bluuebunny/crossref_metadata_2025 huggingface dataset:
Mitochondrial Function in High-Fat Fed and Exercise-Trained Mice Unable to Induce Autophagy
Authors: EHRLICHER SARAH E. , STIERWALT HARRISON D. , NEWSOM SEAN A. , ROBINSON MATTHEW M.
Published: June 2018

Impaired mitochondrial lipid respiration is implicated in obesity-induced insulin resistance but can be enhanced with exercise training. Autophagy is involved in remodeling the mitochondrial proteome and may be required for mitochondrial and whole-body adaptations in lipid respiration. We used a diet-induced obesity model of C57BL/6J mice and investigated the requirement of autophagy for mitochondrial adaptations to 8 weeks of treadmill exercise training using transgenic mice that cannot induce autophagy with exercise (BCL) compared to wild type controls (WT, n=3-5). High fat (HF) fed mice had lower glucose tolerance at 4 weeks compared to low fat (LF, ∼35%, P<0.01), regardless of genotype. After 8 weeks of exercise, LF-BCL mice had similar glucose tolerance to LF-WT mice. However, the HF-BCL mice had lower glucose tolerance compared to HF-WT mice (∼19%, P<0.01) suggesting a blunted improvement in glucose tolerance following training in the absence of autophagy. At week 12, mitochondria were isolated from quadriceps for respirometry. Oxidative phosphorylation capacity was normalized to electron transport system capacity. Lipid supported respiration (octanoyl-carnitine) was higher in the HF mice compared to LF (∼17%, P<0.01) and the lack of autophagy activation had no effect. Glutamate-malate-succinate supported respiration was lower in BCL mice compared to WT (∼11%, P<0.05), but only on a LF diet suggesting the requirement of autophagy for improved mitochondrial function with exercise may vary between substrates. Whole body respiratory exchange ratio was measured during 24 hours of fasting and all mice shifted to lipid oxidation with no difference between diet or genotype (P = 0.11). Collectively, skeletal muscle mitochondrial lipid oxidation was greater with HF, despite glucose intolerance. The activation of autophagy with exercise was not required for mitochondrial lipid adaptations or whole body metabolic flexibility, but may be required for non-lipid substrates.

Disclosure

S.E. Ehrlicher: None. H.D. Stierwalt: None. S.A. Newsom: None. M.M. Robinson: None.
Mono- and Co-Activation of the GIP and GLP-1 Receptors Inhibits Bone Resorption
Authors: BERGMANN NATASHA C. , LUND ASGER B. , HEIMBÜRGER SEBASTIAN M. , JESSEN LENE , CHRISTENSEN MIKKEL B. , VILSBØLL TINA , KNOP FILIP K.
Published: June 2018

We investigated the effect of mono- and co-administration of the incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) on bone resorption and formation. Seventeen overweight/obese men without diabetes were included in a randomized, double-blinded, placebo-controlled, cross-over study and subjected to a 50g oral glucose tolerance test (OGTT) and four isoglycemic intravenous glucose infusions (IIGI), mimicking the glucose excursions from the OGTT, with concomitant infusions of GIP (4 pmol/kg/min), GLP-1 (1 pmol/kg/min), GIP+GLP-1 (4 and 1 pmol/kg/min, respectively) or placebo. Bone resorption and formation were assessed by measurements of carboxy-terminal type 1 collagen crosslinks (CTX), and procollagen type 1 N-terminal propeptide (P1NP), respectively. Suppression of CTX is shown in Fig. 1. During OGTT the baseline subtracted area under the curve (bsAUC) was significantly lower than during IIGI+saline infusion (-9,117±566 (mean±SEM) vs. -4,333±583 %×min, P<0.0001). Infusion of GLP-1 and GIP significantly potentiated CTX suppression to -10,751±799 %×min and 13,714±440 %×min, respectively. Co-administration of GIP+GLP-1 suppressed CTX to -15,466±268 %×min. P1NP levels were unaffected. Our data suggest that GLP-1, like GIP, may be involved in bone homeostasis and that co-administration of GIP+GLP-1 inhibits bone resorption more than each hormone alone.

Disclosure

N.C. Bergmann: Employee; Self; Zealand Pharma A/S. A.B. Lund: Other Relationship; Self; Novo Nordisk A/S. S.M. Heimbürger: Research Support; Self; Novo Nordisk A/S. L. Jessen: Employee; Self; Zealand Pharma A/S. Stock/Shareholder; Self; Zealand Pharma A/S, Novo Nordisk A/S. M.B. Christensen: None. T. Vilsbøll: Advisory Panel; Self; Amgen Inc., Boehringer Ingelheim GmbH, Eli Lilly and Company, AstraZeneca, MSD K.K., Sanofi, Novo Nordisk A/S. Consultant; Self; Amgen Inc., Boehringer Ingelheim GmbH, Eli Lilly and Company, AstraZeneca, MSD K.K., Sanofi, Novo Nordisk A/S. Speaker's Bureau; Self; Amgen Inc., Boehringer Ingelheim GmbH, Eli Lilly and Company, AstraZeneca, MSD K.K., Sanofi, Novo Nordisk A/S. Research Support; Self; Eli Lilly and Company, Novo Nordisk A/S. F.K. Knop: Advisory Panel; Self; AstraZeneca. Consultant; Self; AstraZeneca. Research Support; Self; AstraZeneca. Speaker's Bureau; Self; AstraZeneca. Advisory Panel; Self; Eli Lilly and Company. Speaker's Bureau; Self; Eli Lilly and Company. Advisory Panel; Self; Amgen Inc.. Speaker's Bureau; Self; Merck Sharp & Dohme Corp.. Advisory Panel; Self; Novo Nordisk A/S. Consultant; Self; Novo Nordisk A/S. Speaker's Bureau; Self; Novo Nordisk A/S. Consultant; Self; Amgen Inc.. Advisory Panel; Self; MedImmune, Sanofi. Consultant; Self; Sanofi. Research Support; Self; Sanofi. Speaker's Bureau; Self; Sanofi.
Hepatic Insulin Clearance Is Increased in the Patients with Poorly Controlled Type 2 Diabetes
Authors: OKURA TSUYOSHI , ANNO MARI , ITO YUICHI
Published: June 2018

Aims: The hepatic insulin clearance (HIC) is an important pathophysiology of type 2 diabetes mellitus (T2DM). Some studies reported that HIC was decreased in the patients with T2DM. However, a basic research reported that hyperglycemia enhances HIC, and it is not well-known whether poorly controlled diabetes increases HIC in the patients with T2DM. We investigated whether HIC was increased in the patients with poorly controlled diabetes, and we also evaluated whether HIC associated with insulin resistance.

