bluuebunny/crossref_metadata_2025 huggingface dataset:
1841-P: Fasting Test Distinguished Organic from Functional Hypoglycemia
Authors: SKRHA JAN , PRAZNY MARTIN , KRIZOVA JARMILA , SKRHA JAN
Published: June 2019

Nondiabetic hypoglycemia may be caused by hepatic or renal failure, drugs, malnutrition, malignancy or functional changes. The fasting test may be used to distinguish organic hypoglycemia by insulinoma from functional (reactive) hypoglycemia. The aim was to compare if clinical and biochemical characteristics could clearly differentiate both diagnoses during fasting test.

In total, 84 patients (mean age 50 ± 16 years; 54 females) with history of unclear hypoglycemia were examined at our department in the last 8 years. Their liver and renal functions were normal. All patients underwent a 72-hour fasting test. The test was stopped earlier when neuroglycopenic symptoms developed. Blood samples were taken for glucose (BG), insulin (Ins) and C peptide measurements at the beginning, in 6-hour intervals and at the end of the test.

Fasting test was positive (F+ group) in 43 persons developing neuroglycopenic symptoms and thus it was stopped earlier (duration 24 ± 17 hours) as compared to F- group (41 persons) without these symptoms during the whole 72-hours of fasting. Both groups were similar in age, BMI or arterial pressure. Significant differences between F+ and F- group were observed at the end of the test (BG: 2.0 ± 0.4 vs. 3.8 ± 0.8 mmol/l, p<0.0001; Ins: 61 (11; 111) vs. 13 (5; 22) mU/L, p<0.0001; C peptide: 1.4 (0.5; 2.1) vs. 0.4 (0.1; 0.6) nmol/L, p<0.0001). All patients in F+ group developed spontaneous hypoglycemia <2.5 mmol/l during fasting but it was not true in F- group. The insulin/BG ratio (normal value <5.0 mU/mmol) could significantly distinguish both groups at the end of fasting (29.7 vs. 3.5 mU/mmol, p<0.0001). Insulinoma was then confirmed in all F+ patients at surgery.

Fasting test represents the most important diagnostic tool in the evaluation of unclear nondiabetic hypoglycemia. Neuroglycopenic symptoms together with spontaneous hypoglycemia by fasting are the most sensitive indicators of insulinoma. In addition, the insulin/BG ratio seems to be of highest benefit in distinguishing organic and functional hypoglycemia.

Disclosure

J. Skrha: Advisory Panel; Self; Boehringer Ingelheim International GmbH. Speaker's Bureau; Self; Lilly Diabetes. M. Prazny: Advisory Panel; Self; Abbott, AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Medtronic, Merck Sharp & Dohme Corp., Novo Nordisk Inc., Sanofi-Aventis, Takeda Pharmaceutical Company Limited. Board Member; Self; Bayer AG. Consultant; Self; Medtronic, Roche Diabetes Care. Speaker's Bureau; Self; Abbott, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Medtronic, Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Sanofi-Aventis, Servier, Teva Pharmaceutical Industries Ltd. J. Krizova: None. J. Skrha: Advisory Panel; Self; Boehringer Ingelheim International GmbH, Lilly Diabetes, Novo Nordisk A/S, Sanofi.

Funding

RVO-VFN00064165
1856-P: A High Sugar (Fructose) Diet Increases C-Reactive Protein (CRP) in Rhesus Macaques: Relationships with Changes of Apolipoproteins and Adiponectin
Authors: HAVEL PETER J. , GRAHAM JAMES L. , STANHOPE KIMBER L. , BUTLER ANDREW
Published: June 2019

CRP is produced by the liver in response to factors released from macrophages and adipocytes. High circulating CRP concentrations are therefore an indicator of inflammation and/or injury. We examined the effects of dietary fructose on plasma CRP concentrations in a nonhuman primate model of diet-induced metabolic syndrome. Adult male rhesus macaques 12±3 years of age, weighing 16.2±1.9 kg (n=29) maintained on chow (LabDiet 5047) ad libitum were provided with flavored fructose beverages containing 75g of fructose (300 kcal/day). Fasting plasma samples were collected at baseline and after 1 and 3 months. Indices of glucose homeostasis (insulin, glucose, HOMA-IR), adiposity/metabolic homeostasis (leptin, adiponectin) and lipid metabolism (TG, cholesterol, LDL-C, HDL-C, VLDL-C, ApoA1, ApoB, ApoC3 and ApoE) were analyzed. Fructose consumption induced weight gain +1.5±0.2 kg, and insulin resistance, accompanied by increases of plasma leptin, decreased adiponectin, hypertriglyceridemia and increases of ApoC3 and ApoE. Plasma CRP increased from 1.9±0.2 mg/l to 2.9±0.3 mg/l (%∆ = 81±13%) and 2.9±0.3 mg/l (%∆ = +91±14%; both p<0.001) at 1 and 3 months respectively. Bivariate correlation analysis of log-transformed data compared fructose-induced changes (Δ) of CRP with Δ’s of body weight and metabolic indices. After 1 month of fructose consumption, strong positive associations were identified between ΔCRP and ΔApoE (r=0.62, p<0.001), ΔApoC3 (r=0.56, p=0.002), and negatively with Δadiponectin (r=-0.49, p=0.007). After 3 months of fructose exposure, the strong associations between ΔCRP and ΔApoE (r=0.49, p=0.007) and significant relationships with ΔApoC3 (r=0.45, p=0.015) and Δadiponectin (r=-0.39, p=0.038) remained. These results support an important link between fructose diet-induced inflammation with changes of the anti-inflammatory adipocyte hormone (adiponectin) and dysregulated hepatic lipoprotein metabolism.

Disclosure

P.J. Havel: Research Support; Self; Arrowhead Research, Bristol-Myers Squibb Company, Magnamosis Inc. J.L. Graham: Research Support; Self; Arrowhead Research, Bristol-Myers Squibb Company, Magnamosis Inc. K.L. Stanhope: None. A. Butler: None.

