bluuebunny/crossref_metadata_2025 huggingface dataset:
1737-P: Prevalence of Autoimmune Diseases in Longstanding Diabetes: Results from the Canadian Study of Longevity in Type 1 Diabetes
Authors: CARDINEZ NANCY , LOVBLOM LEIF ERIK , ORSZAG ANDREJ , CHERNEY DAVID , PERKINS BRUCE A.
Published: June 2019

Objective: Type 1 diabetes (T1D) predisposes to higher lifetime autoimmune (AI) disease risk; the magnitude of this risk is not known with certainty. To approximate lifetime risk, we aimed to determine the prevalence of AI diseases in those exceeding 50 years T1D duration, as well as to explore sex-specific differences and associations with complications.

Methods: Descriptive analysis of the Canadian Study of Longevity in T1D, a nationwide questionnaire study initiated in 2013 that recruited 374 T1D participants with duration >50 years. The questionnaire collected diabetes management variables, general medical history with a focus on AI disease, and diabetes-related complications. Clinical and laboratory data were obtained from primary care providers. The χ2-test and Student’s t-test were used for simple comparisons.

Results: The 374 participants with longstanding T1D had median age of 66 [59, 71] years, T1D duration 53 [51, 58] years and HbA1c 7.5±1.0%. AI diseases were reported in 184/374 (49.3%). Thyroid disease was most prevalent, 154/374 (41.2%) reported thyroid disease or was prescribed thyroid hormone replacement therapy. Addison’s disease or prescribed glucocorticoid and mineralocorticoid therapy was reported in 5/374 (1.3%). Autoimmune polyendocrine syndrome type 2 (APS-2), defined by the presence of thyroid disease, Addison’s disease and T1D, was present in 3/374 (0.8%). We observed a sex-specific difference in AI disease prevalence (34.6 females vs. 14.8% in males; p<0.0001), driven by the difference in thyroid disease (29.5 females vs. 11.5% males, p<0.0001). No difference in HbA1c was observed (7.5±1.1 vs. 7.6±1.0%, p=0.62). There was a trend toward lower CAD prevalence among those with AI disease (12.1 vs. 16.9%, p=0.05).

Conclusions: We estimate that in a population with longstanding T1D, the lifetime risk of AI disease approaches half, driven primarily by the presence of autoimmune thyroid disease, and with greater risk in females.

Disclosure

N. Cardinez: None. L. Lovblom: None. A. Orszag: None. D. Cherney: Other Relationship; Self; AbbVie Inc., AstraZeneca, Bayer AG, Boehringer Ingelheim International GmbH, Janssen Pharmaceuticals, Inc., Merck & Co., Inc., Mitsubishi Tanabe Pharma Corporation, Prometic Life Sciences Inc., Sanofi. B.A. Perkins: Advisory Panel; Self; Abbott, Boehringer Ingelheim International GmbH, Boehringer Ingelheim International GmbH, Insulet Corporation. Research Support; Self; Boehringer Ingelheim International GmbH. Other Relationship; Self; Abbott, Boehringer Ingelheim International GmbH, Lilly Diabetes, Medtronic, Novo Nordisk Inc., Sanofi.

Funding

JDRF (17-2013-312)
1752-P: GDF11 Involved in Expressed MicroRNAs during GLP-1RA Inhibited Adipogenesis
Authors: XUE PENG , WANG NA , LI ZIYI , LI SHILUN , YUKUN LI
Published: June 2019

Differentiation of bone marrow mesenchymal stem cells (BMSCs)is controlled by a vast array of mechanisms, with miRNAs-mRNAs regulation network identified widely recently. In this study, we aimed to specify the differentially expressed (DE) miRNAs after glucagon-like peptide-1 receptor agonist (GLP-1RA) administration during adipogenesis in BMSCs. MiRNAs extracted from three different groups, [the control group (C), the adipogenesis group (A), and the GLP-1RA administration group (GA)], were sent for high-throughput sequencing. We identified 5 DE miRNAs (miRNA-150-5p, miRNA-129-5p, miRNA-201-3p, miRNA-201-5p, and miRNA-214-5p) among groups. The expression of the 5 DE miRNAs were verified by the real-time RT-PCR analysis and consistent with the small-RNA sequencing data. Target genes were then predicted, and highly enriched GOs and KEGG pathway analyses indicated that they were involved in multiple biological processes concerning adipogenesis. Two of the target genes, growth differentiation factor 11 (GDF11) and cholecystokinin B receptor (CCKBR), were verified for functional study. Actually, the mRNA levels of GDF11, rather than CCKBR, was down-regulated by miRNA-150-5pand involved in GLP-1RA administration. Taken together, these results suggested for the first time that GDF11 was involved in differentially expressed microRNAs during GLP-1RA inhibited adipogenesis in primary BMSCs.

Disclosure

P. Xue: None. N. Wang: None. Z. Li: None. S. Li: None. L. Yukun: None.

Funding

Natural Science Foundation of Hebei Province (H2016206243); Department of Health of Hebei Province (361005)
1755-P: Transferrin Role in Systemic Iron Influence on Insulin Sensitivity and Adipocyte Function
Authors: LORENZO FELIPE , JAIN SHALINI , MCCLAIN DON , DAS SWAPAN K.
Published: June 2019

Systemic iron overload is associated with decreased insulin sensitivity (Si) and diabetes. We previously characterized the genetic regulation of iron and its effect on Si in humans, which demonstrated that expression of the iron transport protein transferrin (Tf) in adipose tissue plays a role in regulating insulin sensitivity. To further characterize this effect at the tissue and cellular levels, we generated heterozygous Tf knock out (KO) mice and human adipose cells (ASC52Telo) with knock down (KD) of Tf. Heterozygous Tf KO and wild type (WT) mice were fed a 40% fat “fast-food” diets, supplemented with glucose in the water, containing either 35mg or 2000mg iron per kg chow. Fasting glucose levels were not affected in the Tf KO mice compared to WT (p=0.3889) on the low iron diet, but increased in the KO mice on 2gm/kg iron (13% increase, p=0.0511) suggesting the attenuation of the protective effect of transferrin in blood sugar regulation at high iron diet. We previously reported that adipose cells transfected with TF shRNA compared to a control vector had decreased intracellular iron and exhibited insulin resistance. We also performed mitochondrial function studies (oxygen consumption analyzed by Seahorse) in these cells. Tf KD cells exhibited decreased maximal oxygen consumption compared to controls (p:0.0004). Iron treatment of the cells augmented respiration in the control cells but worsened maximal oxygen consumption in the Tf KD cells (p=0.0011). We conclude that transferrin-mediated iron transport in adipocytes plays an important role in determining insulin sensitivity, allowing beneficial responses to high iron, namely protection from hyperglycemia and augmented mitochondrial function.