Methods: We performed meal tolerance test and hyperinsulinemic-euglycemic clamp for 23 patients with type 2 diabetes (mean HbA1c 7.5%). We calculated the postprandial C-peptide AUC/insulin AUC ratio as HIC.

Result: The HIC significantly correlated with HbA1c (R=0.60, P<0.005). In the patients with high HIC above the median, the mean HbA1c was significantly higher than that of low HIC below the median (7.95% vs. 6.99%, P<0.01). The HIC was not correlated with insulin resistance as evaluated by the glucose clamp method (R=0.29, P=0.16).

Conclusion: These results suggested that hepatic insulin clearance was increased in the patients with poorly controlled T2DM, especially in the patients with HbA1c>8%.

Disclosure

T. Okura: None. M. Anno: None. Y. Ito: None.
Lower Risk of Dementia in Elderly Patients Initiated on Dipeptidyl Peptidase-4 Inhibitor vs. Sulfonylurea—A Population-Based Cohort Study
Authors: HAN SEUNG JIN , JEON JA YOUNG , LEE KWANWOO , KIM TAE HO , MOON HYUN UK
Published: June 2018

Background: Type 2 diabetes is related with an increased risk of dementia. Dipeptidyl peptidase-4 inhibitors (DPP-4i) have shown promising results for its use in dementia in preclinical studies. Therefore, we investigated the risk of developing dementia in elderly patients initiated on DPP-4i vs. sulfonylurea (SU).

Methods: A population-based cohort study using claims database named the Korean National Health Insurance Service Senior cohort (ver. 3.0, 1 January 2002 to 31 December 2015) was performed. DPP-4i-treated patients and SU-treated patients were matched by 1:1 propensity score matching which was calculated with 49 confounding variables. Kaplan-Meier curves and Cox proportional hazards regression analysis were performed to estimate the risk of dementia among patients who were prescribed DPP-4i compared with patients who were prescribed SU.

Results: In total, 7,561 patients on each group were paired using propensity score matching. The risk of dementia was lower in the DPP-4i group compared to the DPP-4i group (hazard ratio [HR] 0.66; 95% confidence interval [CI] 0.56-0.79; P<0.001). Also, HR of Alzheimer’s dementia, and vascular dementia were lower in DPP-4i-treated patients compared with SU-treated patients (HR 0.71; 95% CI 0.57-0.89); P=0.002 for Alzheimer’s dementia, HR 0.50; 95% CI 0.30-0.85); P=0.01 for vascular dementia).

Conclusions: Our findings suggest that DPP-4i use decrease the risk of dementia compared with SU.

Disclosure

S. Han: None. J. Jeon: None. K. Lee: None. T. Kim: None. H. Moon: None.
Deletion of OGlcNAc Transferase in Pancreatic Progenitors Causes Partial Pancreas Agenesis and Diabetes Independent of p53 in Mice
Authors: ALEJANDRO EMILYN , BAUMANN DANIEL , JO SEOKWON , AKHAPHONG BRIAN , LOCKRIDGE AMBER D. , MOHAN RAMKUMAR
Published: June 2018

Deletion of the only nutrient-sensor enzyme, O-GlcNAc Transferase (OGT), catalyzing O-GlcNAc addition onto target proteins in pancreatic β-cells induces diabetes through β-cell failure. We hypothesized that OGT, expressed at a very high level in β-cells, has key developmental regulatory properties. In this study, we tested the hypothesis that OGT is required for pancreas and β-cell development by deleting OGT in pancreatic (Pdx1Cre, OGTKOPanc) or endocrine (Ngn3Cre, OGTKOEndo) progenitors. We show that mice lacking OGT in pancreatic progenitors (Pdx1+, OGTKOPanc) show near pancreas agenesis and loss of β-cell mass at birth. Few mice that grow into adulthood develop overt diabetes due to functional β-cell mass failure. OGT ablation in endocrine progenitors (Ngn3+, OGTKOEndo) resulted in normal pancreas mass but reduced β-cell mass and diabetes early in life, strongly revealing the temporal and importance of OGT for β-cell development. We observed loss of Pdx1 staining at birth and on embryonic (e) day 14.5 in OGTKOPanc. Transcriptome analysis of e14.5 progenitor single cells show distinct cell clustering populations in OGTKOPanc compared to littermate controls. Upstream regulator analysis of altered genes show enhanced activation of p53, also known as TP53 or tumor protein. Next we identified p53 to be O-GlcNAc-modified in β-cells. Concomitant deletion of p53 in the OGTKOPanc background (p53,OGTKOPanc) does not rescue pancreas agenesis in OGTKOPanc newborn, and adult p53,OGTKOPanc mice develop severe hyperglycemia and diabetes. These data highlight the novel role O-GlcNAc-modifications in pancreas and β-cell development and provide a new model to characterize the developmental defects associated with pancreatic agenesis.

