bluuebunny/crossref_metadata_2025 huggingface dataset:
174-LB: Association between Metformin Treatment and Improved Symptoms of Post-Traumatic Stress Disorder
Authors: LIU SHUQIAN , RAINES AMANDA M. , YOSHIDA YILIN , TANG WAN , CONSTANS JOSEPH I. , FONSECA VIVIAN , SHI LIZHENG
Published: June 2020

U.S. veterans have a high prevalence of post-traumatic stress disorder (PTSD), for which no effective psychotropic medications are indicated. Nearly 25% of veterans have diabetes, for which metformin is a first line treatment. Although metformin is associated with improved neurodegenerative outcomes, there is no evidence on the association between metformin and PTSD-related symptom relief. This study aimed to examine whether metformin was associated with a clinically meaningful reduction in PTSD symptoms (≥20 points reduction PTSD checklist for DSM-5 {PCL-5}) score)) among veterans with PTSD using data from Veterans Health Administration. The metformin cohort was defined as first metformin use after a recorded PTSD diagnosis with a PCL-5 score ≥33 (i.e., clinical cut-score). The control cohort (never used metformin) was selected by propensity score 1:1 matching on the likelihood of initiating metformin with the covariates of age, gender, race, durations of PTSD, diabetes, and depression, and baseline CCI, PCL-5, and PHQ-9 scores. Multivariable Cox proportional regression was used to assess the association between metformin exposure and improvement in the PCL-5 scores. A total of 7950 veterans (mean age: 51.9 ± 13.6 years, male: 85%, white: 61.4%, African American: 28.8%) with confirmed PTSD were comparable on the baseline demographics and medical conditions between metformin and control PSM-matched cohorts with a median follow-up 1.2 years in both cohorts. The baseline PCL-5 score was 56.2±11.6 and 56.5±11.7 in metformin and control cohort, respectively. The metformin-treated cohort was more likely to have a clinically meaningful reduction in PTSD symptoms (adjusted hazard ratio (aHR): 1.31, 95%CI: 1.16-1.48), compared the control cohort. Furthermore, patients on metformin ≥2 years were 22% more likely to reduce PTSD symptoms (aHR: 1.22, 95%CI: 1.04-1.43), compared with metformin exposure <2 years. Metformin exposure may be beneficial to PTSD symptoms among veterans.

Disclosure

S. Liu: None. A.M. Raines: None. Y. Yoshida: None. W. Tang: None. J.I. Constans: None. V. Fonseca: Consultant; Self; Abbott, Asahi Kasei Corporation, AstraZeneca, Bayer Inc., Novo Nordisk Inc., Sanofi. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc. Stock/Shareholder; Self; Amgen, Bravo4health. L. Shi: None.
1742-P: Targeting the Low Molecular Weight Protein Tyrosine Phosphatase for Obesity-Associated Diabetes Therapy
Authors: STANFORD STEPHANIE M. , DIAZ MICHAEL A. , ZOU JIWEN J. , ARDECKY ROBERT J. , PINKERTON ANTHONY , BOTTINI NUNZIO
Published: June 2020

Obesity-associated insulin resistance plays a central role in the pathogenesis of type 2 diabetes. A promising approach to decrease insulin resistance in obesity is to inhibit the protein tyrosine phosphatases that negatively regulate insulin receptor signaling. The low molecular weight protein tyrosine phosphatase (LMPTP), encoded by the ACP1 gene, acts as a critical regulator of insulin signaling by dephosphorylating the activation motif of the insulin receptor. Human genetics studies suggest that high LMPTP catalytic activity promotes diabetes and insulin resistance in obesity. We recently discovered that LMPTP is a critical promoter of insulin resistance in obesity through an action on the liver and that pharmacological inhibition of LMPTP reverses obesity-induced diabetes in mice. Here we report our progress in the development of a new chemical series of LMPTP inhibitors that is orally bioavailable and effective at treating disease in obese diabetic mice.

Disclosure

S.M. Stanford: None. M.A. Diaz: None. J.J. Zou: None. R.J. Ardecky: None. A. Pinkerton: None. N. Bottini: Research Support; Self; Eli Lilly and Company, Gilead Sciences, Inc., Kyowa Hakko Kirin Co., Ltd., Roche Pharma.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-15-INI-13 to S.M.S.); National Institutes of Health (R01DK106233)
1744-P: Hepatic and Disposal Insulin Sensitivity with Single Tracer Method: Use of Naturally Abundant 13C-Polysaccharide
Authors: MAN CHIARA DALLA , VISENTIN ROBERTO , SCHIAVON MICHELE , BASU ANANDA , COBELLI CLAUDIO , BASU RITA
Published: June 2020

We have recently demonstrated successful estimation of postprandial glucose fluxes using a novel triple tracer technique after ingestion of a mixed meal containing complex carbohydrates (CC) that is naturally enriched with 13C polysaccharide ([13C]-PS). We also reported higher insulin sensitivity (SI) with CC than simple carbohydrate (SC) containing mixed meals. The current analysis was designed to determine if [13C]-PS per se could directly measure hepatic (SIH) and disposal SI (SID) during the CC containing mixed meal. Sixteen healthy subjects (9M, age=33±2 y, BMI=23±1 kg/m2) received, in random order, either SC (labeled with [13C]-glucose) or CC (naturally containing [13C]-PS) mixed meals. All meals were isocaloric and contained 50g carbohydrates with similar macronutrient composition. Plasma glucose, [13C]-glucose and insulin were measured and the Single Tracer Oral Minimal Model applied to estimate SIH and SID. The model well predicted the data. SIH was significantly higher, while SID was numerically but not statistically higher in CC than SC meals (Figure 1). This implies that the higher SI with CC meals is likely due to higher hepatic insulin sensitivity. Taken together, the data suggest that a single, naturally occurring [13C]-PS tracer can measure postprandial SI and its components without the need for the triple tracer method.

Disclosure

C. Dalla Man: Research Support; Self; Sanofi-Aventis Deutschland GmbH. R. Visentin: None. M. Schiavon: None. A. Basu: Consultant; Spouse/Partner; GENFIT. Research Support; Spouse/Partner; AstraZeneca. C. Cobelli: None. R. Basu: Consultant; Self; GENFIT. Research Support; Self; AstraZeneca.

