admission. On follow up visit her transaminases were found to have completely normalized within 14 days.
Discussion:
The diagnosis of elevated transaminases in hyperthyroidism is a challenge. This is due to the possibility of multiple etiologies including decreased cardiac output and/or liver congestion, concomitant primary liver disease or more specifically autoimmune hepatic disease such as primary biliary cirrhosis. (1-2). Incidentally Methimazole has been associated with transient asymptomatic transaminitis typically during the first three months of therapy (3). Our case indicates, that methimazole can be used in patients with a presumed diagnosis of hyperthyroidism induced hepatotoxicity after all possible etiologies are ruled out. Hyperthyroidism as the cause of liver dysfunction can only be entertained after all those etiologies are ruled out and upon resolution of transaminase elevation in conjunction with improvement in the hyperthyroid state itself, as was demonstrated in this case.
Conclusion:
Methimazole can be safely used in patients with severe hyperthyroidism and elevated liver enzymes when all other etiologies of liver dysfunction have been ruled out.
References:
1. Khemichian S, Fong TL. Hepatic dysfunction in hyperthyroidism. Gastroenterol Hepatol (N Y).
2. Elias RM, Dean DS, Barsness GW. Hepatic dysfunction in hospitalized patients with acute thyrotoxicosis: a decade of experience.
3. LiverTox: Clinical and Research Information on Drug-Induced Liver Injury. Bethesda (MD): Methimazole
SAT-300 Evading Death: Noxa in Cushing’s Disease Pituitary Adenomas
Authors: Alvarez Reinier , Floros Konstantinos , Johnson Kory , Elkahloun Abdel , Wu Weiwei , Edwards Nancy , Chaudhury Abhik Ray , Nieman Lynnette , Stratakis Constantine , Chittiboina Prashant 
Published: May 2020

Abstract
Introduction: Recurrence of Cushing’s disease (CD) caused by benign pituitary microadenomas are challenging clinical problems. Mechanisms underlying adenoma formation and recurrence remain unknown. PMAIP1 gene codes for Noxa, a Bcl-2 homology 3 (BH3) pro-apoptotic protein frequently downregulated in malignant human tumors.1-6 The role of dysregulated apoptosis remains largely unknown in benign tumors and in CD. We hypothesized that altered expression of Noxa protein is a pro-survival adaptation employed by CD adenomas.
Methods: Syngeneic human pituitary adenoma and adjacent normal gland pairs (n=2), and an additional CD adenoma were analyzed with RNAseq. 10 CD, 1 growth hormone (GH) and 1 non-functioning adenoma (NFPA) underwent immunohistochemical (IHC) analysis for Noxa expression, which was graded by a neuropathologist as 0=none, 1=light, 2=medium, 3=strong. Staining grade represents relative protein expression.
Results: Compared to adjacent normal pituitary tissue, we found that adenomas (n = 3) had a 3.76 fold increase in PMAIP1 mRNA. However, there was attenuated Noxa IHC staining in adenomas compared to normal pituitary in 8 of 10 CD patients (2:3, respectively), but similar staining in 2 of 10 CD patients (2:2 and 2-3:2-3). In GH and NFPA, we found similar patterns of Noxa suppression in the adenomas compared to the normal gland.
Conclusion: Despite elevated PMAIP1 (Noxa) gene expression in adenomas compared to adjacent normal gland in CD, protein expression was reduced in adenomas. This downregulation of Noxa protein expression may contribute to reduced apoptosis of tumor cells. These findings suggest that CD adenomas gain pro-survival advantage by downregulating Noxa protein at post-transcriptional or post-translational level.
References 1. Escobar, D. et al. Cell Death Dis.6, 1-14 (2015).2. Brinkmann, K. et al. Cell Rep.3, 881-891 (2013).3. Liu, Y. L. et al. Oncotarget5, 11237-11251 (2014).4. Dengler, M. A. et al. Cell Death Dis.5, 1-10 (2014).5. Liang, L. et al. J. Oral Pathol. Med.48, 52-59 (2019).6. Tahir, S. K. et al. Cancer Res.67, 1176-1183 (2007).