Disclosure

E. Alejandro: None. D. Baumann: None. S. Jo: None. B. Akhaphong: None. A.D. Lockridge: None. R. Mohan: None.
Regulation of Lipid Regulator ANGPTL8 Gene Transcript by Insulin Is Mediated by C/EBPß, and ANGPTL8 Deficiency Improves Glucose Homeostasis in Mice
Authors: ZHANG LU , ABDUL-GHANI MUHAMMAD , DEFRONZO RALPH A. , NORTON LUKE
Published: June 2018

Angiopoietin-like protein 8 (ANGPTL8) is a novel circulating protein synthesized by the liver and adipose tissue and plays a key role in regulating the plasma lipid profiles. Its expression is regulated acutely by fasting and refeeding in humans and animals. However, the molecular and physiological mechanisms, which regulate its expression, remain largely unknown. In this study, we investigated the regulation of ANPTL8 by insulin and glucose in multiple cellular and animal systems. Interestingly, insulin alone rapidly and dose-dependently increased ANGPTL8 gene expression in both cultured hepatocytes and mature adipocytes, while high glucose combined with insulin presented further stimulation on ANGPTL8 only in adipocytes. In mice, 2-hour euglycemic-hyperinsulinemic and hyperglycemic-hyperinsulinemic clamps markedly stimulated ANGPTL8 mRNA levels in liver to a similar extent. However, only combined hyperglycemia-/hyperinsulinemia caused a significant elevation of ANGPTL8 gene transcript in mouse adipose tissue. These findings imply that distinct pathways are responsible for the effects of insulin and glucose on ANGPTL8 expression in liver and adipose tissue. By cloning the ANGPTL8 gene promoter fragment into a luciferase reporter vector, we discovered that insulin-induced ANGPTL8 gene transcript was mediated by the CCAAT/enhancer-binding protein beta (C/EBPβ) transcription factor, which is likely through Janus Kinase 2 (JAK2) pathway. We also observed that AMPK activation had an inhibitory effect on insulin-induced ANGPTL8 gene expression; this could explain, in part, the low ANGPTL8 levels during fasting. Finally, in mice ANGPTL8 knockout did not alter body weight or food intake compared to wild type littermates, but significantly improved glucose tolerance in diet induced obese mice. This beneficial effect on glucose tolerance most likely is explained by the reduction in serum FFA and triglyceride levels.

Disclosure

L. Zhang: None. M. Abdul-Ghani: None. R.A. DeFronzo: Speaker's Bureau; Self; AstraZeneca, Novo Nordisk Inc.. Advisory Panel; Self; AstraZeneca, Novo Nordisk Inc., Janssen Pharmaceuticals, Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Elcelyx Therapeutics, Inc., Intarcia Therapeutics, Inc.. Research Support; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Pharmaceuticals, Inc., Takeda Pharmaceuticals U.S.A., Inc.. L. Norton: None.
Effects of Simple Carbohydrates on GLP-1 Responses in Gastric Bypass Patients and Matched Controls
Authors: MARTINUSSEN CHRISTOFFER , BOJSEN-MOLLER KIRSTINE N. , DIRKSEN CARSTEN , SVANE MARIA S. , HOLST JENS J. , MADSBAD STEN
Published: June 2018

Exaggerated postprandial secretion of GLP-1 seems to contribute importantly to the metabolic effects of Roux-en-Y gastric bypass (RYGB). The surgery accelerates nutrient entry to the distal intestine, which is believed to augment GLP-1 secretion. In un-operated individuals, slowly digested carbohydrates are also absorbed in the distal intestine and may accordingly produce greater GLP-1 responses than rapidly absorbed ones.

Our aim was to investigate GLP-1 secretion in response to isomolar (0.1385 mol) oral loads of glucose (glu) and fructose (fru), either given as separate monosaccharides (25 g glu and 25 g fru) or as disaccharides in the form of 47.5 g sucrose (suc) +/- alpha glucosidase inhibition (by acarbose (aca)) or 47.5 g isomaltulose (iso), which is slower digested than suc, on 4 separate days in 10 RYGB patients and 10 controls (CON) matched on BMI, age and sex.

Glu/fru and suc induced similar glucose excursions and responses of C-peptide and GLP-1, the latter being 3-fold greater in RYGB compared to CON. Digestion of iso was slow in comparison with suc in both groups. Mean GLP-1 response (positive iAUC) to iso was ∼50% and ∼160% greater than to suc in RYGB (p=0.11) and CON (p=0.01), respectively. Aca effectively slowed suc digestion in both groups, resulting in lower peak concentrations of glucose (RYGB: 9.3 vs. 6.7 mmol/l, CON: 7.7 vs. 6.6 mmol/l) and C-peptide (RYGB: 2762 vs. 1341 pmol/l, CON: 2367 vs. 1520 pmol/l) compared with suc alone (p<0.01). Aca diminished mean suc stimulated GLP-1 secretion by ∼50% in RYGB (p<0.01), but this effect was absent in CON.

The slowly absorbed disaccharide isomaltulose elicits a greater GLP-1 response than sucrose. The same effect is not evident when digestion of sucrose is inhibited by acarbose, which in RYGB diminishes sucrose induced GLP-1 secretion. Understanding the digestive processes involved in carbohydrate induced GLP-1 release after gastric bypass is an important step towards mimicking the effects of the surgery by pharmacological means.

Disclosure

C. Martinussen: None. K.N. Bojsen-Moller: None. C. Dirksen: None. M.S. Svane: None. J.J. Holst: Advisory Panel; Self; Novo Nordisk A/S. Board Member; Self; Zealand Pharma A/S. Speaker's Bureau; Self; Merck Sharp & Dohme Corp., AstraZeneca. Research Support; Self; Danish Diabetes Academy, Novo Nordisk Foundation. Other Relationship; Self; Antag Therapeutics. Other Relationship; Spouse/Partner; Antag Therapeutics. S. Madsbad: Advisory Panel; Self; AstraZeneca. Speaker's Bureau; Self; AstraZeneca. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc.. Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc.. Speaker's Bureau; Self; Eli Lilly and Company, Eli Lilly and Company. Advisory Panel; Self; Merck Sharp & Dohme Corp., Sanofi-Aventis, Novo Nordisk A/S. Speaker's Bureau; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S.
Forced Expressed GLP-1R Attenuates Prostate Cancer Growth
Authors: SHIGEOKA TORU , NOMIYAMA TAKASHI , KAWANAMI TAKAKO , HAMAGUCHI YURIKO , TANAKA TOMOKO , YANASE TOSHIHIKO
Published: June 2018