Funding

National Institutes of Health (DK085516, DK94331, DK29953); Italian Minister for Education
1749-P: The TNFa Inhibitor Infliximab Decreases Inflammation in High-Fat Fed Mice but Does Not Alleviate Insulin Resistance
Authors: ROBY DEBORAH A. , GUAN CHRISTOPHER , JAMES FREYJA , BRACY DEANNA , LANTIER LOUISE , WASSERMAN DAVID
Published: June 2020

The insulin resistance of diet-induced obesity (DIO) is accompanied by inflammation. Previous studies suggest that the anti-inflammatory effects of infliximab, an anti-TNFα chimeric monoclonal antibody, decreases DIO-associated inflammation in mice fed a 60% fat diet. The effectiveness of infliximab at 10 mg/kg Infliximab (IF10) and 100 mg/kg infliximab (IF100) in C57Bl/6J DIO mice was studied in comparison to C57Bl/6J DIO mice administered chimeric IgG (IgG; 10 mg/kg). Treatment consisted of IP injections of infliximab or IgG once per week for 4 weeks. Body weight and fasting arterial glucose was not different in IF10, IF100, and IgG. IF10 and IF100 mice had a decreased presence of crown-like structures in white adipose tissue compared to IgG treated mice. Infliximab resulted in a dose-dependent reduction in macrophage F4/80 staining in white adipose tissue (4.3±0.7% and 1.8±0.2% in IF10 and IF100, respectively) compared to 8.1±1.6% in IgG, validating the anti-inflammatory effect. Insulin action was measured in mice with arterial and venous catheters using the hyperinsulinemic, euglycemic clamp method (insulin dose of 4 mU/kg/min) combined with [3-3H]glucose in conscious, unstressed mice. Glucose infusion rates (GIR) in IgG mice were higher (25±3 mg/kg/min) as compared to IF10 (17±2 mg/kg/min) and IF100 (19±2 mg/kg/min). The greater GIR in IgG as compared to IF10 was due to changes in hepatic endogenous glucose production (suppression of 12±3 mg/kg/min and 5±1 mg/kg/min, respectively) without a difference in glucose utilization. IF100 exhibited small reductions in both hepatic and peripheral insulin action, neither of which reached significance. Mice treated with saline alone were used as an additional control, but neither IF10 nor IF100 had significant effects on insulin action compared to saline controls.

In summary, despite profound anti-inflammatory effects of the anti-TNFα drug, Infliximab causes a paradoxically greater hepatic insulin resistance.

Disclosure

D.A. Roby: None. C. Guan: None. F. James: None. D. Bracy: None. L. Lantier: None. D. Wasserman: None.
176-LB: Ethnic Differences in Cardiovascular Complications of Young-Onset Diabetes (YOD)
Authors: KE CALVIN , STUKEL THERESE , LUK ANDREA , CHAN JULIANA C. , SHAH BAIJU R.
Published: June 2020

Introduction: YOD (age at diagnosis [AAD]<40 years) is associated with greater cardiovascular disease (CVD) risk than usual-onset diabetes (UOD, AAD≥40 years). It is unknown whether this pattern varies by ethnicity.

Methods: We included adults in Ontario, Canada with incident diabetes (2002-12, followed to 2018). We matched each case with ≤5 diabetes-free controls by age, sex, and ethnicity. The outcome was CVD (coronary artery disease, congestive heart failure, stroke, peripheral revascularization, lower extremity amputation). We constructed Cox proportional hazards models to estimate the association between AAD and CVD relative to controls.

Results: We included 411,030 cases (3.5% South Asian; 4.3% Chinese) and 2,000,035 controls. The hazard ratios (HR) for CVD in UOD versus controls were higher in Chinese (2.38, 2.2-2.5) and South Asian (2.5, 2.4-2.7) people than White people (1.9, 1.9-1.9; pinteraction<0.0001). The HR for CVD in YOD versus controls were similar across ethnicities (Chinese: 5.2, 3.6-7.3; South Asian: 4.4, 3.7-5.3; White: 4.6, 4.4-4.8). In YOD and controls, Chinese and South Asian people had lower hazard of CVD than White people. In UOD, CVD hazard was lowest in Chinese people, and similar in South Asian and White people.

Discussion: Although Chinese and South Asian people have a lower hazard of CVD compared to White people, YOD is associated with a 4- to 5-fold greater hazard of CVD across ethnicities.

Disclosure

C. Ke: None. T. Stukel: None. A. Luk: Research Support; Self; Bayer Healthcare Pharmaceuticals Inc., Roche Pharma. Other Relationship; Self; Merck Sharp & Dohme Corp. J.C. Chan: None. B.R. Shah: None.

Funding

Canadian Institutes of Health Research; South Asian Network Supporting Awareness and Research
1761-P: MC4R-Expressing Neurons in the PVH Regulate Glycemia Independent of Insulin
Authors: UNER AYKUT G. , LEE HYON , HAN JUNG SUK , PEREIRA RODRIGO M. , SUN XIAOFANG , KIM HYUN JEONG , KIM YOUNG-BUM
Published: June 2020

Hypothalamic and extra-hypothalamic neuronal populations play important roles in the regulation of energy balance and glucose homeostasis. However, the underlying mechanisms of this control remain unclear. We have recently reported that inhibition of hypothalamic pro-opiomelanocortin (POMC) neuronal activity leads to a reduction in blood glucose levels of normoglycemic POMC-cre mice. This effect is independent of energy intake. Since hypothalamic POMC neurons project to the paraventricular nucleus of the hypothalamus (PVH) and regulate energy metabolism via melanocortin-4 receptor (MC4R), we hypothesized that changes in the activity of MC4R-expressing neurons in the PVH could alter glycemic levels. To address this, we stereotaxically injected designer receptors exclusively activated by designer drug (DREADD) viruses into the PVH of MC4R-cre mice to chemogenetically and selectively activate or inhibit these neurons. We continuously activated or inhibited these neurons by injecting clozapine-N-oxide (CNO) every 8 hours for 5 days and monitored daily blood glucose levels and energy intake. Interestingly, inhibition of MC4R-expressing neurons in the PVH leads to a significant reduction (∼ 30%) in blood glucose levels despite of increased food intake. On the other hand, blood glucose levels are significantly increased, but food intake is greatly decreased when MC4R-expressing neurons are activated. We further determined the peripheral insulin response via MC4R-expressing neurons in the PVH. We found that CNO-induced inhibition or activation of MC4R-expressing neurons did not affect serum insulin levels. These data suggest that MC4R-expressing neurons in the PVH can regulate blood glucose levels and food intake via divergent cellular mechanisms, which is independent of circulating insulin. Thus, the inhibition of MC4R-expressing neurons may be a promising target for novel antidiabetic medicines.