Disclosure

F. Lorenzo: None. S. Jain: None. D. McClain: None. S.K. Das: None.

Funding

U.S. Department of Veterans Affairs
1772-P: Insulin Granule Biogenesis at the Golgi Requires a Metabolically-Regulated, Non-centrosomal Subpopulation of Microtubules
Authors: TROGDEN KATHRYN , WRIGHT CHRISTOPHER V. , GU GUOQIANG , KAVERINA IRINA
Published: June 2019

Insulin undergoes biogenesis through the ER and Golgi, ultimately leading to packaging into vesicles that are secreted in response to glucose. The movement between these membranes and the process of budding from the Golgi are vital steps. Previous work has found a role for microtubules (MTs) in these processes in both pancreatic β-cells and other cell types. Here, we show that the last stages of this process are regulated by a specific subpopulation of MTs that are non-centrosomal. Their formation, or nucleation, at the Golgi membrane is regulated by a glucose signal-transduction pathway through cAMP and its effector EPAC2. This effect is especially pronounced during the first, rapid phase of insulin secretion, during which EPAC2 levels increase at the sites of their nucleation. Preventing new nucleation of this specific population dramatically affects the pipeline of insulin production, storage and release. There is an overall reduction of β-cell insulin content, and the remaining insulin becomes retained within the Golgi, likely because of stalling of insulin-granule budding. This diminished granule availability substantially effects the level of both basal and stimulated insulin secretion, though not the ratio between these levels during the first wave. Constant dynamic maintenance of this network is therefore critical for normal β-cell physiology. This is the first demonstration that the biogenesis of post-Golgi carriers, particularly large secretory granules, requires ongoing nucleation/replenishment of this network. We are further looking at the role of MTs in β-cell secretory activity in intact islets. We have observed that MTs promote β-cell heterogeneity by restricting secretion from a specific β-cell sub-population, where insulin release can be activated upon disruption of the entire MT network by the drug nocodazole.

Disclosure

K. Trogden: None. C.V. Wright: None. G. Gu: None. I. Kaverina: None.

Funding

National Institutes of Health (1F32DK117529-01A1)
1777-P: Acute Hyperbaric Oxygen (HBO) Treatment Increases Hepatic Insulin Sensitivity and Muscle Reactive Oxygen Species in Type 2 Diabetes
Authors: SARABHAI THERESIA , ANADOL-SCHMITZ EVRIM , KATSUYAMA HISAYUKI , MARKGRAF DANIEL F. , BUSCH LUCAS , GUTHOFF RAINER , SCHNEPPENDAHL JOHANNES , RODEN MICHAEL
Published: June 2019

Diabetes mellitus type 2 (T2D) is characterized by insulin resistance of liver and skeletal muscle, which is at least partly due to impaired muscle mitochondrial function. Long-term HBO therapy, as applied for treating the diabetic foot syndrome, has been shown to improve blood glucose concentrations. To study the underlying mechanisms, we examined the short-term effect of HBO treatment on insulin sensitivity and mitochondrial oxidative capacity and production of reactive oxygen species (ROS) in a randomized, double blinded, placebo-controlled trial. Volunteers with type 2 diabetes (n=4; age 61±4 years; body mass index 30.2±2.2 kg/m2; HbA1c 7.0±0.7%) underwent two 2-hours sessions in a hyperbaric chamber of once with application of 100% oxygen (1.4 bar over atmospheric pressure [barO] for 20 min; HBO) and once with ambient air as placebo (21% oxygen; 1.4 barO; PLC). Tissue-specific insulin sensitivity was assessed by hyperinsulinemic-euglycemic clamps with stable isotope dilution, while muscle oxygen fluxes were measured with high resolution respirometry. After HBO treatment, hepatic insulin sensitivity was higher (insulin-mediated suppression of endogenous glucose production: 69.7±0.1% vs. PLC 55.3±3.2%, p<0.05), whereas peripheral insulin sensitivity was comparable (2.0±0.3 vs. 2.2±0.3 mg*kg-1*min-1, p=0.24). Muscle ROS production markedly increased after HBO (increase in ROS: 73±3% vs. PLC -14±5%; p<0.005), while mitochondrial oxidative capacity only tended to be lower after HBO.

In conclusion, a single session of HBO treatment already improves hepatic insulin resistance and stimulates muscle ROS release, which may activate antioxidative defense, both of which could contribute to improved glycaemia upon chronic treatment.

Disclosure

T. Sarabhai: None. E. Anadol-Schmitz: None. H. Katsuyama: None. D.F. Markgraf: Research Support; Self; Sanofi. L. Busch: None. R. Guthoff: None. J. Schneppendahl: 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.
179-OR: Low-Dose Dihydrotestosterone Lowers Lipogenic Master Regulator in Liver and Adipose Tissue from Female Mice
Authors: ANDRISSE STANLEY
Published: June 2019