Incretin therapy is one of the most popular treatment for type 2 diabetes. GLP-1R agonist has received much attention, because of its tissue protective effects beyond glycemic control. We have previously reported anti-prostate and anti-breast cancer effect of GLP-1R agonist Exendin-4(Ex-4) (Diabetes 2014, PLOS ONE 2015, Endocrinology 2017). In our previous report, the attenuation of cancer cell proliferation was depending on GLP-1R expression, and Ex-4 did not attenuate cell proliferation in ALVA-41, prostate cancer cell line without GLP-1R expression. Then, we next examined anti-prostate cancer effect in ALVA-41 forced expressed GLP-1R using lentivirus vector (ALVA-41-GLP-1R). We confirmed abundant GLP-1R expression in ALVA-41-GLP-1R using RT-PCR and immunohistochemistry. Ex-4 significantly decreased the proliferation of ALVA-41-GLP-1R in a dose dependent manner, but not in ALVA-41 transfected with control vector (ALVA-41-control), suggesting that forced expressed GLP-1R was intact. In the growth curve, cell number of ALVA-41-GLP-1R was significantly decreased compared with ALVA-41-control. BrdU assay revealed that cell proliferation was attenuated in ALVA-41-GLP-1R, and S Phase entry was inhibited in cell cycle distribution. Further, p27(CDKN1B) expression was increased in ALVA-41-GLP-1R compared with ALVA-41-control in Western Blotting analysis. These data suggest that forced expressed GLP-1R attenuated prostate cancer cell proliferation via inhibiting cell cycle progression.

Disclosure

T. Shigeoka: None. T. Nomiyama: Research Support; Self; MSD K.K., Sanofi-Aventis, Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceuticals, Japan. T. Kawanami: None. Y. Hamaguchi: None. T. Tanaka: None. T. Yanase: Research Support; Self; MSD K.K., Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceuticals, Japan.
The Role of Visceral Adipocytes in Adrenergically Stimulated Adiponectin Secretion
Authors: MUSOVIC SALIHA , NGUYEN SANDRA , OLOFSSON CHARLOTTA S.
Published: June 2018

Adiponectin is a white adipocyte hormone with insulin sensitizing properties. Correlations have been observed between adiponectin and different adipose tissue depots where serum adiponectin levels exhibit a negative relationship to visceral fat mass (VAT). Individuals with increased VAT have a greater risk for developing type 2 diabetes. We have previously shown that catecholamines stimulate acute adiponectin release from subcutaneous adipocytes via adrenergic beta-3-receptors (β3ARs) and Epac1. We further showed that this adrenergic stimulation was abolished in obesity/diabetes due to reduced levels of both β3ARs and Epac1. Here we aim to study the adrenergically stimulated adiponectin secretion in visceral adipocytes in health and in metabolic disease. Results obtained by ELISA revealed that epinephrine (EPI) and CL316243 (CL;β3AR-agonist) stimulated adiponectin release to a similar degree (∼2 fold) in primary visceral mouse adipocytes during 30 minutes incubation. The stimulatory effect of EPI was inhibited by Epac antagonist ESI-09 (P<0.05). Studies using visceral adipocytes isolated from obese/diabetic mice demonstrated that adiponectin release could still be stimulated with EPI and CL (∼1.5 fold) although at a significantly lower magnitude compared to control mice (P<0.001). RT-qPCR results showed a high expression of β3AR as well as Epac1 in adipocytes isolated from control mice. In adipocytes isolated from obese/diabetic mice there was no change in expression of β3AR but an increase of Epac1 (P<0.001). Our results propose that acute adiponectin release in visceral adipocytes is regulated through activation of β3AR and Epac1. It is an interesting finding that VAT adipocytes which are connected to several metabolically active organs largely maintain adrenergically stimulated adiponectin release in obesity/diabetes.

Disclosure

S. Musovic: None. S. Nguyen: None. C.S. Olofsson: None.
Insulin-Like Growth-Factor Axis Collectively Identifies Pre–Type 1 Diabetes
Authors: SHAPIRO MELANIE , WASSERFALL CLIVE , SCHULTZ ANDREW R. , MCGRAIL SEAN M. , HALLER MICHAEL J. , SCHATZ DESMOND , ATKINSON MARK A. , BRUSKO TODD M.
Published: June 2018

Autoantibodies (AAb) against beta cell antigens are useful biomarkers for type 1 diabetes (T1D) prediction; however, staging of pre-T1D requires lengthy and invasive glucose tolerance testing. A need exists for additional serum biomarkers reflective of metabolic and immunologic dysregulation to test concurrently with AAb. The insulin-like growth factor (IGF) family, consisting of IGF1 and IGF2, has been shown to promote insulin action and possess immunoregulatory properties. Their activity is regulated by at least seven IGF binding proteins (IGFBPs). We hypothesized that IGF1 and IGF2 levels or bioavailability are reduced during T1D development. Total serum IGF1, IGF2, and IGFBP1-7 levels were measured in an age-matched (12.0 ± 3.7 years), cross-sectional cohort of AAb negative (AAb-) controls (CTRL) (n=74); AAb- first-degree relatives (AAb- FDR, n=55); FDR positive (AAb+ FDR) for one (n=23) or multiple (n=26) of GAD65, IA-2, or ZnT8 AAbs; new onset (NO) T1D patients (n=63, duration 1.0 ± 0.7 months); and established T1D patients (n=68, duration 4.4 ± 3.7 years). IGF1 and IGF2 levels were significantly lower in AAb+ FDR compared to AAb- FDR (204 ± 117 vs. 346 ± 160 ng/mL, p<0.001; 701 ± 496 vs. 1054 ± 721 ng/mL, p<0.01). Lower IGF1 levels were noted in single AAb+ FDR (183 ± 112 ng/mL, p<0.0001), while lower IGF2 levels remained significant in multiple AAb+ FDR only (686 ± 535 ng/mL, p<0.05). IGFBP1 levels were increased in AAb+ FDR compared to AAb- FDR (3.6 ± 3.2 vs. 1.8 ± 1.7 ng/mL, p<0.01), and this was apparent in single AAb+ FDR (4.5 ± 4.0 ng/mL, p<0.001). IGFBP7 levels were significantly lower in multiple AAb+ FDR as compared to AAb- FDR (79 ± 21 vs. 95 ± 22 ng/mL, p<0.05). Additionally, we established a multiple logistic regression model evaluating age, IGFs, and IGFBPs (IGFBP1, -3, and -7), which could effectively discriminate AAb+ FDR from CTRL (AUC = 0.84). These data suggest a dysregulated IGF axis is present prior to symptomatic T1D and may provide new biomarkers to improve disease prediction.