Funding

National Institutes of Health
1857-P: Modified Mesenchymal Stromal Cells: A Novel and Safe Therapy for Treating Nonalcoholic Fatty Liver Disease (NAFLD) in Obesity and Diabetes
Authors: DOMINGUES CLEYTON C. , KUNDU NABANITA , KROPOTOVA YANA , SEN SABYASACHI
Published: June 2019

Background: Mesenchymal stromal cells (MSCs) can home-in to inflamed fat depots to deliver antioxidants locally and help reduce oxidative stress, inflammation and improve NAFLD in high-fat diet (HFD) induced obese (DIO) diabetic mice.

Methods: Antioxidants Sod2 (mitochondrial), Catalase (cytosolic) or Null (control) genes were upregulated individually, or in combination in adipose-derived MSCs. Modified MSCs were examined in-vitro in presence of adipogenic media to mimic DIO milieu. Next, modified MSCs were delivered intraperitoneally (IP) in mice that received 45% or 60% HFD for 8 to16 weeks.

Results: In-vitro, we noted reduced MSC lipid droplets with SOD2, Catalase and combination compared to Null. Inflammatory marker IL6 mRNA, was down-regulated in SOD2, Cat and combination MSCs vs. Null-MSCs. In-vivo, n=4, Glucose tolerance test (GTT) improved with SOD2 MSC delivery in (p=0.07) at week-4. Plasma inflammatory marker TNFa, was reduced in both SOD2 and Catalase MSC groups (p<0.05) vs. Null-MSCs group. Analysis of omental and pericardial fat showed significant up-regulation in mRNA expression of brown fat marker, Ucp1 (∼1000 and 100-fold, respectively) and PGC1A mRNA was upregulated. Increased Ucp1 was confirmed by staining. There was a reduction in liver fat content by histology and liver triglyceride assay (p<0.05). Outcome measures of SOD2+Catalase combination MSC delivery in-vivo is pending.

Conclusion: Upregulation of SOD2 plus Catalase, in-vitro, reduced inflammation in adipogenic media more so than individual gene upregulation. In-vivo, delivery of Sod2 and Catalase upregulated individually in MSCs, reduced inflammation and improved GTT with concomitant increased browning of white fat and reduced liver fat. Our results indicate that antioxidant upregulated modified MSC delivery IP, to target inflamed fat depots helps reduce oxidative stress and inflammation and can be an efficient tool for treating obesity, diabetes and NAFLD.

Disclosure

C.C. Domingues: None. N. Kundu: None. Y. Kropotova: None. S. Sen: None.

Funding

Clinical and Translational Science Institute at Children's National
1862-P: Insulin-Independent Glucose Utilization during Exercise Is Impaired in Type 1 Diabetes: A New Model Based Analysis
Authors: ROMERES DAVIDE , SCHIAVON MICHELE , BASU ANANDA , COBELLI CLAUDIO , BASU RITA , MAN CHIARA DALLA
Published: June 2019

We have previously shown that low insulin coupled with euglycemia or mild hyperglycemia appear to be the most favorable milieu for type 1 diabetes (T1D) during exercise. However, the contributions of insulin-dependent (IDGU) and insulin independent (IIGU) effects of exercise on glucose utilization are unknown. To do so, we studied 6 T1D (age 29±3 years, BMI 28.3±2.0 kg/m2, VO2 max 28.7±2.9 ml/kg/min, mean±SE) and 6 ND (age=28.2±4.2 year, BMI=23.6 ±1.0 kg/m2, VO2 max 36.6±2.9 ml/kg/min) subjects with a glucose clamp, during, before and after a 60 min exercise session at 65% VO2max, on three visits: euglycemia-low insulin (V1); euglycemia-high insulin (V2) and hyperglycemia-low insulin (V3) in random order. Glucose turnover was measured with the isotope dilution technique using [6,6-2H2]glucose. We modified the minimal model to measure the IDGU and IIGU components of glucose utilization (Figure 1, upper panel). Results demonstrate that while IDGU was similar in both groups (data not shown), IIGU (expressed as percent change from baseline) during exercise was significantly lower in T1D than ND subjects, though the trend across the visits were similar (Figure 1, lower panel). Further studies are necessary to determine the cause for this abnormality in T1D to optimize glucose control.

Disclosure

D. Romeres: None. M. Schiavon: None. A. Basu: Research Support; Spouse/Partner; AstraZeneca. C. Cobelli: None. R. Basu: Other Relationship; Self; AstraZeneca. C. Dalla Man: None.

Funding

National Institutes of Health (R01090541)
1871-P: Mir 103/107 Regulates Lipid Metabolism by Inhibiting Betatrophin/ANGPTL8
Authors: ABU-FARHA MOHAMED , CHERIAN PREETHI THOMAS , KHAIRI IRINA AL , AL-MULLA FAHD , ABUBAKER JEHAD
Published: June 2019

Angiopoietin-like protein 8 (ANGPTL8)/betatrophin is liver and adipocyte produced hormone that has been associated with insulin resistance and dyslipidaemia. The interaction of ANGPTL8 with ANGPTL3 causes an inhibition of lipoprotein lipase (LPL) activity and ultimately increases serum levels of triglyceride. Identification of ANGPTL8 and ANGPTL3 inhibitors have been pursued as potential drug target to treat dyslipidaemia. In this study we used target prediction algorithms to show that mir103 has a putative binding sites in the 3' untranslated region of ANGPTL8. Overexpression of mir103 in HepG2 cells or its mimic resulted in decreased ANGPTL8 RNA and protein level compared to scrambled microRNA control. Cloning the 3’UTR of ANGPTL8 with Luciferase reporter assay confirmed the direct binding and repression of luciferase activity by mir103 to ANGPTL8 compared to its control. Palmitic acid treatment of HepG2 cells mimicked dyslipidaemia and increased ANGPTL8 level which was reduced after treatment with mir103. Additionally, expression of miR103 and ANGPTL8 in circulation and adipose tissue was evaluated in 144 participants that included 82 non-obese (BMI <30 kg/m2) and 62 obese (BMI >30 kg/m2). Mir103 levels in small RNA extracted from plasma and adipose tissue was measured by RT-PCR. Mir103 was higher in non-obese people in both plasma and adipose tissue. ANGPTL8 level on the other hand was higher in obese individuals (1200.5 ±110.2 pg/ml) compared to non-obese (720.7 ±80.9 pg/ml, p-value =0007). MiR103 was inversely associated with BMI (p-value < 0.001), leptin (p-value = 0.010) and triglyceride (p-value =0.011). Taken together, our data confirms the binding and inhibition of mir103 to ANGPTL8 under normal and high fat conditions. It also sheds light on the potential role of mir103 in regulating lipid metabolism which can be used as a future therapeutic target for dyslipidaemia and other metabolic related diseases.