Disclosure

A.G. Uner: None. H. Lee: None. J. Han: None. R.M. Pereira: None. X. Sun: None. H. Kim: None. Y. Kim: None.
1762-P: Poor Sleep Quality Is Associated with Lower Absolute Cerebral Glucose Levels
Authors: STANLEY THEODORA K. , GUNAWAN FELONA , REDEKER NANCY S. , JIANG LIHONG , COPPOLI ANASTASIA , ROTHMAN DOUGLAS L. , MASON GRAEME F. , HWANG JANICE
Published: June 2020

Poor sleep quality has been associated with increased risk of metabolic syndrome and type 2 diabetes as well as acceleration of neurodegenerative diseases. In rodents, sleep restriction decreases glucose transport into the brain via downregulation of GLUT1, the primary glucose transporter at the blood-brain barrier. However, little is known about the association between sleep quality and glucose transport and metabolism in the human brain. In this exploratory analysis, we quantified cerebral glucose levels amongst individuals with good and poor sleep quality as defined by the widely used and well-validated Pittsburgh Sleep Quality Index (PSQI), which assesses 7 different components of sleep quality over a 1 month time period (PSQI≥5 = Poor sleep; PSQI<5 = Good sleep). Twelve healthy subjects completed the PSQI questionnaire. Eight (6F, age 27.6 ± 5.7, BMI 26.6 ± 8.2 kg/m2, HgbA1c 5.4 ± 0.2%) had good and 4 (2F, age 29.5± 2.4, BMI 27.8 ± 6.5 kg/m2, HgbA1c 5.2 ± 0.2%) had poor sleep. All subjects underwent 13C magnetic resonance spectroscopy brain scanning at 4 Tesla during a 2-hour hyperglycemic clamp (plasma glucose target ∼180 mg/dl) to measure absolute cerebral glucose levels as part of a larger study to investigate obesity and cerebral glucose metabolism. There were no differences in age, BMI, or HbA1c levels between groups with good and poor sleep. Individuals with good sleep had 63% higher absolute cerebral glucose levels at steady state compared to those with poor sleep (3.4 ± 0.7 mmol/L vs. 2.1 ± 0.9 mmol/L, p=.017). Higher PSQI scores correlated with lower absolute cerebral glucose levels (r= -0.725, p=0.008, PSQI for all subjects (mean±SD) = 4 ± 2). In this pilot and exploratory study, individuals with poor sleep quality have significantly lower absolute cerebral glucose levels, which suggests an association between poor sleep and altered cerebral glucose transport and/or metabolism. These findings may have wide-ranging implications for understanding the effects of sleep on brain function.

Disclosure

T.K. Stanley: None. F. Gunawan: None. N.S. Redeker: None. L. Jiang: None. A. Coppoli: None. D.L. Rothman: None. G.F. Mason: None. J. Hwang: Research Support; Self; General Electric.

Funding

American Diabetes Association (1-17-ICTS-013 to J.H.); National Institutes of Health (1R03DK121048)
1774-P: Neurally Mediated Prandial Islet-Cell Function Is Glucose-Independent and Preserved after Gastric Bypass and Sleeve Gastrectomy
Authors: HONKA HENRI , AL ZUBAIDI ANAS M. , DEFRONZO RALPH A. , GASTALDELLI AMALIA , SALEHI MARZIEH
Published: June 2020

The weight-loss independent glycemic effects of gastric bypass (GB) are characterized by prandial short-lived hyperinsulinemia as a result of β-cell stimulation by glucose and incretins. To investigate the effect of parasympathetic nervous system (PNS) activity, a neural component of the enteroinsular axis, on islet function, 9 subjects with history of GB, 6 with prior sleeve gastrectomy (SG), and 4 non-surgical controls were recruited. The GB, SG, and CN groups were matched for BMI, FFM, A1C, and age; and the GB and SG for weight loss and time postsurgery; none had diabetes. To measure islet hormone responses to meal ingestion when the effect of GI hormones and glycemia is minimal, subjects consumed a liquid meal (280 kcal, 20% glucose) at 2 hr of a 5-hr hyperinsulinemic (120 mU/m2/min) hypoglycemic (55mg/dl) clamp with and without atropine infusion on two separate studies. Fasting levels of glucose, islet and gut hormones were similar among 3 groups and 2 studies. Despite similar insulin sensitivity among the groups the insulin clearance estimated before meal ingestion was larger in surgical patients (P=0.08). Suppression of endogenous β-cell secretion by hypoglycemia was less in both surgical groups compared to controls (P<0.001), but it was reduced by atropine in 3 groups (P<0.05). Hypoglycemia-induced glucagon secretion was not affected by PNS blockade. Blocking PNS diminished both β- and α-cell secretory response to meal ingestion (P < 0.05) in both surgical and non-surgical groups. Pre- and post-meal pancreatic polypeptide, marker of vagal input to the islet, declined to baseline values during atropine infusion. Meal-induced GIP secretion was similar in studies with and without atropine whereas blocking PNS significantly reduced GLP-1 secretion (P < 0.05) in all subjects. Our data shows that in nondiabetic subjects with prior GB and SG, PNS activity regulates islet and gut hormone secretory response to meal ingestion independent of glucose levels.

Disclosure

H. Honka: None. A.M. Al Zubaidi: None. R.A. DeFronzo: None. A. Gastaldelli: Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, Gilead Sciences, Inc., Inventiva Pharma, Novo Nordisk Inc. M. Salehi: None.