Disclosure

M. Shapiro: None. C. Wasserfall: None. A.R. Schultz: None. S.M. McGrail: None. M.J. Haller: None. D. Schatz: None. M.A. Atkinson: Other Relationship; Self; Patent Issued. T.M. Brusko: Stock/Shareholder; Self; OneVax, LLC. Advisory Panel; Self; Caladrius Biosciences, Inc.. Consultant; Self; Merck & Co., Inc., Sanofi-Aventis.
Cross-Sectional Associations between Drinking Bottled Water and Prediabetes/Diabetes
Authors: JOSHIPURA KAUMUDI J. , TRAK MARIA A. , MUÑOZ-TORRES FRANCISCO J.
Published: June 2018

Dietary recommendations for healthy adults include adequate hydration (males: 125 ounces/day, females: 91 ounces/day) from drinking water, other beverages, and water contained in food. Bottled water sales in U.S. exceeded 11.7 billion gallons in 2015 (36.5 gallons per capita). Low water intake, as well as some chemicals in water sources, are associated with hyperglycemia. However, the association between drinking water sources and hyperglycemia/diabetes has not been studied. In a cohort of overweight/obese Puerto Ricans, we evaluated the association between water sources (bottled and filtered compared to tap water) and prediabetes/diabetes. In 2014-2016, 1023 participants reported their primary source of water as bottled (52%), filtered (25%) and tap (23%); diabetes status was assessed using ADA criteria for fasting and 2-hour post load glucose, and HbA1c. Logistic regression models controlled for age, gender, waist circumference, smoking, alcohol intake, physical activity, and hypertension. Bottled water consumers (compared to tap water) had higher prediabetes (OR=1.43; 95% CI: 1.02, 2.01), and diabetes (OR=1.93; 95% CI: 1.08, 3.46). Further controlling for amount of water, sugar sweetened beverages, or canned food and drinks did not change the associations, but controlling for education or income strengthened the association with diabetes. Results were stronger when restricted to San Juan city residents (43%): prediabetes (OR=1.68; 95% CI: 1.01, 2.79), and diabetes (OR=4.05; 95% CI: 1.48, 11.1). Filtered water users had somewhat higher prediabetes/diabetes (OR=1.38; 95% CI: 0.93, 2.04) compared to tap water users. Results suggest that drinking bottled water may be associated with higher prevalence of diabetes compared to tap water, potentially mediated by endocrine disruptors in plastic bottles. We cannot determine time sequence or causality, given the cross-sectional analyses. The impact of water sources on diabetes risk needs to be further evaluated longitudinally.

Disclosure

K.J. Joshipura: None. M.A. Trak: None. F.J. Muñoz-Torres: None.
Exendin-4, but Not Oleoylethanolamide, Promotes Host-Mediated Alterations to the Gut Microbiome during Weight Loss
Authors: BROWN JACOB , SHARMA VANDANA , TRAN DAVID , PETERSON SCOTT N. , AYALA JULIO , BROWN JACOB
Published: June 2018

Gut-derived molecules like glucagon-like peptide-1 (GLP-1) and oleoylethanolamide (OEA) are prime targets for developing more effective obesity therapeutics due to their weight loss action. Hypophagia is commonly cited as the mechanism by which OEA and Exendin-4 (Ex4), a GLP-1 receptor agonist, reduce body weight. However, we recently showed in diet-induced obese (DIO) mice that the hypophagic action of Ex4 and OEA is transient and that weight loss continues even as food intake returns to pre-treatment levels. This was associated with increased energy expenditure (EE) in Ex4-, but not OEA-treated, mice. Given the emerging role for the gut microbiome as a regulator of energy balance, we hypothesized that the long-term, food intake-independent weight loss elicited by Ex4 is associated with alterations in the gut microbiome. In vivo and in vitro experiments coupled with 16S analysis were used to determine the effect of chronic Ex4 and OEA on the gut microbiome during weight loss. Male, DIO C57Bl/6J mice (60% high-fat diet for 15 weeks) were dosed 2x daily with Vehicle, OEA (5mg/kg) or Ex4 (100µg/kg) for 7 days. Fecal pellets were collected pre- and post-treatment for 16S analysis. Ex4 caused a change (p<0.05) in 11 bacterial species, including a 5.6-fold increase in L. johnsonii, a bacteria that has been shown to improve metabolic outcomes in DIO models. In contrast, though OEA reduced body weight, OEA treatment had relatively little effect on the microbiome. To determine whether Ex4 and OEA directly alter the gut bacteria profile, we cultured gut bacteria isolated from DIO mice and treated the cultures with Vehicle, OEA (10µM), or Ex4 (10nM) for 24 hours. 16S analysis of in vitro cultures revealed no change in bacteria profiles for either treatment. These results suggest that Ex4 indirectly alters the gut microbiome to promote weight loss. Future experiments will determine if microbiome alterations contribute to the Ex4-mediated elevation in EE and improvements in metabolic outcomes.

Disclosure

J. Brown: None. V. Sharma: None. D. Tran: None. S.N. Peterson: None. J. Ayala: None. J. Brown: None.
Iron Downregulates Leptin by Decreasing CREB O-GlcNAc Modification and CREB Occupancy of the Leptin Promoter
Authors: LIU JINGFANG , GAO YAN , LORENZO FELIPE , SINK SANDY T. , MCCLAIN DON
Published: June 2018