Hyperandrogenemia (HA) and insulin resistance (IR) are hallmarks of polycystic ovary syndrome (PCOS), a common endocrine disorder that affects 1 in 10 women. These hallmarks are also integral elements of nonalcoholic liver disease (NALFD), a disorder that is common in women with PCOS. Administering low dose dihydrotestosterone (DHT) induced a lean female mouse model with a PCOS-like phenotype, displaying IR and NAFLD. The molecular mechanism of HA-induced NAFLD has not been determined. We hypothesized that low dose DHT would interrupt hepatic lipid metabolism leading to NAFLD. To investigate the role of androgens on the master regulator of lipogenesis, sterol regulatory element-binding protein 1 (SREBP-1), we extracted white adipose tissue (WAT), liver, and skeletal muscle from wild type, control and low dose DHT female mice; and performed Western blot and real-time quantitative PCR (qRT-PCR) analysis of lipogenic intermediates of the tissue homogenates. Low-dose DHT lowered the active form of cytosolic SREBP1 in the liver and WAT compared to controls. Additionally, low-dose DHT lowered inactive SREBP1 in the liver. However, the condition did not alter the levels of the active and inactive forms of SREBP2 in the liver and WAT, though the active form was lowered in skeletal muscle. Further, p-ACC levels were unaltered in liver and WAT. FAS levels were unchanged in WAT and skeletal muscle. Taken together, our findings support the hypothesis that cytosolic SREBP1 decreased due to its translocation to the nucleus, where it regulates lipogenic protein levels. We speculate that low-dose DHT promotes the translocation of SREBP-1 from the cytosol to the nucleus to influence lipogenic gene expression leading to increased lipogenesis contributing to NAFLD.

Disclosure

S. Andrisse: None.
1795-P: Novel Techniques for the Analysis of Brain Chondroitin Sulfates in Rodents and Humans with Type 2 Diabetes
Authors: ALONGE KIMBERLY M. , LOGSDON ARIC , BANKS WILLIAM A. , KEENE C. DIRK , SCHWARTZ MICHAEL W. , GUTTMAN MIKLOS
Published: June 2019

We recently reported that perineuronal nets (PNNs), which are chondroitin sulfate (CS) containing matrices that function to maintain synaptic integrity of key neurocircuits, are present in the hypothalamus of mice, rats and humans (Mizadeh et al, Nat Metabolism, 2019). AgRP neurons are among those enmeshed by PNNs in this brain area and excessive activation of these neurons is suggested to play a role in the pathogenesis of T2D. Our finding that PNNs are abnormal in the hypothalamus of T2D rodents offers a plausible link between brain mediated glucose-sensing dysfunction and T2D pathogenesis. PNN matrix stability correlates closely with the abundance of specific CS variants that constitute the PNN matrix, with an increase in 6S-CS promoting matrix destabilization. Traditional methods for CS-based ECM analysis in fixed brain tissues depend primarily upon IHC detection of CS-specific disaccharides, but this method is limited by cross-reactivity of CS-specific antibodies. Here we report the development and validation of two novel and quantitative biochemical methods for the in-depth compositional analysis of four biologically relevant CS-derived disaccharides present in rodent and human fixed brain tissue: 1) electrospray ionization-tandem mass spectrometry (MS/MS) to determine the percentages of 4S- and 6S-CS within the monosulfated population, and 2) liquid chromatography (LC)-MS/MS for resolving and determining percentages of 0S- 4S- 6S- and 4S6S-CS. These methods can detect CS from fixed coronal brain slices with nanomolar sensitivity and high reproducibility. After optimization using synthetic standards, both methods were validated using a developmental time course series in fixed rodent brain tissue and then applied to the analysis of formalin-fixed human brain tissue. These methods offer a useful approach with which to investigate the role of aberrant CS composition in the pathogenesis of diabetes and related metabolic disorders.

Disclosure

K.M. Alonge: Research Support; Self; Novo Nordisk Inc. A. Logsdon: Research Support; Spouse/Partner; Novo Nordisk Inc. W.A. Banks: None. C. Keene: None. M.W. Schwartz: Consultant; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S. M. Guttman: None.

Funding

National Institutes of Health; Novo Nordisk
1808-P: Is There Evidence of Gliosis Outside the Mediobasal Hypothalamus in Obese Adults?
Authors: ROSENBAUM JENNIFER L. , MELHORN SUSAN J. , DE LEON MARY ROSALYNN , WEBB MARY , SCHUR ELLEN
Published: June 2019

Data increasingly support evidence of gliosis in the mediobasal hypothalamus (MBH) in the setting of obesity and insulin resistance. It is if other regions of the brain are similarly affected. We tested if MBH gliosis is related to radiologic evidence of gliosis in selected subcortical, cortical, and hypothalamic regions.

We recruited 40 obese adults (aged 25-61 y), with variable glucose tolerance by OGTT (n=24 normal glucose tolerance, n=11 impaired glucose tolerance and n=5 with T2DM). Individuals underwent a multi-echo T2 brain MRI. Two analysts placed regions of interest (ROIs) on the scans and determined the T2 relaxation time as a measure of gliosis. ROIs included the MBH, amygdala, dorsomedial hypothalamus (DMH; area of the paraventricular nucleus), hippocampus, white matter (WM), cortex, caudate, ventral striatum (VS), and putamen. Left and right sides were averaged. High (n=13) and low (n=14) T2 groups were derived from the tertiles with the highest (strongest evidence for gliosis) and lowest T2 relaxation times. The groups were compared by linear regression adjusted for age and sex.

Mean BMI in the low T2 group was 35.7 ± 4.1 kg/m2 and high T2 group was 37.1 ± 4.9 kg/m2. The groups did not differ in T2 relaxation time of WM (P=0.29), cortex (P=0.15), caudate (P=0.49), putamen (P=0.79), amygdala (P=0.09), or hippocampus (P=0.44). In the high T2 group, T2 relaxation time was shorter in the VS (adjusted mean 80.9±2.9 vs. 83.1±3.0, P=0.04) and longer in the DMH (adjusted mean 108.9±5.3 vs. 103.7±5.4, P=0.02), compared to the low T2 group. When subjects with T2DM (n=1 in low and n=3 in high T2 group) were excluded, the differences were attenuated (VSP=0.13; DMHP=0.10).

The findings reveal preliminary evidence of longer T2 relaxation times in the DMH of obese individuals with signs of MBH gliosis and no evidence for gliosis in cortical or subcortical regions. Further study of gliosis in the DMH and its relation to blood glucose control and autonomic function in humans, particularly in those with T2DM, is warranted.

Disclosure

J.L. Rosenbaum: None. S.J. Melhorn: None. M. De Leon: None. M. Webb: None. E. Schur: None.