Funding

Finnish Cultural Foundation (00180071); National Institutes of Health (DK105379)
1775-P: Molecular Dissection of Brainstem GABA Neurons Regulating Energy Balance
Authors: BHAVE VARUN M. , KAMATKAR NACHIKET , NECTOW ALEXANDER R.
Published: June 2020

An important goal in metabolism is to identify brain regions capable of regulating energy balance. These brain regions represent potential druggable targets for the modification of food intake and energy expenditure, which could help to combat any number of debilitating disorders relating to metabolic function and body weight. We have recently identified the dorsal raphe nucleus (DRN) as a key node in the central nervous system regulating both food intake and energy expenditure. In particular, we have found that inhibitory, GABAergic neurons residing in the dorsolateral aspect of this nucleus are activated by hunger and in turn induce the appropriate counterregulatory responses – namely, to increase food intake and suppress energy expenditure. Here, using a combination of ensemble-level molecular profiling in tandem with single-nuclear RNA sequencing, we demonstrate that these GABAergic neurons are molecularly heterogeneous and express a host of factors directly implicated in the central control of energy homeostasis. Of note, a few of these subpopulations express abundant levels of transcript for neuropeptides important in regulating energy balance, such as thyrotropin-releasing hormone (Trh) and neuropeptide Y (Npy). These profiling studies thus demonstrate a previously underappreciated level of heterogeneity within GABAergic neurons of the DRN suggesting a multifaceted, and possibly distributed, mechanism through which these neurons may regulate energy balance. Together, this work molecularly characterizes the DRN at a resolution not previously possible and identifies cell subpopulations through which this nucleus may regulate energy homeostasis.

Disclosure

V.M. Bhave: None. N. Kamatkar: None. A.R. Nectow: None.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-18-ACE-49 to A.R.N.); American Diabetes Association (1-18-IBS-148 to A.R.N.); Foundation for Prader-Willi Research; National Alliance for Research on Schizophrenia & Depression
1777-P: Defining VDR Expression in the Brain Using a Novel VDR-Cre Mouse Line
Authors: LIU HAILAN
Published: June 2020

The central nervous system mediates the effects of vitamin D on energy and glucose balance, but the distribution of vitamin D receptor (VDR) in the brain is not clear due to the lack of reliable antibodies. Here, we generated a knock-in VDR-Cre mouse line, in which cre recombinase activity is driven by the endogenous VDR promoter. By mating VDR-Cre mice with TdTomato reporter mice, we found that VDR-expressing neurons are located in multiple brain regions, including the cortex, hypothalamus, amygdala and caudate putamen. We further verified the specificity of our model through single cell RT-PCR, which showed significantly higher vdr expression in tomato+ cells. More importantly, our in vitro electrophysiological study revealed that vitamin D can selectively activate tomato+ neurons, without affecting tomato- neurons. By using immunofluorescence staining, we found a small portion of VDR positive neurons also express oxytocin, ß-endorphin, ERα or serotonin. Interestingly, in the ventromedial hypothalamus, ERα co-localized with only 1% tomato+ neurons in males but 8% tomato+ neurons in females. Taken together, we created and validated a novel VDR-Cre mouse line, which provides a useful tool for future studies to investigate the functions of VDR in the brain.

Disclosure

H. Liu: None.
1778-P: A Single Cell Map of Metabolically Regulated Neurons in the Ventromedial Hypothalamic Nucleus
Authors: RUPP ALAN , AFFINATI ALISON , MYERS MARTIN G.
Published: June 2020

While the ventromedial hypothalamic nucleus (VMN), especially the dorsomedial VMN (dmVMN), plays important roles in the control of blood glucose and energy balance, the subpopulations of VMN neurons that comprise the dmVMN have remained poorly defined. To identify dmVMN subpopulations in an unbiased manner, we performed single nucleus RNA-seq on mouse VMN and clustered the VMN neurons into subtypes based upon their transcriptional profiles (T-types). This analysis revealed over a dozen distinct T-types that can be organized hierarchically into 5 major classes of neurons that exhibit distinct anatomic distributions within the VMN (dm, ventrolateral, and tuberal). Several dmVMN T-types expressed marker genes known to contribute to the control of energy balance and glucose homeostasis (e.g., Lepr, Tcf7l2), and many of these genes were expressed in distinct T-types, suggesting the existence of multiple T-types that modulate metabolism, each of which may play distinct roles in metabolic control. Indeed, exposure to high-fat diet induced obesity (DIO) provoked transcriptional changes only in a subset of VMN T-types. Furthermore, while some DIO-induced transcriptional changes were common to all affected T-types, many changes were unique to a single T-type. Together, these findings not only identify the distinct subpopulations of dmVMN neurons in an unbiased manner, but they also reveal specific dmVMN subpopulations that respond to the metabolic perturbations associated with obesity, as well as the unique transcriptional responses of each subpopulation to these perturbations.

Disclosure

A. Rupp: Research Support; Self; Novo Nordisk A/S. A. Affinati: Research Support; Self; Novo Nordisk Inc. M.G. Myers: Research Support; Self; AstraZeneca, Novo Nordisk Inc.
1784-P: Liraglutide Treatment Stimulates Alpha Cells to Produce Glucagon-Like Peptide-1 via Expression of Prohormone Convertase 1/3 in Mice and Human
Authors: SAIKIA MRIDUSMITA , HOLTER MARLENA M. , GARIBAY DARLINE , GARCIA-OCANA ADOLFO , DANKO CHARLES G. , CUMMINGS BETHANY
Published: June 2020

Glucagon-like peptide-1 (GLP-1) regulates islet function by enhancing glucose-stimulated insulin secretion (GSIS). We previously found that the beta cell GLP-1 receptor (GLP-1R) contributes to improvements in glucose regulation and islet function in mice that have undergone bariatric surgery, specifically, vertical sleeve gastrectomy (VSG). Further investigation revealed that VSG increases alpha cell GLP-1 production by turning on the expression of prohormone convertase 1/3 (PC1/3, gene: Pcsk1) in a beta cell GLP-1R-dependent fashion. These findings lead us to hypothesize that beta cell GLP-1R signaling increases alpha cell GLP-1 production to augment GSIS in a paracrine positive feedback loop. In the current study, we tested whether the effects of VSG can be recapitulated by pharmacologic stimulation of the GLP-1R, using a GLP-1 analog, liraglutide. We found that liraglutide treatment increases alpha cell GLP-1 expression in a beta cell GLP-1R dependent fashion (GLP-1 area per islet (µm2): Vehicle WT = 132 ± 3, Vehicle KO = 73 ± 2, Liraglutide WT = 241 ± 15, Liraglutide KO = 67 ± 1; P<0.05 Liraglutide WT vs. all groups). We then used an innovative single-cell RNA-sequencing platform, DART-seq, to determine the translational relevance of this new model of islet function. We validated application of DART-seq in human islets, and using this technology we observed that liraglutide induces a 1.5 log-fold increase in Pcsk1 mRNA expression compared with saline treated controls in a subset of alpha cells (P<0.05 saline vs. liraglutide). Additionally, our DART-seq data suggests that liraglutide-induced increase in alpha cell PC1/3 expression may be part of a broader induction of alpha-cell transdifferentiation to beta cells. Thus, our study demonstrates that the effect of beta cell GLP-1R signaling to increase alpha-cell GLP-1 expression can be stimulated by pharmacological activation of the GLP-1R and has translational relevance in humans.