Disclosure

M. Abu-Farha: None. P.T. Cherian: None. I. Al Khairi: None. F. Al-Mulla: None. J. Abubaker: None.

Funding

Dasman Diabetes Institute; Kuwait Foundation for the Advancement of Sciences (RA2016-025)
188-OR: Partial Depletion of CD206 M2-Like Macrophages Enhances Preadipocyte Differentiation via Reduction of Wnt Signaling
Authors: NAWAZ ALLAH , NAKAGAWA TAKASHI , YAGI KUNIMASA , FUJISAKA SHIHO , TOBE KAZUYUKI , OKABE KEISUKE , KADO TOMONOBU , LIU JIANHUI , BILAL MUHAMMAD
Published: June 2019

Several pieces of evidence indicate that Wnt/β-catenin pathway may regulate the mesenchymal stem cells fate proliferation in vivo. The Wnt signaling is also known to inhibit preadipocyte differentiation into adipocyte. Adipose tissue macrophages play important roles in the maintenance of tissue homeostasis by regulating insulin sensitivity via their anti-inflammatory actions. However, how CD206 M2-like macrophages linked to Wnt signaling in maintaining the preadipocyte differentiation, remains elusive. In the present study, we aimed to investigate the physiological role of CD206 linked to Wnt signaling in preadipocyte differentiation. We show that C1q, which can mediate Wnt signaling, is abundantly expressed in CD206 M2-like macrophages in epididymal adipose tissue. Then, we examined whether CD206 M2-macrophages depletion activate or inhibit Wnt signaling and preadipocyte differentiation. Here, we show that the expression of M2 marker genes was reduced with a reduced expression of Wnt signaling genes in adipose tissue of CD206-reduced mice. Western blot analysis confirmed reduced protein level of β-catenin in CD206-reduced mice that can contribute to regulate the preadipocyte/progenitor’s activity. We presumed that the reduction of C1q-induced Wnt signaling promoted adipogenesis. We also found that depletion of CD206 M2-like macrophages resulted in the generation of the smaller adipocyte, upregulated expression of metabolically favorable genes and enhanced insulin sensitivity in both chow and high-fat diet-fed CD206-reduced mice. Taken together, we show that partial depletion of CD206 M2-like macrophages inhibits Wnt/β-catenin pathway activity in mice and increases preadipocyte differentiation, thus ameliorating insulin resistance in lean and obese mice.

Disclosure

A. Nawaz: Research Support; Spouse/Partner; Astellas Pharma Inc., Astellas Pharma Inc., AstraZeneca, Daiichi Sankyo Company, Limited, Eli Lilly and Company. Research Support; Self; Japan Society for the Promotion of Science. Research Support; Spouse/Partner; Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Novo Nordisk A/S, Ono Pharmaceutical Co., Ltd., Pfizer Inc., Sanofi, Shionogi & Co., Ltd., Takeda Pharmaceutical Company Limited. T. Nakagawa: None. K. Yagi: None. S. Fujisaka: None. K. Tobe: Advisory Panel; Self; Novo Nordisk A/S. Board Member; Self; Japan Diabetes Society. Research Support; Self; Bristol-Myers Squibb Company. K. Okabe: None. T. Kado: None. J. Liu: None. M. Bilal: None.

Funding

Japan Society for the Promotion of Science
1881-P: The SGLT2 Inhibitor Dapagliflozin Improves Glucose Control and Enhances Liver Fatty Acid Oxidation and Ketone Body Formation in Insulin Resistant Obese Zucker Rats
Authors: KROON TOBIAS , HAGSTEDT THERESE , KJELLSTEDT ANN , LINDBLOM ANNA , LÖFGREN LARS , BOUCHER JEREMIE , SÖRHEDE WINZELL MARIA , LINDEN DANIEL , WALLENIUS KRISTINA
Published: June 2019

Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are effective treatments for improving glucose control and reducing cardiovascular events in patients with T2D. In the present study we examined the metabolic effects of chronic treatment of the SGLT2i dapagliflozin in obese insulin resistant Zucker rats. Three different studies were performed with the aims to assess: 1) liver CoA intermediates, 2) whole-body FA metabolism using a constant infusion of 3H-palmitate and 3) tissue-specific rates of FA utilization and storage using 3H-labelled bromo-palmitate and 14C-labelled palmitate, respectively. Four weeks treatment with dapagliflozin (1 mg/kg/d p.o.) markedly increased glucosuria, reduced HbA1c and fasting plasma glucose and insulin levels and increased pancreatic insulin content compared to vehicle treated rats. Liver acetyl-CoA levels were increased while the malonyl-CoA/acetyl-CoA ratio was decreased following dapagliflozin treatment, suggestive of an increased liver FA oxidation. Treatment with dapagliflozin also increased liver HMG-CoA levels, an intermediate in ketone body formation, indicating that the acetyl-CoA is diverted to ketone body formation. This was supported by increased fasting plasma levels of the ketone body β-hydroxybutyrate in the dapagliflozin-treated rats compared to vehicle. Dapagliflozin treatment resulted in increased plasma FA levels and rates of whole-body FA oxidation. The increased whole-body FA oxidation was subsequently found to be driven by an increase in liver FA utilization.

