bluuebunny/crossref_metadata_2025_split huggingface dataset:
1818-P: Brainstem GABAergic Circuits Regulating Energy Balance
Authors: NECTOW ALEXANDER R. 
Published: June 2019

Obesity is a serious disorder for which there is currently no effective treatment. The hypothalamus has long been studied as the critical node for regulating food intake and body weight; however, no effective targets have yet been identified within the hypothalamus for anti-obesity therapy. There is thus a critical, unmet need to identify other loci within the central nervous system that: a) potently regulate energy balance, through alterations in food intake and energy expenditure, and b) are potential therapeutic targets. We have recently found that the brainstem’s dorsal raphe nucleus (DRN) houses a small population of inhibitory, GABAergic neurons (hereafter DRNVgat neurons), which potently regulates energy balance, and concomitantly body weight. This is accomplished through the direct regulation of both food intake and energy expenditure, the latter component of which is achieved through both behavioral and physiologic mechanisms. However, the circuit mechanism(s) through which these neurons regulate energy homeostasis are poorly understood.
In recent work, we made the unexpected finding that DRNVgat neurons project broadly throughout the brain to differentially regulate both food intake and energy expenditure. In particular, we have found that these neurons exhibit both ascending projections (to the bed nucleus of the stria terminalis, dorsomedial hypothalamus, and medial preoptic area), as well as descending projections to the raphe pallidus. This latter, descending projection forms part of a polysynaptic circuit anatomically linking DRNVgat neurons to brown fat. Using projection-specific optogenetic manipulations, we have found that these projections can all potently suppress thermogenesis, and differentially regulate food intake. Critically, these effects are scalable, with higher stimulation intensities leading to increasing effects on both feeding and thermogenesis. Together, this work uncovers a novel circuit mechanism through which DRNVgat neurons regulate energy balance.

Disclosure
A.R. Nectow: None.

Funding
American Diabetes Association/Pathway to Stop Diabetes (1-18-ACE-49, 1-18-IBS-148 to A.R.N.); Foundation for Prader-Willi Research; National Alliance for Research on Schizophrenia and Depression
1842-P: Enhanced TGF-ß1 Signaling in the Ducts of Cystic Fibrosis Pancreas Increases Basal Insulin Secretion by Islets
Authors: ROTTI PAVANA GURURAJ , APAK-EVANS IDIL , OMALLEY YUNXIA , NORRIS ANDREW , UC ALIYE , ENGELHARDT JOHN 
Published: June 2019

Cystic fibrosis (CF) is a multi-organ disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR protein is most highly expressed in pancreatic ductal epithelium, where loss of its function leads to ductal plugging, acinar destruction, and eventual diabetes. We hypothesize that exocrine pancreas remodeling in CF alters ductal paracrine signals to the endocrine pancreas. To test this hypothesis, we established primary ductal epithelium grown at an air-liquid interface (ALI) from WT and CFTR-KO ferret pancreas. Carbachol or IBMX/Forskolin (I/F) stimulated basolateral and apical secretions were analyzed for differentially secreted proteins using quantitative mass spectroscopy. Upon carbachol secretion 39 apical and 15 basolateral proteins were differentially secreted (P&lt;0.05) by CFTR-KO ducts. Similarly, 26 and 6 differentially secreted proteins (P&lt;0.05) were identified from I/F stimulated apical and basolateral secretions, respectively. TGF-β1 was one of the top upstream regulators of the differentially secreted proteins, P=5.02E-13 by Ingenuity Pathway Analysis. Single molecule fluorescence in situ hybridization of TGF-β1 mRNA on CFTR-KO and WT ferret pancreas indicated increased expression in CFTR-KO ducts compared to age matched controls. Likewise, expression of IGFBP7, a TGF-β1 signaling target, was augmented in fibroblasts surrounding pancreatic ducts and also in endocrine cells in CFTR-KO pancreas relative to controls. In cultured WT ferret islets, IGFBP7 treatment enhanced basal insulin secretion (2-fold, P&lt;0.05) and attenuated glucose stimulated insulin secretion. This IGFBP7-dependent insulin secretion phenotype was similar to that observed in cultured CFTR-KO islets, suggesting TGF-β1 signaling may be partly responsible for altered insulin secretion in CF.