Funding

American Diabetes Association (1-17-ICTS-085 to E.S.); National Institutes of Health (T32HL007028); University of Washington Institute of Translational Health Sciences (UL1TR002319)
1820-P: Autonomic Tone Is Necessary for Bile Acid-Induced Hepatic Insulin Resistance
Authors: SYRING KRISTEN , MIGNEMI NICHOLAS A. , MCGUINNESS OWEN P.
Published: June 2019

Diabetes and obesity cost the American healthcare system in excess of $322 billion dollars annually. Currently, one of the most effective treatments for obesity and metabolic disorders is bariatric surgery. Bariatric surgery leads to almost immediate metabolic benefits, independent of weight loss. Concurrent with the metabolic benefits, an elevation in systemic bile acids is also detected in these patients. Several investigations posit that the rise in systemic bile acids contributes to, or potentially drives, the beneficial outcomes described in these patients; however, the mechanism and direct effects of increased bile acids must be more thoroughly elucidated. Previous studies indicate that not only does the autonomic nervous system impact insulin action, but it also plays a role in bile acid synthesis and secretion. Specifically, studies in humans show that autonomic blockade in insulin resistant subjects leads to improved insulin sensitivity. Furthermore, in rats, a vagotomy results in increased serum bile acids concentrations. Prior data from our group demonstrates that during hyperinsulinemic-euglycemic clamps, increased circulating bile acids induce hepatic insulin resistance, resulting in a decrease in glucose infusion rates (GIR) (10±3 mg/kg/min) compared to control mice (21±4 mg/kg/min). Together these data led to the hypothesis that bile acids signal indirectly via the nervous system to impair hepatic insulin action. To test this hypothesis, autonomic tone was inhibited by infusion of a ganglionic blocker combined with an infusion of an alpha agonist to maintain blood pressure during clamps. In the presence of ganglionic blockade, bile acids do not induce hepatic insulin resistance. Instead, bile acid-treated mice have an increased GIR (66±3 mg/kg/min) compared to control mice (45±5 mg/kg/min). From these data we conclude that the autonomic nervous system is necessary for elevated systemic bile acids to impair insulin action in lean animals.

Disclosure

K. Syring: None. N.A. Mignemi: None. O.P. McGuinness: None.

Funding

National Institutes of Health
1849-P: Importance of Insulin’s Direct and Indirect Effects on Postprandial Hepatic Glucose Uptake
Authors: EDGERTON DALE S. , KRAFT GUILLAUME , FARMER BEN , SMITH MARTA S. , WILLIAMS PHILLIP E. , CHERRINGTON ALAN D.
Published: June 2019

In heathy individuals the liver is a primary site of glucose disposal, it is in a state of glucose uptake most of the day, and this process is impaired in diabetes. Insulin stimulates hepatic glucose uptake (HGU) but the importance of its indirect effects (at fat, brain and pancreatic alpha cells) in driving HGU is unclear. Thus, the aim of this study was to determine the impact of insulin’s indirect mechanisms of control of HGU. Postprandial conditions were simulated for 4-hour in conscious dogs (n=7/group) by infusing somatostatin, intraportal insulin (1.8 mU/kg/min) and glucose (4 mg/kg/min), and leg vein glucose (to clamp arterial glucose at 2-fold basal). In one group insulin’s direct and indirect effects on the liver were both present (D+I) while in the other insulin’s indirect effects were blocked (D-only). In D+I the basal intraportal glucagon infusion rate was decreased to create a progressive decline in glucagon over time, FFA levels were allowed to fall, and brain insulin action was allowed to increase. In D-only glucagon and FFA levels were maintained at basal levels by infusions of intraportal glucagon and leg vein lipid emulsion. Hypothalamic insulin action was blocked by 3rd ventricle infusion of an insulin receptor antagonist. In the D+I and D-only groups, respectively, hepatic insulin increased during the experimental period (from 20±3 to 88±8 and 21±3 to 89±8 µU/ml), hepatic glucagon fell or remained basal (39±5 to 24±3 and 40±6 to 41±2 pg/ml) and arterial FFA fell or remained basal (1069±19 to 99±23 and 1195±63 to 998±99 µmol/l). HGU and net hepatic glucose uptake (NHGU) were independently measured using tracer and cold glucose balance techniques. The AUCs for HGU and NHGU (mg/kg/240 min) in D+I were 841±143 and 792±105, respectively, and 738±130 and 698±58 in D-only (P=0.6 and 0.4 between groups, respectively). Thus, in the healthy animal, HGU responds normally to insulin even in the absence of the hormone’s indirect effects. Instead HGU is driven almost exclusively by insulin’s direct effects.

Disclosure

D.S. Edgerton: Speaker's Bureau; Self; Novo Nordisk A/S. G. Kraft: Consultant; Self; Metavention. B. Farmer: None. M.S. Smith: None. P.E. Williams: None. A.D. Cherrington: Board Member; Self; Biocon, Fractyl Laboratories, Inc., Metavention, Sensulin, LLC., vTv Therapeutics, Zafgen, Inc. Consultant; Self; Abvance, Boston Scientific Corporation, California Institute for Biomedical Research, Galvani Bioelectronics Limited, MedImmune, Novo Nordisk Inc., Thetis Pharmaceuticals LLC. Research Support; Self; Abvance, Diasome Pharmaceuticals, Inc., Novo Nordisk Inc. Stock/Shareholder; Self; Abvance, Biocon, Fractyl Laboratories, Inc., Metavention, Novo Nordisk Inc., Sensulin, LLC., Thetis Pharmaceuticals LLC, Zafgen, Inc.