In conclusion, dapagliflozin improves glucose homeostasis and enhances FA utilization in insulin resistant obese animals. These results suggest that dapagliflozin-induced glucose loss in urine signals a fasting state to the liver, which leads to an increased FA oxidation and ketone body production.

Disclosure

T. Kroon: Employee; Self; AstraZeneca. T. Hagstedt: None. A. Kjellstedt: None. A. Lindblom: Employee; Self; AstraZeneca. L. Löfgren: Employee; Self; AstraZeneca. J. Boucher: Employee; Self; AstraZeneca. M. Sörhede Winzell: Employee; Self; AstraZeneca. D. Linden: Employee; Self; AstraZeneca. K. Wallenius: Employee; Self; AstraZeneca. Stock/Shareholder; Self; AstraZeneca.
1898-P: The Role of the Diabetes Gene and Wnt Pathway Effector TCF7L2 in Hepatic Metabolic Zonation
Authors: AYALA IRISCILLA I. , DEFRONZO RALPH A. , NORTON LUKE
Published: June 2019

The Transcription Factor 7-like 2 (TCF7L2) gene is associated with type 2 diabetes (T2D), but the function of the protein in metabolic tissues is controversial. The liver regulates whole body lipid and carbohydrate metabolism, and the development of T2D and nonalcoholic fatty liver disease (NAFLD) is linked to hepatic insulin resistance and dyslipidemia. The liver is metabolically zonated and hepatocytes are specialized to execute the diverse metabolic functions of the liver. We previously demonstrated that TCF7L2 transcriptionally regulates multiple metabolic pathways in hepatocytes, and hypothesized that TCF7L2 controls metabolic zonation in vivo. We generated mice expressing inactive TCF7L2 in liver (Hep-ΔE11-TCF7L2) and examined zonation, lipid metabolism and the development of NAFLD. We observed complete disruption of hepatic zonation in Hep-ΔE11-TCF7L2, as determined by the absence of glutamine synthetase expression in pericentral (PC) hepatocytes. Interestingly, using laser capture microdissection and TCF/LEF reporter mice, we observed that TCF7L2 and Wnt/β-catenin signaling genes were expressed throughout the liver, but that the activity of TCF7L2 was restricted to PC hepatocytes. Consistent with TCF7L2 activity in glycolytic PC hepatocytes only, Hep-ΔE11-TCF7L2 mice had normal glucose tolerance and insulin sensitivity. However, HFD-fed Hep-ΔE11-TCF7L2 mice were protected from adipose tissue expansion and weight gain, but had elevated circulating FFA. In a subset of Hep-ΔE11-TCF7 mice, we detected a severe NASH phenotype histologically, and RNA-Seq analysis revealed significant alterations in the expression of genes involved in cholesterol and lipid metabolism, and inflammatory and fibrotic pathways. These data show that TCF7L2 activity is highly restricted in mature liver, but that TCF7L2 is required for hepatic metabolic zonation. Our data also suggest that disrupted metabolic zonation may contribute to the development of NAFLD.

Disclosure

I.I. Ayala: None. R.A. DeFronzo: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Elcelyx Therapeutics, Inc., Intarcia Therapeutics, Inc., Janssen Pharmaceuticals, Inc., Novo Nordisk Inc. Research Support; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Pharmaceuticals, Inc., Merck & Co., Inc. Speaker's Bureau; Self; AstraZeneca, Novo Nordisk Inc. L. Norton: None.

Funding

National Institutes of Health
1902-P: Definition of Nonalcoholic Fatty Liver Disease (NAFLD) Stage by Independent Noninvasive Fibrosis Markers and Elastography
Authors: LAVRYNENKO OLGA , ZEMLIANITSYNA OLHA , KRAVCHUN NONNA , DUNAEVA INNA , KARACHENTSEV IURIY , SINAIKO VADYM , DEFRONZO RALPH A. , CERSOSIMO EUGENIO
Published: June 2019

Aim: We sought to define the degree of NAFLD with B-mode ultrasound sono-elastography using the discriminant Bonacini scale in type 2 diabetes (T2D) patients.

Materials and Methods: Forty-three patients with T2D and clinically diagnosed NAFLD (26F, 17M) were examined with liver ultrasound in B-mode elastography. The presence and degree of fibrosis was assessed using the Bonacini Classification (Discriminant) Scale. The compliance of the stage of the fibrosis was compared with results of the histological evaluation of METAVIR and Ishak.

Results: An enlarged liver was found in all, often combined with the hyperechogenic, coarse-grained structure. In rare cases, a decrease in the liver echogenicity indicated development of fibrosis. The liver density index was significantly higher in men (2.63 ± 0.05) than in women (2.49 ± 0.06, p = 0.05). In a series of correlation analyses, it was found that sono-elastography strongly correlated with the results of the classification (discriminant) Bonacini score scale (r= 0.68 ± 0.13, p <0.001). The results of the Ishak and METAVIR scales at minimum values have a moderate correlation with sono-elastography data (r= 0.36 ± 0.17, p < 0.05 and r= 0.37±0.16, p<0.05respectively). These scales determine the degree of fibrosis as a range from the less to the greater degree, but the stronger correlation (r= 0.50 ± 0.16, p<0.01and r= 0.47 ± 0.15, p<0.01respectively) with results of sono-elastography was found in patients with severe fibrosis of the liver.

Conclusion: In majority of type 2 DM patients with clinical NAFLD a slight degree fibrosis (F0-F2) was diagnosed. To better assess the degree of liver fibrosis in these patients it is advisable to use sono-elastography and the discriminant Bonacini scale. These have similar significance, can complement each other and allow identification of early manifestations of liver fibrosis in patients with type 2 DM.