Disclosure
P. Rotti: None. I. Apak-Evans: None. Y. OMalley: None. A. Norris: None. A. Uc: None. J. Engelhardt: None.

Funding
National Institutes of Health
1846-P: The Insulin-Sensitizing Effects of Palmitic Acid Esters of Hydroxy Stearic Acids Involve Intertissue Communication
Authors: ZHOU PENG , SANTORO ANNA , PERONI ODILE D. , NELSON ANDREW T. , SAGHATELIAN ALAN , SIEGEL DIONICIO , KAHN BARBARA 
Published: June 2019

Palmitic Acid esters of Hydroxy Stearic Acids (PAHSAs), bioactive lipids with anti-inflammatory and antidiabetic effects, improve glucose tolerance and insulin sensitivity in insulin-resistant mice. We found that PAHSA treatment enhances insulin action to suppress endogenous glucose production (EGP) in chow and HFD mice and promotes glucose uptake in glycolytic muscle and heart in HFD-fed mice. We aimed to determine the mechanisms by which PAHSAs enhance hepatic insulin sensitivity. We examined both direct effects in hepatocytes and indirect effects resulting from augmentation of insulin-mediated suppression of WAT lipolysis. Here, we show that PAHSAs inhibit lipolysis in WAT explants to the same extent as insulin. PAHSAs also enhance the anti-lipolytic effect of insulin in vivo, as indicated by a 30% reduction in FFAs during the clamp in PAHSA-treated mice under conditions with no suppression in Vehicle treated mice. To determine whether lowering FFAs is necessary for PAHSA effects on insulin sensitivity, we infused intralipid during a hyperinsulinemic-euglycemic clamp. In chow-fed mice infused with intralipid, PAHSAs still have a partial effect to lower EGP (Intralipid: vehicle 12±0.8 vs. PAHSA 7±1.7 mg/kg/minute), whereas in HFD-fed mice, preventing PAHSA-induced FFA reduction blocks PAHSA effects on hepatic insulin sensitivity (EGP Intralipid: vehicle 13±0.6 vs. PAHSA 12±1.7 mg/kg/minute). Our mechanistic studies show that PAHSAs inhibit basal EGP and reduce glucagon stimulated EGP directly in isolated hepatocytes. This is mediated by a cAMP-dependent pathway involving Gα/i protein-coupled receptors. Hepatic phospho-CREB is also reduced by PAHSAs in vivo. Sum: PAHSAs enhance hepatic insulin sensitivity through direct and indirect actions involving inter-tissue communication between adipose tissue and liver. These data reveal new mechanisms underlying the beneficial effects of PAHSAs, which will clarify their roles in healthy and disease states.

Disclosure
P. Zhou: None. A. Santoro: None. O.D. Peroni: None. A.T. Nelson: None. A. Saghatelian: None. D. Siegel: None. B. Kahn: Advisory Panel; Self; Alterna Therapeutics, Inc., American Diabetes Association, Harrington Discovery Institute, Janssen Research & Development. Consultant; Self; Ironwood Pharmaceuticals, Inc. Research Support; Self; National Institute of Diabetes and Digestive and Kidney Diseases.

Funding
American Diabetes Association (1-18-PDF-134 to P.Z.); National Institutes of Health (R01DK43051 to B.K.), (R01DK106210 to B.K., A.S.); JPB Foundation (to B.K.); American Heart Association (to A.T.N.)
1851-P: Bromocriptine (B) plus Curcumin (C) Interact to Reduce Advanced Fibrosis in a Mouse Methionine-Choline Deficient (MCD) Diet Model of Nonalcoholic Steatohepatitis (NASH)
Authors: TSAI TSUNG-HUANG , CINCOTTA ANTHONY 
Published: June 2019