Funding

National Institutes of Health
1850-P: Bromocriptine (B) plus Curcumin (C) Synergize to Reduce Nonalcoholic Steatohepatitis (NASH) in Mice
Authors: TSAI TSUNG-HUANG , CINCOTTA ANTHONY
Published: June 2019

NASH involves a combination of hepatic and gastrointestinal (GI) pathophysiologies comprising hepatic lipotoxicity and nitro/oxidative stress and a “leaky," proinflammatory GI environment that collude to induce the disease. B, a dopamine D2 receptor agonist is known to reduce plasma hyperFFAemia, and hepatic lipogenesis and inflammation. C has been demonstrated to reduce GI inflammation and leakiness, and is known to augment brain dopaminergic activity. We therefore investigated if B and C combination therapy that engages different pathological targets of NASH may provide an additive/synergistic benefit for NASH in a mouse model. C57B6 mice were maintained on 40 kcal% fat diet with 5% fructose drinking water for 20 weeks to induce NASH with fibrosis then randomized to 4 treatment groups: Vehicle, B (10mg/kg), C (100mg/kg), and combination of B and C (BC) (N=8/gr) daily for 4 weeks while maintained on the diet. A group of mice on normal diet was used as normal reference control. BC treatment led to significant reductions of hyperglycemia (21.8%), microbiome dysbiosis (38.1%), serum ALT (26.5%), obesity (10.1%), and liver steatosis (66.7%), inflammation (93.3%), ballooning (85.0%), NAFLD activity score (80.0%), as well as of four gene expression markers each of fibrosis (αSMA, TGFβ, COL1A, TXNIP) by 26-40%, inflammation (TNFa, IL1β, MCP1, iNOS) by 41-81%, ER stress (DDIT3, ATF, sXBP1, HSPA5) by 28-40%, steatosis (SREBP1c, FASN, ACC1, PLIN2) by 28-42%, and fatty acid oxidation (PPARα, PGC1α, CPT1, UCP1) by 24-84% and increased antioxidant gene expression (NQO, GPX1, CAT, HMOX1) by 29-70%, and reduced the percent fibrosis area by 80% relative to vehicle (and resultantly similar to normal diet mice) (all above: P<0.05) and by amounts greater than either B or C alone or additively. This is the first study to demonstrate the positive interactive effects of bromocriptine and curcumin on NASH in a mouse obesogenic dietary model of the disease and it offers a potential safe means of treating NASH.

Disclosure

T. Tsai: Employee; Self; VeroScience LLC. A. Cincotta: Employee; Self; VeroScience LLC.
1853-P: Transforming Growth Factor Beta 1 Acts as a Hepatokine in Control of Glucose and Energy Metabolism
Authors: PAN QUAN , CHEN YUNMEI , KIM DA MI , SHEN ZHENG , YANG WANBAO , LI XIAOPENG , SUN YUXIANG , GUO SHAODONG
Published: June 2019

Liver and adipose tissue are crucial for blood glucose and energy homeostasis, while impaired regulation of hepatic glucose production (HGP) and energy expenditure contribute to hyperglycemia and excessive fat storage in type 2 diabetes mellitus. Hepatokines are proteins secreted by hepatocytes, and several hepatokines have been shown to directly affect glucose and energy expenditure. In this study, we investigated the role of transforming growth factor beta 1 (TGF-β1) in hepatocytes in regulating glucose and energy homeostasis. We generated the liver-specific TGF-β1 knockout mice and found that the mice exhibited lower blood glucose, increased hepatic insulin sensitivity compared with wild type (WT) control mice. Using the mouse primary hepatocytes, we found that TGF-β1 deletion impaired HGP and TGF-β1 treatment promoted protein kinase A (PKA) signaling and stimulated the nuclear translocation of FoxO1. Under high fat diet (HFD) treatment, hepatic TGF-β1 deficiency protected mice from HFD-induced obesity and insulin resistance, decreased HGP and improved energy expenditure. Moreover, we used hepatic insulin receptor substrate-1, 2 (IRS1, 2) knockout (DKO) mice and deleted the TGF-β1 gene in the liver of DKO mice. We found that hepatic TGF-β1 ablation prevented hyperglycemia and hyperinsulinemia in DKO mice. Deletion of TGF-β1 or neutralization of TGF-β1 with antibody significantly decreased HGP of DKO hepatocytes. Additionally, hepatic TGF-β1 ablation enhanced thermogenesis and inguinal white adipose tissue (iWAT) browning in DKO mice. These findings underscore an important role of hepatic TGF-β1 in contribution to systemic glucose and energy homeostasis. Thus, hepatic TGF-β1 may serve as a therapeutic target for control of glucose and energy homeostasis in insulin resistance and type 2 diabetes mellitus.

Disclosure

Q. Pan: None. Y. Chen: None. D. Kim: None. Z. Shen: None. W. Yang: None. X. Li: None. Y. Sun: None. S. Guo: None.

Funding

American Diabetes Association (1-15-CD-09 to S.G.)
186-LB: Predictors of Diabetes Complication Screening at Two Years from Diagnosis in an Incident Cohort
Authors: AULICH JULIANE , CHAN ALBERT K. , CUSUMANO JANINE , PRYKE ALISON , BENITEZ-AGUIRRE PAUL Z. , CRAIG MARIA E. , DONAGHUE KIM C.
Published: June 2019

Background: Despite well-established national complication screening programmes for patients with diabetes, complication rates in young adults with type 1 diabetes (T1D) remain high and uptake of screening unclear. It is important to determine risk factors that restrain newly diagnosed patients from attending the health care services.

Methods: Our incident cohort included all 1832 patients who were diagnosed with T1D younger than 16 years of age between 1990 and 2009 at the Children's Hospital at Westmead, Sydney, NSW. Median age at diagnosis was 8.8 years [5.5-11.6] and 51% were female. Data was analysed from all their documented visits at our clinic between 1st January 1990 and 22nd February 2017 including personal data, clinical variables, blood tests. DCAS attenders (67%) were compared to non-DCAS attenders (34%) for: BMI, HbA1c and socioeconomic disadvantage (SES) which was based on postal codes of Australian Bureau of Statistics.

Results: Comparing non-DCAS attenders with DCAS attenders, diabetes clinic attendance in the first 2 years was 1.7 vs. 2.5 times per year (p<0.001), with a significant difference in their median HbA1c (7.9 vs. 8.0%, p=0.03) over this time, but no difference in BMI at 2-years. The difference in median lifetime HbA1c was even more significant (8.1 vs. 8.3%, p=<0.001). DCAS attenders continued in the Diabetes Service for a mean of 9.1 years whereas the non-DCAS attenders were followed for 3.8 years (p<0.001). 40% of the non-DCAS group were socio-economical disadvantaged compared to only 23% of the DCAS group (p<0.001). In multivariable analysis, DCAS attendance was associated with more visits in clinics over the first 2 years, less SES and higher median lifetime HbA1c.