Disclosure

M. Saikia: None. M.M. Holter: None. D. Garibay: None. A. Garcia-Ocana: None. C.G. Danko: None. B. Cummings: None.
1807-P: MR-Based Assessments Better Capture Pathophysiologic Profiles in Nonalcoholic Fatty Liver Disease
Authors: KIM YOOMI , LEE DAEHO , LEE KIYOUNG
Published: June 2020

Aims/Introduction: Several non-invasive tools are available for the assessment of steatosis and fibrosis in nonalcoholic fatty liver disease (NAFLD) including clinical scoring systems, serum markers, transient elastography (TE), and magnetic resonance imaging (MRI) techniques, such as MRI-proton density fat fraction (MRI-PDFF) and MR elastography (MRE). In the present study, we aimed to evaluate whether MRI-based examinations better reflected the pathophysiology and metabolic features of NAFLD and outperformed other non-invasive methods.

Materials and Methods: A total of 120 subjects (31 healthy volunteers and 89 NAFLD patients) were subjected to anthropometric and laboratory tests, TE, and MRI-based examinations in this cross-sectional study, including liver biopsy in some patients (n=39).

Results: MRI-PDFF correlated with hepatic fat measured by MR spectroscopy (r=0.978, p<0.001) and was superior to the TE controlled attenuation parameter (CAP) (r=0.741, p<0.001). In addition, MRI-PDFF showed stronger correlations with various pathophysiologic parameters for cellular injury, glucose and lipid metabolism, and inflammation, than the CAP. The MRI-PDFF and CAP cutoff levels associated with abnormal elevation of serum alanine aminotransferase were 9.8% and 269 dB/m, respectively. The MRE liver stiffness measurement (LSM) showed stronger correlations with liver enzymes, platelets, complement component 3, several clinical fibrosis scoring systems, the enhanced liver fibrosis (ELF) score than the TE LSM. In an analysis of only biopsied patients, MRE and TE performed better in discriminating significant fibrosis (≥F2) than the ELF test.

Conclusions: Our results suggest that MRI-based assessment of NAFLD is the best non-invasive tool that captures the histologic, pathophysiologic and metabolic features of the disease.

Disclosure

Y. Kim: None. D. Lee: None. K. Lee: None.
1811-P: Induction of Hepatic Steatosis by a Methionine Choline Deficient Diet Promotes FGF21-Induced Insulin Secretion Independent of Diabetes
Authors: KIMURA TOMOHIKO , DE JESUS DARIO F. , FUKUDA KAZUKI , KAHRAMAN SEVIM , SHIBUE KIMITAKA , ORIME KAZUKI , KULKARNI ROHIT
Published: June 2020

While it is known that patients with nonalcoholic fatty liver disease exhibit enhanced insulin secretion, the factor(s) that contribute to this effect are not fully understood especially in nondiabetic subjects. Our aim was to examine crosstalk between a steatotic liver induced by a diet that is deficient in methionine choline (MCD) and pancreatic islets in the absence of diabetes/insulin resistance. Nine-week old male C57BL/6J mice were fed normal chow (CTL) or the MCD diet for 3 weeks followed by evaluation of glucose homeostasis, liver and pancreas pathology and hepatic gene expression by RNA-sequencing. Three weeks of MCD diet caused simple hepatic steatosis with hepatic triglyceride content 2-fold higher in MCD compared to CTL (n=5; p<0.05). Hepatic FGF-21 mRNA, protein and circulating FGF-21 levels were all significantly increased in MCD (129-fold, 8-fold, 5-fold respectively; n=5; p<0.05). Interestingly, while β-cell mass was not significantly altered between groups, insulin secretion as evaluated by ex-vivo GSIS in freshly isolated islets was significantly increased in MCD (5-fold; p<0.05, n=10). Examination of liver samples from patients with or without steatosis mimicked hepatic MCD data in the mouse model. Thus a significant increase in both mRNA and protein FGF-21 were evident in the steatotic group (3-fold, 5-fold respectively; n=6, p<0.05). Furthermore, 3 days treatment of nondiabetic human islets with recombinant FGF-21 protein led to a significant increase in GSIS compared to control (1.5-fold; p<0.05, n=4). FGF-21 treatment of EndoC-βH1 cells revealed significant activation of AKT and ERK1/2 signaling that likely promoted the enhanced insulin secretion. Taken together, liver steatosis significantly increases circulating FGF-21 to promote enhanced insulin secretion via improved signaling in nondiabetic beta cells.

Disclosure

T. Kimura: None. D.F. De Jesus: None. K. Fukuda: None. S. Kahraman: None. K. Shibue: None. K. Orime: None. R. Kulkarni: None.