NASH is characterized by severe hepatic fibrosis that potentiates cirrhosis and hepatocellular carcinoma. The MCD diet also potentiates severe liver fibrosis by reducing lipid secretion and antioxidant defenses, and adversely altering the gut microbiome to promote liver inflammation and damage. B, a dopamine D2 receptor agonist reduces adipose lipolysis and hepatic lipogenesis and inflammation. C improves a gut microbiome adverse to liver. C is also 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 against MCD diet induced liver fibrosis. C57B6 mice were fed an MCD diet for 8 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) daily for 4 weeks while maintained on the diet. A group of mice on normal diet were used as normal reference control. BC therapy led to significant reductions of serum ALT (85%), serum AST (33.4%), liver steatosis (65.4%), inflammation (86.7%), ballooning degeneration (60%), and NAFLD activity score (86.4%), as well as gene expression markers each of fibrosis (αSMA, TGFβ, COL1A, TXNIP) by 88-92%, inflammation (TNFa, IL1β, MCP1, iNOS) by 91-97%, ER stress (DDIT3, ATF, sXBP1, HSPA5) by 45-93%, steatosis (ACC1, PLIN2) by 31-97%, and fatty acid oxidation (PPARα, PGC1α, CPT1, UCP1) by 65-86, and reduced the percent fibrosis area by 92.7% relative to vehicle (and resultantly similar to normal diet mice) (all above: P&lt;0.05) and by amounts greater than B or C alone or additively. This study demonstrates the positive interactive effects of bromocriptine and curcumin on advanced liver fibrosis in an MCD diet model of NASH.

Disclosure
T. Tsai: Employee; Self; VeroScience LLC. A. Cincotta: Employee; Self; VeroScience LLC.
1858-P: Bidirectional Relationship between Chronic Kidney Disease and Nonalcoholic Fatty Liver Disease in Patients with Type 2 Diabetes Mellitus
Authors: HSU HUI-CHUN , CHANG YU HUNG , AN LINGWANG , LEE TAO-CHUN 
Published: June 2019

Background: Nonalcoholic fatty liver disease (NAFLD) and chronic kidney disease (CKD) are highly prevalent in type 2 diabetes mellitus (T2DM). Growing evidence suggests that NAFLD and CKD share common traditional risk factors, but it would seem unique risk pathways are involved as a consequence of NAFLD and CKD, efforts are still needed for a more comprehensive understanding of these relationship.
Methods: A structural equation model (SEM) was applied to assess associations between demographic data, inflammation factor, anthropometric and metabolic variables simultaneously and estimated glomerular filtration rate (eGFR) and fatty liver index (FLI). Given the potential that the NAFLD and CKD relationship may be bidirectional, SEMs also to examine direct and indirect effects of NAFLD on CKD, and vice versa.
Results: A total of 1,992 subjects with T2DM were enrolled in this study. Using multivariate analysis, NAFLD was independently associated with the risk of CKD (adjusted OR=1.59, 95% CI=1.12-2.25, P=0.009). SEMs showed that age (-0.216), triglyceride (-0.129), uric acid (UA, -0.415), albumin (0.163), and HbA1c (-0.066) levels had direct effects on eGFR, and the final model could explain 30% of the variability in eGFR. Age (0.094), triglyceride (0.174), body mass index (BMI) (0.707), UA (0.092), white blood cell (WBC) count (0.070), serum glutamic pyruvic transaminase (0.132) levels, and smoking (0.066) status had direct effects on FLI, and the final model could explain 67% of the variability in FLI. The common risk factors contributed to both eGFR and FLI were age, triglyceride, and UA. Unique risk factors were albumin and HbA1c for eGFR, and BMI, WBC, SGPT, and smoking for FLI. SEMs analysis confirm the bidirectional cause-relationships between NAFLD and CKD.
Conclusions: Common and unique risk factors and bidirectional relationship exist between CKD and NAFLD in patients with T2DM.