Modification of proteins by O-linked N-acetylglucosamine (O-GlcNAc) serves as a nutrient-dependent signal to regulate transcription of the leptin gene. Recently we reported that increased levels of iron, which are also a risk factor for type 2 diabetes, down regulate leptin in mice fed high iron diets (2000 mg/kg) or in 3T3L1 cells treated with iron. Iron increased cyclic AMP-dependent response element binding protein (CREB) phosphorylation and its occupancy at inhibitory sites in the leptin promoter, but without demonstrable increases in CREB kinase activities. We now demonstrate that nutrient- and iron-dependent regulation of leptin are linked through O-GlcNAc modification of CREB. CREB O-GlcNAc modification was decreased in mice fed high iron, and this was associated with 54±7% and 44±9% reductions in recruitment to its two sites in the leptin promoter as analyzed by ChIP (p<0.05). Increasing levels of O-GlcNAc through targeted deletion of the O-GlcNAcase (OGA) gene in adipocytes increased adipose leptin mRNA expression by 86±12% (p<0.05) in mice fed a low-normal iron diet (35 mg/kg) and partially rescued the effect of high iron (35±3%, p<0.05). Similar effects were seen in serum leptin levels: High iron caused a 72% decrease and OGA deletion resulted in a rescue of 34%. Treatment of 3T3L1 cells with iron decreased expression of a leptin reporter plasmid by 45±4% and glucosamine, which bypasses the rate-limiting step in synthesis of UDP-GlcNAc, the substrate for O-GlcNAcylation, increased protein O-GlcNAc and fully rescued (106±5%) the effect of high iron (p<0.001 for both). Although iron also decreased global levels of protein O-GlcNAc modification, mRNA levels of the enzymes that control the modification, namely O-GlcNAc transferase and OGA, were unchanged. We conclude that adipose tissue integrates cellular nutrient status and tissue iron levels to control the regulation of leptin, through regulation of O-GlcNAc modification of the transcription factor CREB.

Disclosure

J. Liu: None. Y. Gao: None. F. Lorenzo: None. S.T. Sink: None. D. McClain: None.
Peripheral Melanocortin 3 Receptor (MC3R) Regulates Hepatic Autophagy in Obesity
Authors: JUN JOO YUN , SEO ARNOLD Y. , PATEL TUSHAR P. , UHLMAN ANDREW J. , LEVI NOAH J. , ROBERSON ROBIN , YANOVSKI JACK A.
Published: June 2018

We have previously reported homozygosity for two MC3R SNPs (C17A+G241A) that partially inactive MC3R increases adiposity in humans and mouse models. However, the precise mechanisms by which hypoactive MC3R alters metabolic homeostasis remain unclear. Given the essential role of hepatic autophagy in cellular mechanisms of obesity, we hypothesized that diminished peripheral MC3R activity leads to hepatic lipid accumulation by interfering with autophagy, leading to steatosis. We used knock-in mouse models that carry either homozygous human “wild type” (MC3RhWT/hWT) or “double mutant” MC3R (C17A+G241A, MC3RhDM/hDM). We studied hepatic autophagic pathways with biochemical/imaging analysis in wild type Mc3r (C57/BL6) and MC3RhWT/hWT as controls (WT mice), and in MC3RhDM/hDM, and mice lacking Mc3r (Mc3r-/-) which each have greater fat mass and hepatic triglycerides. We first demonstrated the MC3R specific agonist D-trp-γ-MSH induced LC3II (a marker of autophagosome formation) in hepatocytes from WT, but not Mc3r-/- or MC3RhDM/hDM mice. Interestingly D-trp-γ-MSH also induced TFEB (a master regulator of lysosomal biogenesis) translocation from cytosol to nucleus in TFEB-overexpressed NIH-3T3 cells, indicating MC3R may regulate autophagy by affecting TFEB signaling. Overnight fasting induced liver LC3II expression in WT mice, but not in MC3RhDM/hDM or Mc3r-/-; rather, basal LC3II was upregulated even in the fed state for the latter two genotypes. Hepatocytes from MC3RhDM/hDM or Mc3r-/- failed to increase autophagic flux with starvation or with the presence of chloroquine, suggesting MC3R may play a role in the regulation of autolysosomal degradation under the control of TFEB. Electron microscopy analysis of liver sections further demonstrated MC3RhDM/hDM mice exhibited fewer lipid droplet-associated autophagosomes compared to MC3RhWT/hWT mice. These data suggest MC3R plays an important peripheral role for the regulation of hepatic autophagy that is targeted to lipids.

Disclosure

J. Jun: None. A.Y. Seo: None. T.P. Patel: None. A.J. Uhlman: None. N.J. Levi: None. R. Roberson: None. J.A. Yanovski: Research Support; Self; Rhythm Pharmaceuticals Inc., Zafgen.
Dietary Animal and Saturated Fat Predict Future Visceral Fat Accumulation in Japanese Americans
Authors: HAYASHI TOMOSHIGE , SATO KYOKO , LEONETTI DONNA L. , KAHN STEVEN E. , UEHARA SHINICHIRO , FUJIMOTO WILFRED Y. , BOYKO EDWARD J.
Published: June 2018

Both diet and visceral adiposity play key roles in cardiometabolic disease. However, the association between dietary fat and subsequent visceral fat accumulation is not known. Thus, we prospectively examined future visceral fat accumulation in relation to dietary intake of animal and plant fats, or saturated and polyunsaturated fats in 312 nondiabetic Japanese-American men and women. Visceral adiposity was measured as intra-abdominal fat (IAF) area at the umbilicus by computed tomography at baseline and 10-11 years. Detailed dietary information was assessed at baseline using a food frequency questionnaire and 24-hour recall. Insulin sensitivity was evaluated by homeostasis model assessment for insulin resistance (HOMA-IR) and insulin response by the insulinogenic index (IGI) [Δinsulin/Δglucose (30-0 minutes)]. After adjustment for baseline IAF, age, sex, smoking habits (non-smokers, past-smokers, current smokers), physical activity, alcohol consumption, HOMA-IR, IGI, total protein intake, and total carbohydrate intake in multiple linear regression models, baseline log transformed animal fat intake [coefficient=15.319 (P=0.002)], but not plant fat intake [0.163 (P=0.270)], was associated with future IAF change. In the model that included saturated and polyunsaturated fat substituted for animal and plant fat intake, saturated fat was associated with future IAF change in both men [0.892 (P=0.030)] and women [1.121 (P=0.022)], but a statistically significant interaction was present between sex and polyunsaturated fat, such that polyunsaturated fat intake was inversely associated with future IAF change in men [-1.290 (P=0.003)], but not women [-0.058 (P=0.905)].

In conclusion, in both men and women, higher intake of animal fat and saturated fat, not plant fat, precede more visceral fat accumulation, while in men only, higher polyunsaturated fat intake precedes less visceral fat accumulation.