Funding

National Institutes of Health (R01DK067536, R01DK103215)
182-LB: Cardiovascular Response to Exercise in T2D
Authors: ENGE DANIEL , REUSCH JANE E. , REGENSTEINER JUDITH G. , SCHÄFER MICHAL , HUNTER KENDALL S. , BARKER ALEX J. , NADEAU KRISTEN J. , SCALZO REBECCA L. , SCHAUER IRENE E. , ABUSHAMAT LAYLA , WHIPPLE MARY O. , RAFFERTY DEIRDRE
Published: June 2020

Type 2 diabetes (T2D) confers a greater likelihood of heart failure (HF) and hospitalization due to HF. Our group has reported sub-clinical cardiac dysfunction and aortic stiffness in youth and adults with uncomplicated T2D. As aortic stiffening is reported to precede HF and is common in people with uncomplicated T2D, we tested the hypothesis that aerobic exercise training will improve cardiac function and central arterial stiffness in people with T2D compared to participants of similar BMI without T2D. Sedentary adults with (n = 18, age 48±7, 45% female) and without (n = 20, age 45±7, 46% female) T2D, underwent MRI for cardiac function and aortic compliance (aortic relative area change [RAC] and pulse wave velocity [PWV]) before and after 15 weeks of aerobic exercise training. At baseline, both cohorts exhibit subclinical diastolic dysfunction, however, those with T2D had lower end-diastolic volume (EDVN,47.2±9.9 vs. 52.3±9.1 mL/m2) and end-systolic volume (ESVN,21.1±5.4 vs. 23.6±5.2 mL/m2), than controls. The T2D group had higher max thickness of the left ventricular (LV) septal wall (LVMTs, 10.5±1.8 vs. 9.3±1.6 mm) and LV lateral wall (LVMTl, 8.4±1.6 vs. 7.6±1.3 mm) than controls. Those with T2D also had lower RAC (12.7±1.6 vs. 20.2±1.6%) and higher PWV (7.4±1.3 vs. 3.9±0.6 m/s) than controls. Preliminary data in a subset post exercise intervention (n=7 with and n=8 without T2D), revealed increased EDVN(42.8±7.9 to 51.5±11.3 mL/m2) and ESVN(19.7±4.0 to 24.3±6.5 mL/m2) and decreased LVMTs(11.35±1.5 to 9.96±1.3 mm) and LVMTl(8.42±0.81 to 7.74±0.89 mm) in the T2D group, but cardiac endpoints were unchanged in controls. Post-intervention in both groups, ascending aorta RAC increased (control: 16.2±4 % to 18.5±1.7 and T2D: 10.6±1.7 % to 19.6±2.2 %) and PWV decreased (control: 6.3±2.0 to 3.4±0.3 and T2D: 8.0±1.4 to 2.9±0.7). Data from this study in progress suggest that adults with T2D have a greater degree of cardiac dysfunction and arterial stiffness than controls of similar BMI, which improve to a greater degree with exercise than in controls.

Disclosure

D. Enge: None. J.E. Reusch: None. J.G. Regensteiner: None. M. Schäfer: None. K.S. Hunter: None. A.J. Barker: None. K.J. Nadeau: None. R.L. Scalzo: None. I.E. Schauer: None. L. Abushamat: None. M.O. Whipple: None. D. Rafferty: None.

Funding

U.S. Department of Veterans Affairs (CX001532)
1820-P: A Direct AMPK Activator Reduces Liver Steatosis in a Mouse Model of NASH
Authors: MATHER KARLY M. , BOLAND MICHELLE L. , RIVERS EMMA , SRIVASTAVA ABHISHEK , SCHIMPL MARIANNE , ROBINSON JAMES , WAN PAUL T.C. , HANSEN JOSEFINE L. , READ JON A. , TREVASKIS JAMES L. , SMITH DAVID M.
Published: June 2020

Nonalcoholic fatty liver disease (NAFLD), is associated with metabolic disease and estimated to affect 25% of adults. NAFLD can progress to nonalcoholic steatohepatitis (NASH) which may lead to cirrhosis and hepatocellular carcinoma. There is no current treatment or cure for NASH.

AMPK (5’AMP-activated protein kinase) activators have shown potential for treating NAFLD/NASH due to their effects on fatty acid inhibition and cholesterol synthesis. To evaluate this potential, an AMPK activator (herein known as C455) was selected for a preclinical NASH study. C455 demonstrated potent and selective activation of AMPK (cellular EC50= 85 nM). X-ray crystallography showed the compound binds at the ADaM site, and pharmacokinetics determined a bioavailability of 45% and half-life of 3h.

Mice (male ob/ob on Amylin-NASH diet for 14-weeks or low-fat control) were dosed orally with vehicle or C455 at 3 or 30mg/kg daily. After 6 weeks of treatment, plasma ALT and terminal liver weight were decreased 32% and 22% respectively in mice treated with 30mg/kg C455 vs. vehicle. This dose also decreased liver lipid from 33% to 23%. Fibrosis via collagen staining did not improve with treatment, but significant decreases in mRNA transcripts for Col1a1(62%), Col1a2(59%), Col4a1(56%), and Timp1(55%) were seen with the high dose. Histology showed dose dependent increases in heart weight relative to body weight, and increased expression of genes associated with cardiac hypertrophy. Mice given 30mg/kg C455 also had increased cardiac glycogen storage compared to vehicle treated or LFD control mice.

Our study results show that systemic activation of AMPK in a diet-induced mouse model of NASH reduces plasma ALT, decreases liver lipid, and suppressed hepatic collagen gene expression but shows no histological improvement in fibrosis. Coupled with increased heart weight and glycogen storage, the data suggests that current small molecule activators of AMPK may not be viable therapies for the treatment of NASH-related fibrotic disease.

Disclosure

K.M. Mather: None. M.L. Boland: None. E. Rivers: None. A. Srivastava: None. M. Schimpl: Employee; Self; AstraZeneca. Stock/Shareholder; Self; AstraZeneca. J. Robinson: None. P.T.C. Wan: None. J.L. Hansen: None. J.A. Read: Employee; Self; AstraZeneca. Stock/Shareholder; Self; AstraZeneca. J.L. Trevaskis: Employee; Self; Gilead Sciences, Inc. D.M. Smith: Employee; Self; AstraZeneca.
1821-P: Foz Foz Mice with a Mutation in the Alms1 Gene Model the Metabolic Syndrome with Advanced Stages of Nonalcoholic Steatohepatitis and Fibrosis
Authors: LIU JIANYING , WANG SHENG-PING , D’AQUINO KATHARINE E. , HO GEORGE , WALLACE NATHANIEL H. , RANKIN MATTHEW M. , DU FUYONG , HINKE SIMON A. , ALBARAZANJI KAMAL A. , NAWROCKI ANDREA R.
Published: June 2020