Disclosure
H. Hsu: None. Y. Chang: None. L. An: None. T. Lee: None.
1861-P: Pioglitazone Modulates Hepatic Pyruvate Dehydrogenase and TCA Cycle Flux
Authors: SHANNON CHRIS , RAGAVAN MUKUNDAN , DEFRONZO RALPH A. , MERRITT MATTHEW , NORTON LUKE 
Published: June 2019

Background: The insulin sensitizing actions of the antidiabetic drug Pioglitazone (PIO) have classically been ascribed to the reversal of ectopic lipid accumulation. We recently demonstrated acute, direct metabolic effects of PIO in hepatocytes, linking the suppression of hepatic glucose production to an inhibition of mitochondrial pyruvate metabolism. As the relevance of this mechanism in vivo is unclear, we studied the effects of PIO on hepatic pyruvate metabolism in a mouse model of insulin resistance.
Methods: Eight-week-old male ob/ob mice were placed on a regular diet (10% kcal fat) with (n=10) or without (n=10) 300 mg/kg PIO for four weeks before study during euglycemic-hyperinsulinemic clamp (5 mU/kg min). Endogenous glucose production (EGP), hepatic glycogen synthesis and anabolic/oxidative pyruvate flux ratio (PC/PDH) were determined from [3-3H] glucose infusion and 13C NMR isotopomer analysis using [2,3-13C] pyruvate. Livers were freeze-clamped in situ for pyruvate dehydrogenase (PDH), metabolite and protein analysis.
Results: PIO treatment lowered EGP by 62% (4.6 ± 1.0 vs. 12.1 ± 1.7 mg/kg min; P&lt;0.01) and increased glycogen synthesis 10-fold (19.7 ± 4.8 vs. 1.9 ± 0.8 μ mol/kg liver min; P&lt;0.005), whereas PC/PDH remained unchanged. PIO also suppressed liver PDH activation (14.6 ± 3.4 vs. 29.6 ± 4.3 µmol/kg liver min; P&lt;0.05), as well as acetylcarnitine (140 ± 18 vs. 297 ± 40) and acetyl-CoA (17.7 ± 2.4 vs. 47.8 ± 8.8 µmol/kg liver) concentrations (P&lt;0.01), whilst liver PDK4 protein expression was increased 46% compared to controls (P&lt;0.05).
Conclusions: Inhibition of PDH activation by Pioglitazone curtails mitochondrial acetyl-CoA delivery, relieving allosteric stimulation of pyruvate carboxylase and gluconeogenesis. Parallel reductions in PDH flux and EGP reconcile an unchanged PC/PDH ratio, suggesting an overall slowing of TCA cycle flux with PIO. These novel data highlight the therapeutic relevance of hepatic mitochondrial metabolism in the treatment of type 2 diabetes and liver disease.

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

Funding
University of Florida’s Southeast Center for Integrated Metabolomics (U24DK097209); National Institutes of Health
1865-P: Dual Chemokine Receptor CCR2/CCR5 Antagonist Cenicriviroc Ameliorates Lipotoxicity-Induced Insulin Resistance and NASH in Mice
Authors: OTA TSUGUHITO , CHEN GUANLIANG , NAGASHIMADA MAYUMI 
Published: June 2019

Hepatic lipid accumulation drives innate immunity with recruitment of bone marrow-derived macrophages and activation of liver-resident Kupffer cells, leading to insulin resistance and nonalcoholic steatohepatitis (NASH) development. However, treatment options that target macrophage/Kupffer cells recruitment or activation to halt the insulin resistance and NASH remain limited. A C-C chemokine receptor 2 (CCR2) plays a central role in macrophage recruitment. In addition, we reported that a different C-C chemokine receptor, CCR5, promotes insulin resistance and hepatic steatosis by regulating macrophage M1/M2 status. Here, we investigated the effect of cenicriviroc (CVC), a dual CCR2 and CCR5 receptor antagonist in a lipotoxic model of NASH with hepatic fibrosis and insulin resistance. Eight-week-old C57BL/6 mice were fed a high-cholesterol, high-fat (CL) diet or a CL diet containing 0.015% CVC (CL+CVC) for 12 weeks. The liver histology, insulin sensitivity, and fibrogenesis were examined. The CVC administration decreased the increase in plasma ALT levels as well as liver triglyceride, cholesterol, and TBARS levels caused by the CL diet. In addition, CVC improved glucose intolerance and hyperinsulinemia in the CL group and augmented the insulin signal, assessed by IRβ and Akt phosphorylation in the liver. Flow cytometry analysis revealed that mice fed CL+CVC had 72% fewer CD11c+CD206- M1-like macrophages, but 94% more CD11c-CD206+M2-like macrophages than CL group, indicating that CVC caused an M2-dominant shift in macrophages/Kupffer cells in the liver. Furthermore, CVC attenuated hepatic fibrosis by repressing the activation of hepatic stellate cells (HSCs) and decreasing hydroxyproline content. Thus, CVC ameliorated lipotoxicity-induced insulin resistance and NASH with hepatic fibrosis by polarizing M2 macrophage and attenuating HSC activation.