Disclosure

O. Lavrynenko: None. O. Zemlianitsyna: None. N. Kravchun: None. I. Dunaeva: None. I. Karachentsev: None. V. Sinaiko: None. R.A. DeFronzo: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Elcelyx Therapeutics, Inc., Intarcia Therapeutics, Inc., Janssen Pharmaceuticals, Inc., Novo Nordisk Inc. Research Support; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Pharmaceuticals, Inc., Merck & Co., Inc. Speaker's Bureau; Self; AstraZeneca, Novo Nordisk Inc. E. Cersosimo: None.
191-OR: New Regulators of Insulin-Stimulated GLUT4 Exocytosis
Authors: SHEN JINGSHI
Published: June 2019

Insulin promotes glucose uptake by triggering the translocation of the glucose transporter GLUT4 from intracellular storage vesicles to the cell surface through exocytosis. Defects in GLUT4 exocytosis are a hallmark of insulin resistance (IR) and type 2 diabetes (T2D). To develop effective and safe treatments for IR and T2D, it is crucial to gain a comprehensive understanding of insulin-stimulated GLUT4 exocytosis at the molecular level. Recently, our group developed new genetic screen platforms to systematically dissect insulin-stimulated GLUT4 exocytosis, taking advantage of the revolutionary CRISPR-Cas9 genome editing system. Our genome-wide CRISPR screens recovered known GLUT4 exocytic regulators but most of the hits were not previously linked to the GLUT4 exocytic pathway. Here, I will focus on AAGAB, a negative regulator of GLUT4 translocation identified in the screens. We discovered that AAGAB plays a dual role in GLUT4 trafficking: 1) it promotes the endocytosis of GLUT4, and 2) it negatively regulates the SNARE-Munc18-mediated GLUT4 vesicle fusion. In addition, I will discuss OSBPL8 and OSBPL10, another two negative regulators identified in our screens. OSBPL8 and OSBPL10 are putative lipid transfer proteins operating between intracellular organelles. We propose that OSBPL8 and OSBPL10 regulate lipid dynamics required for insulin signaling, thereby influencing the activities of GLUT4 exocytic regulators. These genetic studies will facilitate the identification of new therapeutic targets for IR and T2D.

Disclosure

J. Shen: None.

Funding

National Institutes of Health
1915-P: Targeting Remission of Type 2 Diabetes Using a Digital Education and Behavior Change Program Improves Insulin Sensitivity and Liver Fat Content in Type 2 Diabetes
Authors: PESTA DOMINIK , ZAHARIA OANA P. , KUPRIYANOVA YULIYA , HWANG JONG-HEE , MARKGRAF DANIEL F. , TRENELL MICHAEL , SZENDROEDI JULIA , RODEN MICHAEL
Published: June 2019

Recent controlled trials have demonstrated that type 2 diabetes (T2D) remission is possible with significant weight loss in people with newly diagnosed T2D. However, current programs focus on face-to-face education and behavior change, which limit scalability and inclusivity. We examined the effect of a digital education and behavior change program targeting a very low-calorie diet on metabolic control, insulin sensitivity as well as liver fat and fibrosis.

Patients with diet/metformin controlled T2D (n=8, 50% male, 52±12 years, BMI 33±4 kg/m2) were remotely supported to undertake a very low-calorie (800 kcal/day) diet using digital education, behavior change and tracking with one-to-one tele-coaching for 12 weeks. Insulin sensitivity index was calculated using a mixed-meal tolerance test, liver fat content by 1H-magnetic resonance spectroscopy and liver fibrosis by elastography. After 12 weeks, participants had reduced their body weight by 9.0±3.6 kg and HbA1c (6.6±1.2% vs. 6.0±0.6%, p=0.05) and improved insulin sensitivity (319±61 vs. 355±49 ml.min-1.m-2, p<0.05). Liver fat content declined (9.8±8.2% to 3.3±3.1%, p<0.01) alongside with measures of liver fibrosis (7.0±3.1 to 4.4±1.3 kPa, p=0.05). Changes of liver fat were directly correlated with changes in body weight (r=0.80, p<0.05) as well as changes in fasting plasma glucose (r=0.85, p<0.01) and HbA1c (r=0.91, p<0.01). Within 12 weeks. 38% of participants achieved remission of T2D (HbA1c <6.5%) and 50% normalized liver fat (intrahepatic fat content <5.6%).

In conclusion, this feasibility study demonstrates that digital education, behavior change and tracking can reduce body weight which conveys significant improvements in metabolism and liver health. These preliminary data suggest that care teams should explore how digital education and behavior change can enhance existing care for people with newly diagnosed T2D with a view to achieving remission.

Disclosure

D. Pesta: None. O.P. Zaharia: None. Y. Kupriyanova: None. J. Hwang: None. D.F. Markgraf: Research Support; Self; Sanofi. M. Trenell: Other Relationship; Self; Changing Health Ltd. J. Szendroedi: None. M. Roden: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Poxel SA, Servier. Board Member; Self; Eli Lilly and Company. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi. Speaker's Bureau; Self; Novo Nordisk Inc.

Funding

EIT Health; German Ministry of Culture and Science of the State of North Rhine-Westphalia; German Federal Ministry of Health; German Federal Ministry of Education and Research
1922-P: Branched-Chain Ketoacid Dehydrogenase Kinase Inhibition Acutely Improves Glycemia and Alters the Response to Feeding in Mice
Authors: BOLLINGER ELIZA , SIDDALL CHARLES PARKER , GREIZER TIMOTHY , LIBERA JENNA L. , HARIRI GHAZAL , PASHOS EVANTHIA E. , SHIPSTONE ARUN , HADJIPANAYIS ANGELA , SUN ZHONGYUAN , XING GANG , CLASQUIN MICHELLE F. , ZHANG BEI B. , MILLER RUSSELL A. , ROTH FLACH RACHEL
Published: June 2019