Conclusions: SES and less frequent clinic attendance in the first 2 years after diagnosis but not obesity, were predictors of non-uptake of complication screening and are likely to influence the outcome of T1D for these patients. It remains unclear if the non-attenders had medical follow-up at other services.

Disclosure

J. Aulich: None. A.K. Chan: None. J. Cusumano: None. A. Pryke: None. P.Z. Benitez-Aguirre: None. M.E. Craig: None. K.C. Donaghue: None.
187-LB: Glucose Response Curve (GRC) Shapes in Mixed Meal Tolerance Tests (MMTTs) and Prediction of Response to Intervention in the TrialNet New-Onset Clinical Trials
Authors: NAJI ALMUTLAQ NOURAH , CLEVES MARIO A. , GITELMAN STEPHEN E. , DIMEGLIO LINDA , SOSENKO JAY M. , ISMAIL HEBA M.
Published: June 2019

It is known that individuals with a biphasic (BiP; two peaks) GRC during 2-hr oral glucose tolerance tests are at lower type 1 diabetes (T1D) risk than those with monophasic (MoP; one peak) and monotonic (MoT; continuous rise) GRCs. We hypothesized that GRC shapes during MMTTs in persons with newly-diagnosed T1D can predict response to intervention and that the shape may change in response to therapy. We performed an analysis to 1) Determine if the baseline GRC shape can predict response to intervention, and 2) Evaluate GRC changes in responders (R) vs. non-responders (NR) to interventions; response=AUC C-peptide (C-P) at 12 months not <5% of AUC C-P at baseline. Those with complete MMTT data at baseline (within 3 months of diagnosis) and at 12 months from all TrialNet intervention trials were included in the analyses (n=389, median (IQR) age 14.6 (8.1) years, BMI-z 0.6 (1.4), 56% male). At baseline, R had higher BiP GRCs [NR: 9.4% (25/227) vs. R: 20.6% (13/63); adjusted OR 2.92 (1.36-6.27); p=0.006)]. The overall shape distribution was significantly different between R and NR at 12 months (p=0.036) with a notable higher frequency of MoT GRCs in NR [NR: 30.1% (68/226) vs. R: 15.5% (9/58), p=0.031]. There were no differences between R and NR in the odds of changing from one GRC to another (p>0.05 for all).

In conclusion, the finding that a BiP GRC at baseline was more common in R than NR suggests that MMTT GRCs can be used to predict responses to interventions in those newly diagnosed with T1D. Further, the higher proportion of MoT GRCs in NR at 12 months suggests that MMTTs with MoT GRCs are indicative of metabolic decline.

Disclosure

N.N. Almutlaq: None. M.A. Cleves: None. S.E. Gitelman: Advisory Panel; Self; Avortes, Intermountain Therapeutics, Lilly Diabetes, Tolerion, Inc. Research Support; Self; Caladrius Biosciences, Inc., Janssen Research & Development. Other Relationship; Self; Novo Nordisk Inc. L. DiMeglio: Research Support; Self; Amgen Inc., Caladrius Biosciences, Inc., Janssen Research & Development, Medtronic, Sanofi. Other Relationship; Self; Dexcom, Inc. J.M. Sosenko: None. H.M. Ismail: None.

Funding

National Institutes of Health
1873-P: NAFLD and Insulin Resistance Is Associated with a Specific Signature in Membrane Phospholipid Metabolites in Adolescents with Polycystic Ovary Syndrome (PCOS)
Authors: CARREAU ANNE-MARIE , REYES YESENIA GARCIA , NEWCOMER BRADLEY , BEFROY DOUGLAS E. , BROWN MARK S. , CREE-GREEN MELANIE
Published: June 2019

Obese adolescents with PCOS are at high risk of nonalcoholic Fatty Liver Disease (NAFLD) which contributes to the risk of diabetes. Impaired hepatic energy metabolism is associated with diabetes and characterized by decreased inorganic phosphates (Pi) and ATP hepatic content. Membrane phospholipids measured by magnetic resonance spectroscopy (MRS) may reflect endoplasmic reticulum (ER) stress (phosphodiesters; PDE) or turnover/composition of membrane phospholipids: precursors (phosphomonoesters (PME): phosphoethanolamine (PE) and phosphocholine (PC)) and degradation products (PDE). It is not known if adolescents with PCOS and NAFLD have impaired hepatic energy metabolism or MRS signature associated with insulin sensitivity (Si). Obese adolescents with PCOS (n=25) underwent fasted hepatic 31phosphorus-MRS to measure ATP, Pi, PME and PDE relative to total phosphates (TP). Liver stiffness was measured with MR elastography and hepatic fat fraction (HFF) by MRI-proton density, with an HFF >5.5% classified as NAFLD. A 6h oral sugar tolerance test (OSTT, 25g fructose+75g glucose) was performed. Insulin sensitivity (Si) was calculated with the oral minimal model. 12 girls (BMI 36.4 ±7.3 kg/m2, age 16 (15-17)) had NAFLD and 13 did not (BMI 34.0±4.9 kg/m2, age 16 (14-17)). PME/TP were lower in girls with NAFLD (p=0.007) as well as the ratio of PME/PDE (p=0.002). A lower PE/TP was associated with higher liver stiffness (r= -0.55; p=0.009) and lower Si (r=0.57; p= 0.03). Total ATP/TP, y-ATP/TP and Pi/TP were similar between groups (p>0.15). Obese adolescents with PCOS and NAFLD have a distinct signature in hepatic 31P-MRS that is associated with Si suggesting alterations in membrane phospholipids. However, changes in energetics seen in adults with NAFLD are not present. Further work is needed to understand how these changes in hepatic phospholipids relates to insulin sensitivity and risk for diabetes.

Disclosure

A. Carreau: Advisory Panel; Self; Pfizer Inc. Y. Garcia Reyes: None. B. Newcomer: None. D.E. Befroy: None. M.S. Brown: None. M. Cree-Green: None.