Disclosure
T. Ota: None. G. Chen: None. M. Nagashimada: None.

Funding
Japan Ministry of Education, Culture, Sports, Science and Technology
1866-P: Peripancreatic Adipose Tissue Protects against Ectopic Fat Accumulation in the Liver
Authors: CHANCLÓN BELÉN , MICALLEF PETER , NORDBECK ELIN BANKE , WU YANLING , WILDER BJÖRN , KANERVA JULIA , RORSMAN PATRIK ,   INGRID WERNSTEDT ASTERHOLM
Published: June 2019

Peripancreatic adipose tissue (PAT) is an understudied visceral fat depot that surrounds pancreas and is in touch with spleen and liver. Recent previous studies suggest that obesity-associated PAT inflammation accelerates pancreatic neoplasia in KrasG12D mice and that PAT conditioned-medium from pre-diabetic rats (9 weeks on high-fat diet; HFD) stimulates insulin secretion, proliferation and apoptosis in INS-1 cells. In this study, we aimed to better characterize PAT functionality in the context of HFD-induced obesity. PAT is a very small depot (0,5 ±0,04 mg) with a relatively high number of small adipocytes in unchallenged mice on chow, and we found that PAT expansion is less pronounced at early stages of HFD-feeding compared to the expansion of inguinal (IWAT), gonadal (GWAT) and mesenteric adipose tissues (MWAT). After 8-weeks HFD feeding the expansion of PAT was however comparable to the other three fat depots, but still PAT was more protected against the deleterious effects of HFD on adipose tissue inflammation and insulin resistance as judged by gene expression analysis (adiponectin, insulin receptor and inflammation markers). Feeding after 16-weeks HFD reduced the glucose and lipid uptake in all fat depots however, it was less pronounced in PAT; both fatty acid and glucose oxidation rates were higher in PAT compared to IWAT, GWAT and MWAT. Moreover, we noted that PAT is the only fat depot where the tissue weight correlates positively with liver weight in HFD-induced obese mice, and surgical removal of PAT followed by 16-week-HFD-feeding was associated with increased hepatic triglyceride content and accumulation of more and larger lipid droplets. PAT did not regenerate during this 16-week HFD time course suggesting that its precursor cells reside within the depot. Taken all data together, we propose that PAT, although it is located in the visceral compartment, can be considered as a metabolically/immunologically healthy fat depot that protects against hepatic steatosis in advanced obesity.

Disclosure
B. Chanclón: None. P. Micallef: None. E. Banke Nordbeck: None. Y. Wu: None. B. Wilder: None. J. Kanerva: None. P. Rorsman: None. I. Wernstedt Asterholm: None.
187-OR: Gut Microbiota Regulate Pancreatic Growth, Exocrine Function, and Gut Hormones
Authors: ALTINDIS EMRAH , SOTO MARION , ORLIAGUET LUCIE , HUANG QIAN , CEDERQUIST CARLY , DIRICE ERCUMENT , FUJISAKA SHIHO , HU JIANG , KULKARNI ROHIT , KAHN C. RONALD 
Published: June 2019