Disclosure

T. Hayashi: None. K. Sato: None. D.L. Leonetti: None. S.E. Kahn: Advisory Panel; Self; Boehringer Ingelheim GmbH, Elcelyx Therapeutics, Inc., Eli Lilly and Company, Intarcia Therapeutics, Inc., Janssen Research & Development, Merck & Co., Inc., Novo Nordisk A/S. S. Uehara: None. W.Y. Fujimoto: None. E.J. Boyko: None.
APPL2 Promotes Inflammation in Mouse Liver by Negative Regulation of Adiponectin Signaling
Authors: RYU JIYOON
Published: June 2018

Adiponectin is an insulin sensitizer that exerts antidiabetes and anti-inflammation functions. Adiponectin signaling is negatively regulated by the adaptor protein APPL2 in cells, yet the in vivo roles of APPL2 remains largely unknown. In this study, we show that liver-specific knockout of APPL2 improved insulin sensitivity and enhanced adiponectin signaling in high fat diet (HFD)-fed mice. The liver-specific APPL2 knockout mice (APPL2LKO) had less macrophage infiltration in liver tissues compared with their wild type controls and were resistant to LPS-induced expression of inflammatory genes such as IL-6, IL-1α, MCP-1, and TNF-α. Pretreatment of macrophage Raw264.7 cells with conditioned media from primary hepatocytes of APPL2LKO mice attenuated LPS-stimulated expression of inflammatory genes and the migration of the cells. The anti-inflammatory effect of APPL2 knockout was significantly reduced in adiponectin and liver-specific APPL2 double knockout mice (AdiponectinKO/APPL2LKO). Taken together, our results show that APPL2 promotes inflammation in mouse liver and the protein exerts a pro-inflammatory role by antagonizing adiponectin signaling.

Disclosure

J. Ryu: None.
The Transcription Factor ETV5 Regulates Adipogenesis through PPARg Signaling Pathway
Authors: MAO ZHUO
Published: June 2018

ETV5 is a member of PEA3 subfamily of E26 transformation-specific (ETS) transcription factors, characterized by the conserved DNA binding domain, ETS domain. Recently, a series of genomic-wide association studies (GWAS) found that ETV5 is associated with obesity. ETV5 knockout mice have lower fat mass and resistant to diet induced obesity, suggesting it was involved in the adipogenesis. In our study, we showed that ETV5 was expressed in the nucleus of adipocyte. Its expression level was altered according to different nutrient status. In order to exclude other tissues’ effect, we examine its biological function in preadipocyte 3T3-L1 cells. We found that the protein level but not the mRNA level of ETV5 was significantly elevated during adipogenesis. We utilized lentivirus to successfully knockdown ETV5 in 3T3-L1 cells. Deficiency of ETV5 resulted in reduced number and smaller size of adipocytes showed by oil red O staining. The major genetic changes involved in the impaired adipogenesis was defective PPAR signaling. Rosiglitazone stimulation of 3T3-L1 could restore adipogenesis caused by ETV5 deficiency. Dual luciferase assay found that ETV5 potently enhanced PPRE activity. These results suggest ETV5 is an important regulator of adipogenesis in early stage and a potential target for the treatment of obesity.

Disclosure

Z. Mao: None.
The Vagus Nerve Mediates Severe Hypoglycemia–Induced Fatal Cardiac Arrhythmias
Authors: RENO CANDACE M. , SKINNER ALLIE J. , BAYLES JUSTIN , FISHER SIMON J.
Published: June 2018

Severe hypoglycemia causes fatal cardiac arrhythmias. The extent to which the parasympathetic nervous system regulates hypoglycemia-induced fatal arrhythmias remains unknown. To investigate the role of vagal innervation in regulating arrhythmias during severe hypoglycemia, Sprague Dawley rats underwent vagotomy (left vagus nerve transection adjacent to the carotid artery; n = 15) or sham surgery (control; n = 13). One week after surgery, all rats underwent hyperinsulinemic (0.2 U/kg/min) severe hypoglycemic (10-15 mg/dl) clamps with electrocardiogram recordings. Rats with vagotomy had increased heart rate at baseline and throughout the clamp, indicating successful vagotomy. Glucose infusion rates during hypoglycemia were increased 1.7-fold in rats with vagotomy, but epinephrine values were similar between the groups. Vagotomy resulted in fewer cardiac arrhythmias including a 12-fold decrease in 2nd degree heart block. Reduction in arrhythmias was associated with a 7-fold decrease in severe hypoglycemia-induced mortality in rats with vagotomy (7%) compared to controls (46%).

In summary, vagal transection reduces mortality and cardiac arrhythmias during severe hypoglycemia. Since vagal innervation mediates severe hypoglycemia-induced sudden death, targeting the parasympathetic nervous system may be a logical approach to prevent sudden death for at-risk people with insulin treated diabetes.

Disclosure

C.M. Reno: None. A.J. Skinner: None. J. Bayles: None. S.J. Fisher: None.
Sennoside A Improves Mitochondrial Function of Large Intestine in Diet-Induced Obese Mice to Preserve GLP-1 Expression through Induction of Short-Chain Fatty Acids
Authors: LE JIAMEI , YE JIANPING , SUN YONGNING
Published: June 2018

Objective: Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative. However, its activity remains unknown in the regulation of GLP-1 expression. Here, we investigated the possibility in high-fat-diet (HFD) induce obesity (DIO) mice.

Methods: SA was administrated in DIO mice through dietary supplementation for 8 weeks. In the study, energy balance and insulin sensitivity were monitored. GLP-1 was determined in plasma and colon tissue. Mitochondria were examined in structure by transmission electron microscopy and in function with Seahorse XF24 in the colon tissue. Gut microflora was examined using fecal DNA and the diversity of gut microbiota was analyzed by Miseq sequencing. The fecal short chain fatty acids (SCFAs) were examined by GC (gas chromatograph).

Results: There was an increase in cell necrosis in the epithelial layer of mucosa of DIO mice, which was observed with reduction of plasma GLP-1 and mRNA expression in the colon tissue. Mitochondria exhibited an increase in size and membrane rupture in the epithelial cells. An elevation in the complex I activity, and a fall in the complex II activity were observed in the mitochondria, suggesting an imbalance of the complex I/II in the cells. The short chain fatty acids were decreased in abundance along with the dysbiosis in the colon of DIO mice. All of those alterations were corrected by SA in DIO mice along with control of obesity and insulin resistance.