Nonalcoholic fatty liver disease (NAFLD) and its progression to nonalcoholic steatohepatitis (NASH) is highly prevalent in modern societies frequently consuming diets rich in fats and carbohydrates. NASH is characterized by liver steatosis, inflammation and increased risk for liver fibrosis in human. There is no single animal model available that encompasses the defining features of NASH; therefore, it is of importance to thoroughly characterize available rodent models for NASH research and new drug discovery. Foz/foz mice have a mutation in the Alms1 gene, a ubiquitous protein essential for proper primary cilium function. They have similar defects such as metabolic syndrome, NASH and liver fibrosis to human with the same gene mutation. To better characterize the model, foz/foz mice were fed with western diet (WD, D12079B) for various duration. Compared with WT mice, foz/foz mice on WD had (1) increased BW gain driven by hyperphagia; (2) insulin/leptin resistance with decreased adiponectin levels and hyperlipidemia; (3) increased fat mass and hepatomegaly; (4) increased liver enzymes and elevated TIMP1 in plasma; (5) Liver gene expressions profiles of foz/foz mice showed up-regulation of fibrogenic genes and Lipogenesis, inflammation genes. Histopathology assessment showed foz/foz mice developed severe hepatic steatosis and increased marker of macrophage populations (F4/80 IHC staining). liver collagen content was significantly increased at 16 and 24 wks post WD feeding (PSR staining) compared to wild type mice fed the same diet. Our results suggest that the foz/foz mouse model has robust hallmark features of NASH such as metabolic syndrome, steatohepatitis, and fibrosis. In addition, our data suggest the presence of metabolic complications in other organs as well, as foz/foz mice develop proteinuria over time. The WD-induced foz/foz model exhibits several clinically relevant features and serves as a translational model of NASH.

Disclosure

J. Liu: None. S. Wang: None. K.E. D’Aquino: Employee; Self; Janssen Pharmaceuticals, Inc. Employee; Spouse/Partner; Janssen Pharmaceuticals, Inc. N.H. Wallace: None. M.M. Rankin: Employee; Self; Janssen Pharmaceuticals, Inc. Stock/Shareholder; Self; Johnson & Johnson. F. Du: None. S.A. Hinke: Employee; Self; Janssen Pharmaceuticals, Inc. Stock/Shareholder; Self; Johnson & Johnson. K.A. Albarazanji: None. A.R. Nawrocki: Employee; Self; Janssen Pharmaceuticals, Inc.
1828-P: Diurnal Pattern of Hepatic Insulin Extraction in Healthy and Type 2 Diabetes
Authors: ROMERES DAVIDE , YADAV YOGESH R. , RUCHI FNU , COBELLI CLAUDIO , MAN CHIARA DALLA , BASU ANANDA , BASU RITA
Published: June 2020

Hepatic extraction of insulin is associated with insulin sensitivity in humans. It is unknown if a diurnal pattern of HE exists and if this differs between nondiabetic (ND) and type 2 diabetes (T2D) subjects. Here are data presented for 10 ND (BMI 29.4±1.3 kg/m2, age 57±3 yrs) and 9 T2D subjects (BMI 33±2 kg/m2, age 58±4 yrs, HbA1c 57±4 mM/mol) studied thus far. They ingested identical mixed meals (75 g carbs; 8 kcal/kg) at either breakfast (B), lunch (L) or dinner (D) in randomized Latin square order on three consecutive days with frequent sampling of plasma glucose (G), insulin (I) and C-peptide (C) concentrations. Fasting and post-meal glucose concentrations were higher while fasting and post-meal insulin, C-peptide concentrations were lower during meals in T2D as compared to ND subjects. Total and basal hepatic insulin extraction (HE and HEb, respectively) were estimated using the Oral Insulin and C-peptide Minimal Models. Differences with B, L and D and between ND and T2D were assessed by N-way ANOVA (a p-value<0.05 was considered statistically significant). Results show that both HE and HEb were significantly lower (p=0.009, p=0.011, respectively) throughout the day in T2D than ND subjects (Fig 1). Our initial results suggest an emerging diurnal pattern of HE in T2D subjects with D being lower than other meals. These observations provide novel insights into the diurnal pattern of hepatic extraction throughout the day in T2D subjects.

Disclosure

D. Romeres: None. Y.R. Yadav: None. F. Ruchi: None. C. Cobelli: None. C. Dalla Man: Research Support; Self; Sanofi-Aventis Deutschland GmbH. A. Basu: Consultant; Spouse/Partner; GENFIT. Research Support; Spouse/Partner; AstraZeneca. R. Basu: Consultant; Self; GENFIT. Research Support; Self; AstraZeneca.

Funding

National Institutes of Health (R01DK029953 to R.B.), (R01DK085516 to A.B.), (DK059637, DK020593)
1832-P: Palmitic Acid Esters of Hydroxy Stearic Acids Reduce Endogenous Glucose Production through GPR43 Activation
Authors: SANTORO ANNA , ZHOU PENG , ZHU YAN , PERONI ODILE D. , NELSON ANDREW T. , SIEGEL DIONICIO , KAHN BARBARA
Published: June 2020

Palmitic Acid esters of Hydroxy Stearic Acids (PAHSAs), bioactive lipids with anti-inflammatory and antidiabetic effects, improve systemic and hepatic insulin sensitivity in insulin-resistant mice. PAHSAs augment insulin action on endogenous glucose production (EGP) through direct and indirect actions involving inter-tissue communication between liver and adipose tissue. The direct effect of PAHSAs to inhibit EGP is mediated through a cAMP dependent pathway involving Gα/i protein-coupled receptors. Here, we investigated which receptor mediates PAHSA effects on EGP. We found that 9-PAHSA activates GPR43 in a Ca+2 flux assay. 4-CMTB (0.4μM), a selective agonist-allosteric modulator of GPR43, reduced basal EGP by 23% and glucagon-stimulated EGP by 20% in primary hepatocytes ex vivo. Acute 9-PAHSA treatment (40μM) inhibited basal EGP to the same extent as 4-CMTB. 9-PAHSA suppression of basal EGP was abolished in primary hepatocytes from whole body GPR43 KO mice. Intravenous infusion of 9-PAHSA (9μg/hr) for 3 hrs decreased ambient EGP in chow-fed mice studied 5 hours after food removal (WT: vehicle= 24.2±1.9 vs. PAHSA= 17.2±0.7 mg/kg/min p<0.05). This effect was absent in GPR43 KO mice (KO: vehicle= 25.3±4.2 vs. PAHSA= 23.4±2.7 mg/kg/min). We also examined whether GPR43 activation is required for the direct effect of 9-PAHSA on WAT lipolysis. Treatment with the β-adrenergic agonist isoproterenol increased FFA release from WAT explants by 3.8-fold, and 9-PAHSA inhibited isoproterenol-induced lipolysis by 21% which was similar to the effects of insulin and 4-CMTB. In WAT explants from GPR43 KO mice, 9-PAHSA suppression of isoproterenol-induced FFA release was abolished while insulin still suppressed lipolysis.