SAT-022 Adrenal Androgen Production Is Maintained While Ovarian Estrogens Fall Following the Final Menstrual Period in the Study of Women’s Health Across the Nation (SWAN)
Authors: McConnell Daniel S , Lasley Bill L , Crawford Sybil , Gee Nancy A , Auchus Richard Joseph 
Published: May 2020

Abstract
The aim of this study was to clarify changes in sex steroids at the final menstrual period (FMP). We have shown previously that estradiol (E2) declines substantially in the 4-year period around the FMP, but hypothesize that testosterone (T) declines modestly and adrenal Δ5 androgens dehydroepiandrosterone (DHEA) and androstenediol (Adiol) remain unchanged. Methods: Liquid chromatography tandem mass spectrometry (LC-MS/MS) and immunoassay was used in approximately annual samples collected before and following FMP in 1490 women. We estimated time-related changes in each log-transformed androgen using piecewise linear mixed modeling, with knots (slope changes) at FMP-2 yrs and FMP+2 yrs as seen for E2. These models then were re-estimated for subgroups with different time courses identified using group-based trajectory modeling. Results: In the full sample, T was generally stable, although time course varied by subgroup, with a significant decrease of 5%/year in T in [FMP-2yrs, FMP+2yrs] only in the lowest T women. For DHEA and Adiol, declines were similar across all 3 time segments and across subgroups. Mean circulating androgen concentration declined modestly (P&gt; 0.05) from five years before to five years following FMP. However, when stratified only the lowest 7% of circulating T declined significantly (p&lt; 0.05) in the four years surrounding FMP when mean circulating E2 declined. This trajectory divergence of the lower circulating T suggests a different, non-adrenal source that is decreased at FMP which may be useful in clarifying ovarian versus adrenal testosterone production during the post-menopause. Paired results from samples collected before and following FMP in the same subjects indicate mean circulating E2 is less than 5% of mean circulating T suggesting that a relatively large portion of circulating E2 may be largely a result of peripheral conversion of adrenal androgens. Longitudinal LC-MS/MS analyses of circulating E2 and T indicate that the principal change in sex steroid influence at menopause is largely a decrease and dampening of ovarian and not adrenal steroid production.
SUN-313 Depression, Subjective Stress and Serum Osteocalcin Concentrations in People with Type 2 Diabetes Mellitus
Authors: Nguyen Michelle My Linh , Darwish Lina , Major-Orfao Chelsi , Colby-Milley Jessica , Wong Sophie K , Sugamori Kim , Lanctôt Krista L , Herrmann Nathan , Oh Paul I , Yang Pearl , Shah Baiju R , Gilbert Jeremy , Assal Angela , Halperin Ilana Jaye , Mitchell Jane , Swardfager Walter 
Published: May 2020

Abstract
Serum osteocalcin is emerging as a potential marker of glucose homeostasis and risk of type 2 diabetes mellitus (T2DM). In mice, osteocalcin knockouts exhibit depressive-like behaviours but knockouts of a putative osteocalcin receptor, GPR158, are resistant to chronic unpredictable mild stress. Recently, osteocalcin release from bone has been suggested to mediate some aspects of the acute stress response. Here, we assess relationships between serum osteocalcin, depression and perceived stress in people with T2DM. Participants with T2DM were assessed for whether they met the DSM-5 criteria for Major Depressive Disorder using the research version of the Structured Clinical Interview for DSM-5 depression criteria (SCID-5RV). Subjective stress was assessed using the Perceived Stress Scale (PSS) with higher scores indicating greater subjective stress. Serum carboxylated (cOCN) and uncarboxylated (unOCN) osteocalcin were assayed from fasting morning blood by commercial ELISA. Among 87 participants (mean age 62.9±9.5, 52% women), 18 (26%) were experiencing a depressive episode (7 men, 11 women). Both serum unOCN and cOCN were associated with higher PSS scores in participants with depression (unOCN, r=0.566, p=0.014; cOCN, r=0.564, p=0.015) but not in those without depression (unOCN, r=0.002, p=0.985; cOCN, r=0.090, p=0.463). A significant interaction was found between depression and PSS scores predicting serum unOCN in a linear model adjusted for age, sex, body mass index, antidepressant use and HbA1c (F=6.225, p=0.015). The results are consistent with reports that osteocalcin release from bone may be a mediator of stress perception; however, among people with T2DM, this relationship was observed only among those currently experiencing a depressive episode.