Conclusion: The data suggested that SA may improve GLP-1 expression in colon through restoration of mitochondrial function and structure in the epithelia cells of mucosal. The effect may be related to restoration of gut microflora/SCFAs axis.

Disclosure

J. Le: None. J. Ye: None. Y. Sun: None.
Early Life Exposure to Antibiotic Alters Energy Balance during Chronic High-Fat Feeding in Adult Male and Female Mice
Authors: NOH HYE LIM , SUK SUJIN , FRIEDLINE RANDALL H. , INASHIMA KUNIKAZU , TRAN DUY A. , KIM ALLISON M. , KIM SOOYOUNG , JEONG SANGEUN , UONG CECILIA , OZKAN BERK A. , KASINA VISHAL , KNOWLES KATHERINE P. , PEREZ-PEREZ GUILLERMO , LEE KI WON , BLASER MARTIN J. , KIM JASON K.
Published: June 2018

Gut microbiota are affected by antibiotics and may modulate host energy metabolism. Male and female C57BL/6J mice (n=5∼11/group) were exposed to low-dose penicillin (LDP) via maternal drinking water from birth to 4 weeks (w) of age, and chronic changes in energy balance were examined using metabolic cages. Through 8w of age (Chow), LDP and Control mice had comparable fat mass, but male and female LDP mice showed significantly increased O2 consumption rates without changes in food intake and activity (Figure). At 9w of age, mice were given a high-fat diet (HFD; 45% fat by calories), and they began gaining adiposity with female LDP mice showing ∼40% lower fat mass than Controls. After 8w of HFD, male LDP group showed significantly elevated O2 consumption rates compared to Controls. Male LDP mice also showed marked reductions in food intake and activity, but these effects were no longer observed after 12w of HFD. In contrast, female LDP group had reduced O2 consumption rates compared to Controls after 4 and 8w of HFD without changes in food intake and activity.

In conclusion, these results indicate that early life exposure to penicillin modulates energy balance in young and mature mice, suggesting an important role of altered populations of gut microbiota in diet-induced obesity. Our findings also identify important gender-selective effects of early life antibiotic exposure on energy balance in mature mice.

Disclosure

H. Noh: None. S. Suk: None. R.H. Friedline: None. K. Inashima: None. D.A. Tran: None. A.M. Kim: None. S. Kim: None. S. Jeong: None. C. Uong: None. B.A. Ozkan: None. V. Kasina: None. K.P. Knowles: None. G. Perez-Perez: None. K. Lee: None. M.J. Blaser: None. J.K. Kim: None.
Genome-Wide and Abdominal Imaging Data Characterizes Common Alleles Associated with Higher BMI and Subcutaneous Fat but Less Liver Fat and Lower Risk of Type 2 Diabetes
Authors: YAGHOOTKAR HANIEH , JI YINGJIE , YIORKAS ANDRIANOS M. , FRAU FRANCESCA , MOOK-KANAMORI DENNIS , MUTSERT RENÉE DE , TYRRELL JESSICA , JONES SAMUEL E. , BEAUMONT ROBERT , WOOD ANDREW R. , THOMAS LOUISE , ALLEBRANDT KARLA V. , STEFAN NORBERT , STAIGER HARALD , PEARSON EWAN , BLAKEMORE ALEX I. , BELL JIMMY D. , FRAYLING TIMOTHY M.
Published: June 2018

Genetic studies have identified “favourable adiposity” variants - where the allele associated with higher adiposity is associated with lower risk of type 2 diabetes. We took a novel approach to find more of these alleles and find the underlying mechanisms.

We first performed a genome wide association study (GWAS) of body fat percentage using 451000 individuals from UK Biobank. Second, we used published genetic data in a multivariate test to find alleles associated with higher adiposity but a “favourable” metabolic phenotype: higher HDL-C, adiponectin, sex hormone binding globulin, but lower triglycerides, fasting insulin and alanine transaminase. Third, we used abdominal imaging data from 4 studies to define the adiposity phenotype in more detail.

We identified 620 variants associated with body fat percentage (p<5x10-8). Fourteen alleles had a “favourable adiposity” phenotype, including 7 previously known variants (in/near PPARG, LYPLAL1, GRB14, IRS1, PEPD, FAM13A and ANKRD55), 5 known to be associated with a relevant trait (TRIB1, KLF14/MKLN1, DNAH10, VEGFA/C6orf223 and AEBP2/PDE3A) and 2 novel variants (MAFF and CITED2). Carrying 10 additional "favourable adiposity" allele was associated with higher BMI (0.63 kg/m2, p = 4x10-45) but lower risk of type 2 diabetes (odds ratio: 0.60, p = 4x10-44, 14371 cases), lower risk of heart disease (0.82, p = 3x10-14, 37741 cases) and lower risk of hypertension (0.82, p = 1x10-33, 241691 cases). Using MRI/CT scans of abdominal fat from UK Biobank (N = 5000), NEO (2236), TÜF (900), and a published GWAS (10557), carrying more “favourable adiposity” alleles was associated with higher subcutaneous fat (p = 1x10-12) but lower liver fat (p = 1x10-7). These effects were similar when we removed the 7 known “favourable adiposity” variants.

“Favourable adiposity” alleles associated with higher BMI but lower risk of type 2 diabetes are characterized by higher subcutaneous but lower liver fat.

Disclosure

H. Yaghootkar: None. Y. Ji: None. A.M. Yiorkas: None. F. Frau: None. D. Mook-Kanamori: None. R. de Mutsert: None. J. Tyrrell: None. S.E. Jones: None. R. Beaumont: None. A.R. Wood: None. L. Thomas: None. K.V. Allebrandt: None. N. Stefan: Consultant; Self; Merck Sharp & Dohme Corp.. Speaker's Bureau; Self; Novo Nordisk A/S. Research Support; Self; AstraZeneca. Speaker's Bureau; Self; AstraZeneca, OmniaMed Ltd.. H. Staiger: None. E. Pearson: Speaker's Bureau; Self; Eli Lilly and Company. A.I. Blakemore: None. J.D. Bell: None. T.M. Frayling: None.