Circulating increases in branched chain amino acid (BCAA) levels have long been associated with type 2 diabetes and metabolic syndrome. Emerging data also suggest that BCAA catabolism may play a role in heart failure progression. It is hypothesized that decreased catabolism, rather than increased consumption of BCAAs, is responsible for these correlations. Branched chain ketoacid (BCKA) dehydrogenase (BCKDH) kinase (BCKDK) is a negative regulator of BCAA catabolism through its inhibitory phosphorylation of the BCKDH E1a subunit. Using the BCKDK inhibitor molecule BT2, we demonstrate here a reduction of BCAA, BCKA and tissue p-BCKDH levels concomitant with improved glucose tolerance and reduced insulin levels in diet-induced obese mice after only one day of treatment. To investigate the mechanisms underlying this protection, we assessed plasma biomarkers and tissue gene expression after fasting and re-feeding in glucose intolerant high fat diet-fed animals. Remarkably, BT2 treated animals demonstrated reduced plasma glucose levels after re-feed, which was accompanied by dramatic reductions in plasma insulin levels, reduced activation of Akt in peripheral tissues, and a failure to suppress plasma free fatty acids and lipolytic machinery in adipose tissue. RNAseq was performed in liver to assess changes in gene expression profiles, and while over 400 genes were differentially regulated in vehicle treated re-fed mice compared with vehicle treated fasted mice, there were no differentially regulated genes in BT2 re-fed animals compared with BT2 fasted animals. These data suggest that activation of BCAA catabolism with the BCKDK inhibitor BT2 impairs the systemic response to feeding.

Disclosure

E. Bollinger: Employee; Self; Pfizer Inc. C.P. Siddall: Employee; Self; Pfizer Inc. Employee; Spouse/Partner; Vertex Pharmaceuticals. T. Greizer: None. J.L. Libera: None. G. Hariri: None. E.E. Pashos: Employee; Self; Pfizer Inc. A. Shipstone: None. A. Hadjipanayis: None. Z. Sun: None. G. Xing: None. M.F. Clasquin: Employee; Spouse/Partner; Merck & Co., Inc. Employee; Self; Pfizer Inc. B.B. Zhang: Employee; Spouse/Partner; Janssen Pharmaceuticals, Inc. Employee; Self; Pfizer Inc. Other Relationship; Self; Eli Lilly and Company. R.A. Miller: Employee; Self; Pfizer Inc. R. Roth Flach: Employee; Self; Pfizer Inc.
1924-P: Praliciguat, a Clinical-Stage sGC Stimulator, Improves Insulin Sensitivity, Lipid Tolerance, and Energy Utilization in a Mouse Diet-Induced Obesity Model Housed at Thermoneutrality
Authors: SCHWARTZKOPF CHAD , HADCOCK JOHN , ROUX JULIEN , LIU GUANG , SHEA COURTNEY , CURRIE MARK , MILNE G. TODD , MASFERRER JAIME , JONES JULI E.
Published: June 2019

Background: Praliciguat (IW-1973) is a soluble guanylate cyclase (sGC) stimulator under clinical investigation as a potential treatment for diabetic nephropathy. In animal models, praliciguat distributes broadly to tissues and elicits hemodynamic, anti-inflammatory, anti-fibrotic, and metabolic effects. Here, we assessed the metabolic effects of praliciguat in a mouse diet-induced obesity (DIO) model when housed at thermoneutrality.

Methods: Six week old male C57BL/6N mice were maintained on a low-fat diet (LFD, lean mice) or put on a 60% high-fat diet (HFD, obese mice). At 14 weeks of age, one group of obese mice was maintained on HFD (obese controls) while another group of obese mice was switched to HFD formulated with praliciguat to achieve a Cmax similar to a daily oral administration of 6 mg/kg (praliciguat treatment). A group of lean mice were also included (lean controls). Indirect calorimetry was performed on days 8, 9, 20, 21, 32 and 33. On day 35, an oral lipid tolerance test (LTT) was conducted. In a second study under the same dosing paradigm, overnight fasted blood and organs were collected on day 28.

Results: Compared to obese controls, energy expenditure was higher in praliciguat-treated mice throughout a 24h recording period as assessed by AUC (Day 8: +13%, Day 20: +9%, Day 21: +7%, Day 32: +5%, P<0.05) and LTT triglycerides were lower as assessed by AUC (-34%, P<0.01). In the second study, praliciguat-treated mice had lower fasting insulin (-28%, P<0.05), c-peptide (-20%, P<0.05), HOMA-IR (-26%, P<0.05) and triglycerides (-16%, P<0.01) compared to obese controls. Caloric intake, body weight and composition were unaffected by praliciguat treatment.

Conclusions: In a mouse DIO model housed at thermoneutrality, praliciguat increased energy utilization, improved triglycerides in response to an oral lipid challenge, improved insulin sensitivity, and lowered plasma triglycerides.

Disclosure

C. Schwartzkopf: Employee; Self; Ironwood Pharmaceuticals, Inc. J. Hadcock: Employee; Self; Ironwood Pharmaceuticals, Inc. J. Roux: None. G. Liu: None. C. Shea: Employee; Self; Ironwood Pharmaceuticals, Inc. M. Currie: Employee; Self; Ironwood Pharmaceuticals, Inc. G. Milne: Employee; Spouse/Partner; Catabasis Pharmaceuticals. Employee; Self; Ironwood Pharmaceuticals, Inc. Stock/Shareholder; Spouse/Partner; Catabasis Pharmaceuticals. Stock/Shareholder; Self; Ironwood Pharmaceuticals, Inc. J. Masferrer: Employee; Self; Ironwood Pharmaceuticals, Inc. J.E. Jones: None.
1927-P: Metabolomics of Normal Weight vs. Overweight Diabetics
Authors: FAROQI LIDA ,   IMPACT STUDY
Published: June 2019

The majority of people living with type 2 diabetes mellitus (T2DM) are overweight or obese due to the additional burden of applying insulin to properly control blood sugar levels. However, some people, especially Asian populations, develop T2DM with a low BMI (normal weight). This discrepancy may be resulted from differences in body fat and muscle mass for a given BMI. In order to investigate the influences of body weight on the onset of T2DM, we collected plasma samples from 9 overweight and 17 normal-weight diabetic patients as a pilot study, then applied untargeted metabolomics profiling on these plasma samples. By combining Hydrophilic Interaction (HILIC) and Reverse-Phase Liquid Chromatography (RPLC)-Mass Spectrometry, we comprehensively detected and relatively quantified 5,469 metabolites. We then employed various pattern recognition algorithms (principal component analysis, hierarchical clustering, Welsh t-test, etc.), clearly separated overweight and normal-weight diabetic groups based on their plasma metabolite profiling. Furthermore, we statistically identified 52 significantly different metabolites between these two groups, mainly involved in lipid metabolism. These results not only demonstrate that untargeted metabolomics is able to serve as an efficient diagnostic tool to distinguish overweight and normal-weight diabetic patients, but also provide meaningful insights into molecular mechanisms about tight relationships between diabetes and obesity, further facilitate biomarker discovery for early diagnosis and differentiation of normal/overweight diabetes.