Funding

National Institutes of Health (K23DK107871); Doris Duke Charitable Foundation; National Center for Advancing Translational Sciences (UL1TR002535); Diabetes Canada
1895-P: The Role of Endocannabinoid Receptor 1 (CB1R) in the Insulin Signaling Pathway
Authors: KIM YOO , EGAN JOSEPHINE M.
Published: June 2019

Cannabinoid 1 receptors (CB1Rs) are G protein-coupled receptors that are present in peripheral organs such as liver, pancreas, adipose tissue and skeletal muscle where they are involved in fine-tuning many metabolic functions. It was previously reported that liver-specific genetic deletion of CB1R (hCNR1-/-) mice fed high fat diets (HFD) had similar body weight to HFD-fed hCNR1+/+ mice, but they retained insulin sensitivity comparable to normal chow-fed (NCD) hCNR1+/+ mice. Therefore, this study was undertaken to uncover how hCB1Rs impact the insulin signaling pathway. Male hCNR1-/- and hCNR1+/+ mice were fed with a high fat high sugar diet (HFSD: N = 8-12) for 15 weeks. In contrast with the previous study of HFD only, HFSD-fed hCNR1-/- mice had less body weight gain than HFSD-fed hCNR1+/+ mice (25.1±2.7 vs. 19.8±1.7g), less fat accumulation in the liver, 1.3-fold increase in glucose disposal and 1.7-fold increase in insulin sensitivity compared to HFSD-fed hCNR1+/+ mice. Further study demonstrated that the lack of hepatic CB1R resulted in upregulated phosphorylation in liver of protein kinase B (AKT), causing activation of downstream target molecules, such as proline-rich AKT substrate 40 (PRAS40) in the mammalian target of rapamycin complex 1 (mTORC1). Similarly, primary hepatocytes isolated from hCNR1-/- mice had increased amounts of phosphorylated AKT and PRAS40 in comparison to hepatocytes from hCNR1+/+ mice. The co-immunoprecipitation studies clearly revealed hepatic CB1R modulates insulin signaling by the association or dissociation of the components of mTORC1. In support of these data, the deficiency of hepatic CB1R leads to upregulation of phosphoinositide 3-kinase (PI3K) class IB, p110γ, resulting in increased association between p110γ and G-protein beta. These findings indicate that genetic deletion of hepatic CB1R contributes to improved glucose homeostasis. Therefore, modulation of CB1R activity in liver may be a useful therapeutic in obese and diabetic individuals.

Disclosure

Y. Kim: None. J.M. Egan: None.
190-LB: Dipeptidyl Peptidase-IV Promotes the Developmental Programming of Chronic Inflammatory Diseases due to Perinatal Exposure to Maternal Obesity
Authors: RAMIL MONTANIEL KIM , BUCHER MATTHEW S. , MALOYAN ALINA
Published: June 2019

Perinatal exposure to maternal obesity promotes the development of chronic diseases such as cardiovascular (CVD) and metabolic diseases (T2DM) through a process known as developmental programming. The mechanisms of developmental programming are unclear. Dipeptidyl peptidase IV (DPP-4) is a pathogenic aminopeptidase that is upregulated in individuals with chronic diseases and its inhibitors are protective against T2DM and heart failure. DPP-4 can function as a membrane-anchored protein or as a soluble enzyme and can promote metainflammation and increased food intake. We found that DPP-4 enzymatic activity is upregulated over two-fold (P<0.05) in the cord blood of male offspring of women with obesity. Hence, we hypothesized that perinatal exposure to maternal obesity promotes the pathogenesis of cardiometabolic diseases through the activation of DPP-4. To test this hypothesis, we used a mouse model of maternal western style diet (WSD) feeding and found that offspring of WSD (Off-WSD) vs. regular diet (Off-RD) -fed mice have upregulated DPP-4 activity in plasma (two-fold, P<0.05) and PBMC (three-fold, P<0.05). This is concurrent with significantly impaired glucose tolerance, decreased insulin sensitivity, increased adiposity and hepatosteatosis. Systemic inhibition of DPP-4 with Sitagliptin alleviated these pathologies. Immune cells are a major source of plasma DPP-4 and we found that an allogeneic bone marrow transplantation (BMT) of Off-WSD bone marrow to lethally irradiated mice led to a significant increase in DPP-4 activity in plasma and PBMC compared to BMT with Off-RD bone marrow. This is concurrent with significantly increased adiposity, impaired glucose tolerance and decreased insulin sensitivity.

In summary, our findings suggest a pathogenic role of DPP-4, and specifically, immune-derived DPP-4, in the developmental programming of chronic inflammatory diseases due to perinatal exposure to maternal obesity.

Disclosure

K. Montaniel: None. M.S. Bucher: None. A. Maloyan: None.

Funding

National Institutes of Health; Oregon Health & Science University
1912-P: Portal Vein Metabolomic Profiling Highlights One-Carbon Metabolism as a Key Pathway Affected by Roux-en-Y Gastric Bypass Surgery
Authors: STEFATER MARGARET A. , AVILA JULIAN , CLISH CLARY B. , STYLOPOULOS NICHOLAS
Published: June 2019

After Roux-en-Y gastric bypass (RYGB) surgery, the intestine undergoes structural and metabolic reprogramming and appears to enhance utilization of energetic fuels including glucose and amino acids (AAs), changes that may be related to the surgery’s remarkable metabolic effects. Consistently, RYGB alters serum levels of AAs and other metabolites, changes hypothesized to underlie these benefits. To home in on the intestinal contribution, we performed metabolomic profiling in portal vein (PV) blood from lean, Long Evans rats after RYGB versus sham surgery. After RYGB, systemic blood (SB) metabolomic fingerprinting reflected changes in AA metabolism including serine/glycine/threonine metabolism, an important mechanism of one-carbon donation for folate metabolism. Principal component analysis (PCA) confirmed, among the metabolites most influential to the post-RYGB SB metabolome (PC1, P=0.0008), enrichment for these pathways as well as for sphingolipid and glycerophospholipid metabolism. PV profiling largely mirrored that of SB; using PCA to identify the PV metabolites most affected after RYGB (PC2, P=0.016), we found one-carbon metabolism (OCM) to be enriched. Among those metabolites significantly altered in PV versus SB, 15 were uniquely affected in sham-operated animals and 33 in RYGB-operated animals. Pathway analysis of the latter revealed overrepresentation of nitrogen, serine/glycine/threonine, and sphingolipid metabolism as well as ubiquinone biosynthesis. Together, our data provide novel insight into RYGB’s effects on the gut-liver axis and highlight a role for OCM as a key metabolic pathway affected by RYGB. OCM links cellular nutrient status to homeostatic mechanisms including purine biosynthesis, AA homeostasis, and epigenetic maintenance, and we hypothesize that in the intestine and liver, it might serve as a key fuel sensing pathway to mediate local changes in glucose metabolism, perhaps via epigenetic reprogramming.