Growing evidence suggests a link between alterations in gut microbiota, diabetes, and metabolic syndrome. We recently showed that treatment with antibiotics to alter gut microbiota could improve insulin signaling and glucose metabolism in obesity- and diabetes-prone C57BL/6J mice on high-fat diet (HFD). This occurred in part through changing metabolite composition in plasma and decreasing adipose tissue inflammation. We also showed that these changes could be induced in germ-free mice by transferring the gut microbiota. It is known that diabetes is associated with exocrine pancreatic dysfunction, and diseases of the exocrine pancreas can be associated with diabetes. To explore the role of gut microbiota on host pancreatic exocrine function and its crosstalk with the endocrine system, mice were given either normal chow or HFD, or HFD and treatment with vancomycin or metronidazole. As expected, HFD alone produced signs of metabolic syndrome but also increased total pancreas weight by ~20%, an amount that could not be explained by islet hyperplasia. Analysis of the fecal proteome of these mice identified 138 host proteins of which 32 changed with either HFD or antibiotic treatment. Notably, the most significantly changed proteins by diet and altered microbiome were pancreatic enzymes: amylase, lipase and elastase. Furthermore, the plasma levels of glucagon-like peptide-1 (GLP-1) and peptide YY (PYY) levels were decreased, while glucose-dependent insulinotropic peptide (GIP) was increased by the HFD. These alterations of pancreas growth, acinar cell hyperplasia, exocrine pancreas function, and hormone secretion were reversed by antibiotic treatment of the host. These changes could also be reproduced in germ free mice by transfer of gut microbiota. Together, these data indicate that gut microbiota can regulate pancreatic growth and both exocrine and endocrine function. Better understanding pancreas-gut microbiome crosstalk can help us develop probiotics to cure pancreas dysfunction in diabetes.

Disclosure
E. Altindis: None. M. Soto: Employee; Self; LNC Therapeutics. Other Relationship; Self; biomeostasis. L. Orliaguet: None. Q. Huang: None. C. Cederquist: None. E. Dirice: None. S. Fujisaka: None. J. Hu: None. R. Kulkarni: None. C. Kahn: Advisory Panel; Self; MedImmune. Board Member; Self; Kaleido Biosciences. Consultant; Self; AntriaBio, Inc., Cobalt Therapeutics, Flagship Pioneering. Research Support; Self; Alnylam.

Funding
National Institutes of Health
1875-P: Liver-Specific p38 MAPK Stimulates Hepatic Fat Accumulation and Reduces Insulin Sensitivity In Vivo
Authors: RIVERS SYDNEY L. , WANG HUIQI , LAM LORETTA , GIACCA ADRIA 
Published: June 2019

Insulin resistance and hepatic fat accumulation are features of metabolic syndrome, although the mechanisms remain unclear. The cellular-stress responsive kinase, p38 Mitogen Activated Protein Kinase (p38), has controversial roles in regulating glucose and lipid metabolism in vivo. Here we show that liver-specific p38 stimulates hepatic triglyceride accumulation and contributes to the development of high fat diet (HFD)-induced insulin resistance. Liver-specific p38 knockout mice (p38KO) and wild type controls (WT) were fed a HFD or control diet for one week. Glucose tolerance tests revealed that HFD-fed p38KO mice had improved glucose tolerance compared to HFD-fed WT mice. Moreover, hyperinsulinemic-euglycemic clamps were performed to assess insulin sensitivity. HFD-fed p38KO mice were protected from reduced whole-body insulin action seen in HFD-fed WT mice, having similar glucose infusion rates as WT mice fed a control diet. To assess hepatic triglyceride accumulation, liver sections were stained with Oil Red-O post diet treatment. HFD-fed WT mice had significantly greater hepatic triglycerides, represented as a percentage of total liver area, compared to HFD-fed p38KO mice. In accordance with reduced hepatic triglyceride accumulation, decreasing trends in triglyceride secretion rates and plasma ApoB protein expression were reported between HFD-fed p38KO and HFD-fed WT mice following Poloxamer-407 injections. Overall, our data suggest that genetic inhibition of hepatic p38 may provide benefit in controlling heightened hepatic fat accumulation and reduced insulin sensitivity observed in HFD models.

Disclosure
S.L. Rivers: None. H. Wang: None. L. Lam: None. A. Giacca: Research Support; Self; Jazz Pharmaceuticals.

Funding
Canadian Institutes of Health Research