Disclosure

L. Faroqi: None.
1934-P: Acute Hyperglycemia Increases Perfusion of Skeletal and Cardiac Muscle in Healthy Subjects
Authors: HORTON WILLIAM B. , JAHN LINDA , HARTLINE LEE , PATRIE JAMES T. , BARRETT EUGENE
Published: June 2019

Introduction: Over time, diabetes damages both the micro- and macrovasculature. Acutely, hyperglycemia causes oxidative stress and multiple biochemical abnormalities in endothelial cells. However, initial in vivo functional vascular responses to acute hyperglycemia (AH) are not well-characterized in humans or animals. Measurement of microvascular perfusion (MP) and endothelial function provides in vivo estimates of nutrient and hormone access to tissues and endothelial health, respectively.

Methods: Thirteen healthy adults (18-35 years, BMI 18-25 kg/m2) were fasted overnight. Octreotide was infused with basal insulin replacement and baseline measures of endothelial function by flow-mediated dilation (FMD) and MP (heart and skeletal muscle) by contrast-enhanced ultrasound were obtained after 90 minutes. Glucose was infused to maintain fasting euglycemia (EU) or raise glucose to ∼200 mg/dL (AH) for 4 hours with measures of FMD and MP repeated. Linear mixed model was used for statistical analysis, with Bonferroni correction for between-study comparisons.

Results: Compared to baseline, 4 hours of AH significantly increased skeletal muscle microvascular blood volume (MBV) (p=0.0002), microvascular flow velocity (p=0.047), and microvascular blood flow (p=0.025). MBV significantly increased in comparison to euglycemia (p=0.02). MBV also increased in cardiac muscle during AH (p=0.002). FMD trended towards a significant increase with AH (p=0.058). No changes were observed with EU.

Discussion: First, octreotide had no discernible effect on MP or FMD, which allowed testing the effect of AH while insulin remained at basal. Second, we provide the first evidence that heart and skeletal muscle respond to 4 hours of AH by increasing perfusion in healthy subjects. Whether this response is maintained with chronic hyperglycemia is unknown.

Disclosure

W.B. Horton: None. L. Jahn: None. L. Hartline: None. J.T. Patrie: None. E. Barrett: Research Support; Self; AstraZeneca.

Funding

National Institutes of Health (F32HL142304-01)
1942-P: Intermittent Fasting in Mice Improves Exercise Endurance through Enhancement of Mitochondrial Activating Genes in the Muscle
Authors: MIYASHITA KAZUTOSHI
Published: June 2019

Background: Intermittent fasting (IF) has been known to increase insulin sensitivity and lower body weight. However, the underlining mechanism for the beneficial effects has not been elucidated enough.

Design and Method: Mice were subjected to a severe IF regimen, in which they were fasted for 72 hours and refed ad libitum for subsequent 96 hours. This cycle was consecutively repeated 4 times to enforce the effects. After 4 cycles of the IF regimen, mice were fed a high-fat diet (HFD) ad-libitum for subsequent 4 weeks of observational period. At the 2 different time points: just after 4 cycles of the IF regimen and after subsequent 4 weeks of observational ad-libitum feeding period, the physical performance was examined.

Result: Glucose tolerance tests showed a significant decrease in the glucose level of IF group just after 4 cycles of the IF regimen. In PCR analysis, expressions of genes responsible for fat oxidation and mitochondrial activation (ACO et al.) were significantly increased in the muscle. The histone acetylation in the promoter region of these mitochondria activating genes was augmented and suggested the significance for enhancement of the expressions of these genes. In addition, we found that exercise endurance was improved in the IF group, without a change in the grip power. After subsequent 4 weeks of observational feeding period, the BW in IF group was significantly lower than ad-libitum feeding group. At the time-point, the exercise endurance, glucose tolerance, histone acetylation and expression of genes maintained the similar changes observed just after 4 cycles of the IF regimen.

Conclusion: The 72 hours fasting IF regimen augmented histone acetylation in the promoter regions of mitochondrial activating genes in the muscle and adipose tissues that would accounts for the prolonged enhancement of the expression. We found that not only glucose tolerance but also exercise endurance was improved through the IF procedure.

Disclosure

K. Miyashita: Research Support; Self; Johnson & Johnson, Merck & Co., Inc., Novo Nordisk Inc.
1943-P: The Sodium-Glucose Cotransporter 2 Inhibitor Luseogliflozin Can Suppress Muscle Atrophy in Db/Db Mice by Suppressing the Expression of Foxo1
Authors: BAMBA RYO , KIMURA TOMONORI , OKAMURA TAKURO , HASHIMOTO YOSHITAKA , OSAKA TAKAFUMI , SENMARU TAKAFUMI , FUKUI MICHIAKI
Published: June 2019

Here we investigated the effect of the sodium glucose cotransporter-2 inhibitor (SGLT2i) luseogliflozin on skeletal muscle. Eight-week-old mice were fed a standard diet or the standard diet with added luseogliflozin for 8 weeks. The mice were divided into the following four genotype/dietary groups: Db/m mice without (w/o) SGLT2i, Db/m mice with SGLT2i, Db/Db w/o SGLT2i, and Db/Db with SGLT2i. After the 8-week dietary treatment, we investigated the changes in body weight, glucose tolerance, and muscle in each group. Among the mice with and w/o SGLT2i, the ratio of soleus and plantaris muscle to body weight in the Db/Db mice was significantly lower than that in the Db/m mice. The cross-sectional area of soleus muscle in the Db/Db mice w/o SGLT2i was significantly higher than that in the Db/Db mice with SGLT2i. The expression of foxo1 in soleus muscle of the Db/Db mice was significantly higher than that of the Db/m mice, and the foxo1 expression of the Db/Db mice with SGLT2i was significantly lower than that of the mice w/o SGLT2i. The fluorescence intensity of foxo1 in the Db/Db mice fed SGLT2i was significantly lower than that in the Db/Db mice w/o SGLT2i.The administration of luseogliflozin resulted in the suppression of both the increased foxo1 expression and the reduced muscle cross-sectional area in the soleus muscle of Db/Db mice.