Sum: 9-PAHSA activates GPR43, which is involved in its direct effects to reduce hepatic glucose production and WAT lipolysis.

Conclusion: We have identified a new receptor that mediates some of the beneficial metabolic effects of PAHSAs.

Disclosure

A. Santoro: None. P. Zhou: None. Y. Zhu: None. O.D. Peroni: None. A.T. Nelson: None. D. Siegel: None. B. Kahn: Advisory Panel; Self; Harrington Discovery Institute, Janssen Pharmaceuticals, Inc., National Institute of Diabetes and Digestive and Kidney Diseases.

Funding

American Diabetes Association (1-18-PDF-134 to P.Z.); National Institutes of Health (DK43051, DK57521, DK106210, DK112622); JPB Foundation; American Heart Association
1833-P: A Novel Genetic Modified and Pegylated FGF21 Analog for the Treatment of Nonalcoholic Steatohepatitis in Nonhuman Primates
Authors: CUI AOYUAN , LI JIAN , JI SHAOHUI , MA FENGGUANG , WANG GENBEI , XUE YAQIAN , LIU ZHENGSHUAI , MA XIAOHUI , LI YU
Published: June 2020

Nonalcoholic steatohepatitis (NASH) is becoming a major cause of liver diseases with no effective drugs in the clinic. Fibroblast growth factor 21 (FGF-21) has emerged as a potent metabolic regulator with therapeutic prospect to lipids and NASH in animal models and preclinical trials. FGF-21 and the agents mimicking its activity may be developed into the pharmacotherapeutic targets for NASH treatment. Here, we show that B1344, a site-specific PEGylated human FGF-21 analog, shifts physiology of NASH mice or cynomolgus monkeys towards that of the control animals fed with standard diet, and significantly improves their NASH phenotypes. To determine the role of B1344 in the progression of murine NAFLD independent of obesity, methionine and choline deficiency (MCD) diet-fed mouse model of NASH were generated. Both short- and long-term administration of B1344 with the doses of 0.125 mg/kg and 0.2 mg/kg in mice protect against lipotoxicity damage and liver fibrosis. Expression levels of proinflammatory and fibrotic genes are decreased in livers of mice treated with B1344. Moreover, pharmacologic efficacies of B1344 on NASH were evaluated in NAFLD cynomolgus monkeys fed with high-fat diet. Strikingly, administration of B1344 causes a potent reduction of hepatic steatosis and fibrosis as measured by magnetic resonance imaging proton density fat fraction (MRI-PDFF) and Sirius Red staining, and protects the monkeys from the progression of steatohepatitis. In addition, administration of B1344 results in a reduction of body weight and adiposity, and improvement of lipid profile and glycemic control. The biological properties of the novel FGF-21 analog B1344 is identical to human FGF-21. Together, these demonstrate impressive effects of B1344 on NASH and metabolic parameters, and may represent novel therapeutics for treating these diseases in humans.

Disclosure

A. Cui: None. J. Li: None. S. Ji: None. F. Ma: None. G. Wang: None. Y. Xue: None. Z. Liu: None. X. Ma: None. Y. Li: None.

Funding

Tasly Pharmaceutical Co. Ltd.; National Key Research and Development Program of China (2019YFA0802502); National Natural Science Foundation of China (81925008); Chinese Academy of Sciences (XDA12030210); Shanghai Science and Technology Commission (19140903300); National Science and Technology Major Projects (2019ZX09201001-001-001)
184-OR: Liver Fibrosis Scores Do Not Predict Response to Therapy in T2DM
Authors: LAVRYNENKO OLGA , ALATRACH MARIAM , ABDUL-GHANI MUHAMMAD , CERSOSIMO EUGENIO , ALKHOURI NAIM , PUCKETT CURTISS L. , DEFRONZO RALPH A. , ADAMS JOHN M. , VASQUEZ JUAN A. , CLARKE GEOFFREY D.
Published: June 2020

Patients with NAFLD and T2DM are at high risk of liver fibrosis. Pioglitazone(Pio) and GLP-1RAs have shown efficacy against NAFLD. The EDICT trial assessed Triple (pio/exenatide/metformin(met)) vs. Conventional(Conv) (met/SU/insulin) Therapy.

Aim: Evaluate the effect of these approaches on liver fibrosis scores and hepatic fibrosis, steatosis (FibroScan).

Methods: 144 newly diagnosed T2DM were randomized to receive Triple or Conv Therapy to maintain HbA1c <6.5%. After 2 yrs baseline measurements were repeated and liver fibrosis scores (AST/ALT ratio, APRI, FIB-4, NFS) were calculated.

Results: Patients were well matched for age, BMI, HbA1c (8.8%), LFTs. Neither therapy reduced any liver fibrosis score. Triple, but not Conv, reduced AST/ALT (p<0.001). The greatest AST/ALT reductions with Triple Therapy occurred in subjects in highest AST/ALT tertile at baseline. Only 1 subject receiving Triple Therapy had fibrosis score >0, while 43% of Conv Therapy subjects had fibrosis score of F3/F4 (p<0.0001). 87% of Conv Therapy subjects had steatosis score of S2/S3 vs. to 38% of Triple Therapy (p<0.001) (Table).

Conclusion: Both Triple (6.0%) and Conventional Therapy (6.7%) markedly reduced HbA1c after 2 yrs, but only Triple Therapy reduced AST/ALT. Fibrosis scores did not change in either group and are not useful in predicting response to therapy. Triple Therapy completely prevented fibrosis and reduced steatosis by >50 % vs. Conventional Therapy.

Disclosure

O. Lavrynenko: None. M. Alatrach: None. M. Abdul-Ghani: None. E. Cersosimo: None. N. Alkhouri: Research Support; Self; Allergan plc. C.L. Puckett: None. R.A. DeFronzo: None. J.M. Adams: None. J.A. Vasquez: None. G.D. Clarke: None.