Disclosure

M.A. Stefater: None. J. Avila: None. C.B. Clish: None. N. Stylopoulos: None.

Funding

National Institutes of Health
1913-P: Differences in Glucose Absorption in African American (AA) Patients Presenting with DKA and Severe Hyperglycemia (SH)
Authors: STEFANOVSKI DARKO , UMPIERREZ GUILLERMO E. , BOSTON RAYMOND C. , VELLANKI PRIYATHAMA
Published: June 2019

Approximately 70% of obese AA patients with new-onset diabetes presenting with DKA and SH have near-normoglycemia remission with intensive insulin treatment. In addition to improvements in insulin secretion and/or sensitivity as potential mechanisms of remission, we hypothesize that gut glucose absorption may be associated with remission. To estimate gut glucose absorption, we developed a novel mathematical model that simultaneously estimates SI and gut glucose half-life (GIGt1/2, inversely related to glucose absorption). This study aimed to compare insulin sensitivity estimates of the novel model to estimates from Dalla Man’s OGTT model and also assess GIGt1/2.

We included 42 AA patients who presented with new-onset DKA (n= 17) and SH (n=25). All subjects had a 75-gm 2-hour OGTT 1 week after insulin remission (REM) or at 3 months after continuous insulin therapy (no-REM). We estimated insulin sensitivity using our novel first-order absorption OGTT model (SInew) and GIGt1/2 and Dalla Man’s OGTT model (SI).

SInew was highly concordant with SI (R=0.61, P<0.001; Concordance=0.90). When separating subjects by DKA status, there were significant differences in the frequency of remission status (χ2=5.00, P=0.03). Subjects with DKA+REM (98±19 min) had a significantly longer GIGt1/2 compared to those with SH+no-REM (47±9 min, P=0.02). GIGt1/2 between DKA+no-REM (108±46 min) and SH+no-REM was also significantly different (P=0.045). SH+REM showed a statistical trend to be different from SH+no-REM (P=0.07). There were no significant differences identified between DKA+REM, DKA+no-REM and SH+REM.

The SInew from our novel OGTT model is highly concordant with SI from Dalla Man’s OGTT model. Our model additionally estimates GIGt1/2 and indicates slower gut glucose absorption is associated with remission. This new model shows that obese African American patients presenting with new-onset DKA or SH may benefit from interventions that target gut glucose absorption.

Disclosure

D. Stefanovski: None. G.E. Umpierrez: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Pharmaceuticals, Inc. Research Support; Self; AstraZeneca, Merck & Co., Inc., Novo Nordisk Inc., Sanofi US. R.C. Boston: None. P. Vellanki: Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Merck & Co., Inc. Research Support; Self; National Institutes of Health.

Funding

National Institutes of Health (K12HD085850 to P.V.)
1928-P: GDF15 Protects against Insulin Resistance in Individuals with Type 2 Diabetes
Authors: ZHANG HUI , DANTAS WAGNER , KING WILLIAM T. , AXELROD CHRISTOPHER L. , GIL SAULO , MURAI IGOR , ROSCHEL HAMILTON , GUALANO BRUNO , KIRWAN JOHN P.
Published: June 2019

Reduced insulin sensitivity is the putative defect in the onset of type 2 diabetes (T2D). Roux-en-Y gastric bypass (RYGB) can restore euglycemia and improve insulin sensitivity in individuals with T2D. However, the mechanisms contributing to surgical remission of T2D are not fully understood. Growth Differentiating Factor 15 (GDF15) is a circulating stress-responsive cytokine with appetite suppressive properties and therapeutic potential for treating obesity and T2D. However, the effect of GDF15 on insulin sensitivity is not known. The purpose of this investigation was to assess the role of GDF15 in insulin sensitivity in type 2 diabetic populations and to evaluate the effect of exogenous GDF15 treatment in obese diabetic mice. Twelve human volunteers with T2D (age: 40±2, BMI: 46.2±1.7) underwent RYGB surgery. Glucose tolerance and fasting plasma GDF15 levels were assessed prior to and 3-month post-surgery. Additionally, 8-week-old db/db mice were injected every other day for 4 weeks with saline or rhGDF15. After 4 weeks of treatment, body composition, insulin tolerance, and blood metabolites were assessed. Plasma GDF15 significantly increased after RYGB (PRE: 581.6±93.0 POST: 910.1±137.4 pg/ml, P<0.001). Change in GDF15 in humans with T2D correlated with improvements in insulin sensitivity based on the Matsuda index (R=0.6, P=0.05). Supporting our human data, 4 weeks of GDF15 treatment in db/db mice increased insulin sensitivity (P<0.05) while body weight (P<0.01), fasting blood glucose (P<0.05) and plasma insulin (P<0.05) were reduced.

In conclusion, GDF15 is associated with improved insulin sensitivity in humans with T2D, and increases insulin sensitivity in db/db mice. These data support a role for GDF15 as a therapeutic target for type 2 diabetes.

Disclosure

H. Zhang: None. W. Dantas: None. W.T. King: None. C.L. Axelrod: None. S. Gil: None. I. Murai: None. H. Roschel: None. B. Gualano: None. J.P. Kirwan: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases; National Institute on Aging; Louisiana Clinical & Translational Science Center