Disclosure

R. Bamba: None. T. Kimura: None. T. Okamura: None. Y. Hashimoto: Research Support; Self; Asahi Kasei Pharma. T. Osaka: None. T. Senmaru: None. M. Fukui: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Eli Lilly and Company, Johnson & Johnson, Kyowa Hakko Kirin Co., Ltd., Merck & Co., Inc., Mitsubishi Tanabe Pharma Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi, Sanwa Kagaku Kenkyusho, Takeda Pharmaceutical Company Limited. Speaker's Bureau; Self; AstraZeneca, Eli Lilly and Company, Kowa Pharmaceutical Europe Co. Ltd., Kyowa Hakko Kirin Co., Ltd., Merck & Co., Inc., Mitsubishi Tanabe Pharma Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi, Sanwa Kagaku Kenkyusho, Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited.
1944-P: Muscle a-Parvin Is Not Necessary for Integrin-Linked Kinase Related Insulin Resistance in Diet-Induced Obese Mice
Authors: CAPPEL DAVID A. , POZZI AMBRA , ZENT ROY , WASSERMAN DAVID
Published: June 2019

High fat diet (HFD) causes obesity and expansion of extracellular matrix (ECM) components and causes insulin resistance. Integrins provide for adhesion between cells and the ECM and signal cells information about the local environment. A major component of signaling through the integrin β1 subunit is the integrin-linked kinase (ILK). Previous studies demonstrated mice that do not express ILK in skeletal muscle (SkM) are resistant to HFD-induced muscle insulin resistance. ILK has no catalytic activity but is the scaffold for signaling molecules, including α-parvin which regulates Rho-GTPases which are involved in actin cytoskeleton and vesicular trafficking. To test the hypothesis that ILK-related insulin resistance is mediated through α-parvin in SkM, we generated a mouse model which lacks α-parvin in SkM (mParKO) using a cre-lox system under control of HSA promoter. mParKO mice and non-transgenic littermates (WT) were fed either chow or HFD for 12 weeks after which insulin sensitivity was assessed in vivo using the hyperinsulinemic-euglycemic clamp combined with 3-[3H]glucose and 2-[14C]deoxyglucose tracers to measure glucose fluxes. Body weight was not different between mParKO and WT mice on either diet, nor was fasting blood glucose. In contrast to previous clamp studies of HFD-fed muscle ILK knockout mice, clamp glucose infusion rate, tracer-determined glucose rates of appearance and disappearance, and an index of SkM glucose uptake were not different between mParKO and WT mice on either diet. These results suggest that the α-parvin unit does not mediate the profound ILK-related susceptibility to insulin resistance in HFD fed mice.

Disclosure

D.A. Cappel: None. A. Pozzi: None. R. Zent: None. D. Wasserman: None.

Funding

National Institutes of Health (DK59637, DK50277, DK54902); American Heart Association
1950-P: Effects of Dietary Composition on Intestinal Glucocorticoid Receptor Signaling
Authors: RADLINGER BERNHARD , FOLIE SABRINA , DOBNER JOCHEN , ADOLPH TIMON-ERIK , RESS CLAUDIA , HORNSTEINER FLORIAN , BOENTGES SARAH , SALZMANN KARIN , TILG HERBERT , KASER SUSANNE
Published: June 2019

Enhanced glucose transport capacity of the intestinal epithelium, specifically enterocytes, is known to significantly contribute to systemic glucocorticoid (GC) - induced hyperglycemia. This effect is mainly dependent on overexpression of sodium glucose transporter 1 (Sglt1) and its enhanced activation by serum and glucocorticoid-regulated kinase 1 (Sgk1). In this work we aimed to investigate whether dietary intake affects intestinal glucocorticoid receptor (Nr3c1) signaling. In male C57/BL6 mice short term feeding of a Western diet (WD) resulted in increased glucocorticoid receptor expression in the ileum which was accompanied by an upregulation of the effector genes Sgk1 and sodium-hydrogen exchanger 3 (Nhe3) and liver receptor homologe 1 (Lrh1). The latter is known as the key regulator of intestinal corticosterone synthesis. Histological analysis showed that glucocorticoid receptor expression was mainly located in enterocytes. Intestinal Sgk1 expression remained significantly, about 2-fold, increased in mice fed a Western diet for 10 weeks, while Nr3c1, Lrh1 and Nhe3 expression levels were not significantly altered after long-term feeding. Further regulatory mechanisms were investigated in immortalized murine small intestine enterocyte cell line Mode-K: Stimulation with palmitic acid resulted in a significant upregulation of Sgk1 expression, while no effect was seen when exposed to oleic acid, arachidonic acid or docosahexanoic acid. In contrast to Sgk1, Nr3c1 expression was downregulated in Mode-K cells exposed to palmitic acid.

In conclusion, our data suggest that dietary fat intake might contribute to GC induced hyperglycemia by modulating intestinal Sgk1 expression in a glucocorticoid receptor-independent way.

Disclosure

B. Radlinger: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp. S. Folie: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp. J. Dobner: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp. T. Adolph: None. C. Ress: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp. F. Hornsteiner: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp. S. Boentges: Research Support; Self; Boehringer Ingelheim International GmbH, Merck & Co., Inc. K. Salzmann: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp. H. Tilg: None. S. Kaser: Research Support; Self; Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp.

Funding

Austrian Federal Ministry of Science, Research and Economy; National Foundation for Research, Technology and Development; Merck Sharp & Dohme Austria; Boehringer Ingelheim RCV