SUN-278 Isolated Hypothalamic Langerhans Cell Histiocytosis in an Adult Manifesting as Panhypopituitarism
Authors: Mikhael Alexandra , Gautam Nitesh , Buehler Lauren Anne , Skugor Mario 
Published: May 2020

Abstract
Background: Langerhans Cell Histiocytosis (LCH) is a rare disease characterized by abnormal proliferation of bone marrow derived histiocytes. Although predominantly a childhood disease, LCH can occur at any age and has an incidence of one to two cases per million in adults. LCH can involve a single organ or present as disseminated disease. Sites most commonly affected are bone, skin, bone marrow and pituitary. Isolated hypothalamic LCH is a rare entity. A few reports of LCH presenting as a hypothalamic mass exist but mainly in children. Treatment of adult LCH is derived from pediatric studies, and no standard of care exists. We report a unique case of an adult with panhypopituitarism secondary to a hypothalamic mass found to be biopsy proven isolated LCH treated with Vemurafenib, a BRAF V600E inhibitor.
Clinical Case: A 66-year-old previously healthy man presented with progressively increasing confusion, cognitive decline and generalized weakness requiring hospital admission. Laboratory evaluation revealed an 8AM cortisol of 2.5 ug/dL (reference [ref] 7.0–25.0 ug/dL), ACTH of 9.3 pg/mL (ref 8–42 pg/mL), TSH of 0.106 uIU/mL (ref 0.35–5.5 uIU/mL), free T4 of 0.67 ug/dL (ref 0.7–1.25 ng/dL), LH &lt;0.2 IU/L (ref 0.5–11 IU/L), FSH 0.6 IU/mL (ref 1–12 IU/L), total testosterone &lt;12 ng/dL (ref 193 - 824 ng/dL), free testosterone &lt;3.3 pg mL (ref 41.7 - 180.2 pg/mL), IGF1 41 ng/mL (ref 33–220 ng/mL) and prolactin of 31.5 ng/mL (ref 3.5–15 ng/mL). Findings were consistent with panhypopituitarism, and he was started on glucocorticoid and levothyroxine replacement therapy. MRI pituitary revealed an 18.1 x 13.8 x 15.8 mm hypothalamic mass with signal intensity alteration in the optic pathway. Patient developed polyuria with a serum sodium of 148 mmol/L (ref 136–145 mmol/L), elevated serum osmolality and low urine osmolality consistent with central DI and was started on oral DDAVP.
The patient underwent right sided neuroendoscopic intraventricular biopsy of hypothalamic mass. Pathology was consistent with LCH with immunohistochemistry staining for CD1a, Langerin and BRAFV600E, with occasional cells staining for CD68, CD163 and S100. Bone marrow biopsy was normal. A NM PET scan revealed a hypermetabolic hypothalamic mass compatible with LCH without evidence of involvement of other sites including the skeletal system. Given histopathologically confirmed BRAF-mutated LCH, treatment with Vemurafenib was initiated. Behavioral changes gradually improved, and repeat MRI brain three months after treatment revealed decrease in size of hypothalamic mass.
Conclusion: Isolated hypothalamic LCH is exceedingly rare. No clear guidelines exist for treatment of LCH in adults. BRAFV600E mutations are found in more than fifty percent of LCH cases and are associated with worse outcomes. This case demonstrates the successful use of targeted BRAF inhibitor therapy in an adult patient with BRAF mutated hypothalamic LCH.