bluuebunny/crossref_metadata_2025 huggingface dataset:
278-OR: Increased Cortisol Excretion in Morbid Obesity Is Associated with Higher Body Weight but More Favorable Glucose Metabolism Parameters
Authors: BRIX JOHANNA , KRZIZEK EVA-CHRISTINA , TURA ANDREA , PACINI GIOVANNI , LUDVIK BERNHARD
Published: June 2019

Cortisol levels might be increased in obesity leading to enhanced gluconeogenesis and inhibited glucose utilization. Subsequent release of catecholamines might additionally promote obesity. In morbid obesity, the association of cortisol excretion to metabolic parameters is not well characterized. We included 1249 patients with MO (76.8% females, mean BMI 45.6± 6.2 kg/m², mean age 40±12 years). Anthropometric data and cardiovascular risk factors were assessed and a glucose tolerance test (oGTT) for calculation of Insulin Resistance and Sensitivity indices was performed. All patients collected 24h hour urine specimens to assess cortisol excretion on two consecutive days. Regarding cortisol excretion, patients were divided into three tertiles (fist< 39.1 µg/24h, second between 39.1and 139.9 µg/24h and third >139.9 µg/24h. Patients in the highest tertile had a higher BMI (p= 0.015) and waist circumference (p= 0.029), however, exhibited a lower diastolic blood pressure (p=0.001), 1-hour (p=0.001) and 2-hour glucose (p= 0.003) as well as HbA1c (p=0.047) values. Area under curve (AUC) for glucose (p<0.001) as well as insulin (p= 0.018) were lower, and HOMA-beta was increased (p=0.006). We did not find any differences regarding OGIS or QUICKI. In a correlation analysis, cortisol levels were inversely associated with glucose-AUC (r=-0.206; p<0.001), HOMA beta (r= 0.084; p= 0.005), BMI r= 0.099; p=0.002), glucose 1-hour (r=-0.107; p<0.001) and glucose 2-hour levels (r=-0.107; p<0.001). Despite being more obese, patients with higher cortisol excretion present with a more favorable metabolic profile compared to those with lower cortisol excretion. These results deserve further attention regarding the respective potential mechanisms.

Disclosure

J. Brix: None. E. Krzizek: None. A. Tura: None. G. Pacini: None. B. Ludvik: Advisory Panel; Self; Amgen Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Sanofi, Takeda Development Centre Europe Ltd. Research Support; Self; Akebia Therapeutics, Eli Lilly and Company, Novartis AG, Novo Nordisk A/S. Speaker's Bureau; Self; Merck Sharp & Dohme Corp.
283-LB: Dissociating Insulin Signaling and SREBP1c Action from the Lipogenic Drive Seen in Human and Murine Hepatic Insulin Resistance
Authors: VATNER DANIEL F. , TER HORST KASPER W. , GOEDEKE LEIGH , ZHANG DONGYAN , SAMUEL VARMAN , SERLIE MIREILLE J. , SHULMAN GERALD I.
Published: June 2019

The ongoing debate regarding pathway selective hepatic insulin resistance (IR) focuses on potential insulin signaling branchpoints beyond which regulation of glucose metabolism is attenuated and regulation of de novo lipogenesis (DNL) remains intact. These models suggest that impaired insulin receptor kinase activity, proximal to any potential branchpoint, only plays a minor role in clinical hepatic IR. We compared insulin signaling in liver biopsies from human subjects with nonalcoholic fatty liver disease (NAFLD) and hepatic IR vs. weight matched controls, using glucose ingestion to drive insulin secretion and fructose ingestion as a low insulin comparator. Insulin signaling through phosphorylation of insulin receptor β, Akt, and GSK3β was suppressed in NAFLD (all P < 0.05). Insulin regulation of mTORC1 via PRAS40 phosphorylation was absent in NAFLD subjects; no differences were seen between controls and NAFLD subjects in AMPK dependent RAPTOR phosphorylation. Pathway selective IR implies SREBP1c is primarily responsible for DNL in the insulin resistant liver; we probed the transcriptional regulation of DNL by assessing the in vivo occupancy of lipogenic target gene promoters by SREBP1c and ChREBP using ChIP experiments in mice with diet-induced IR. Fasted-refed mice were refed with high fat diet (HFD) and 1% dextrose drinking water. Mice with hepatic IR (9 day HFD fed) or hepatic and peripheral IR (4 week HFD fed) were compared with more insulin sensitive 2 day HFD fed mice. SREBP1c was not bound to the Fasn promoter in insulin resistant livers, while ChREBP was bound to the Fasn promoter equally in all three groups of mice.

Conclusions: There is a proximal block in insulin signaling in humans with hepatic IR and NAFLD, dissociating previously measured changes in DNL rates from insulin signaling. SREBP does not mediate DNL in insulin resistant mice while ChREBP action can explain the maintenance of the transcriptional program of lipogenesis.

Disclosure

D.F. Vatner: None. K.W. ter Horst: None. L. Goedeke: None. D. Zhang: None. V. Samuel: None. M.J. Serlie: Advisory Panel; Self; Fresenius Medical Care. G.I. Shulman: Advisory Panel; Self; AstraZeneca, Janssen Research & Development, Merck & Co., Inc. Advisory Panel; Spouse/Partner; Merck & Co., Inc. Board Member; Self; Novo Nordisk A/S. Consultant; Self; Aegerion Pharmaceuticals, IMetabolic BioPharma Corporation, Longitude Capital, Nimbus Discovery, Inc., Staten Biotechnology B.V.

Funding

National Institutes of Health
291-OR: Exercise Modulates Glucose Metabolism in Part through Changes in the Gut Microbiome
Authors: BRANDAO BRUNA , LI MENGYAO E. , CAI WEIKANG , BATISTA THIAGO M. , HIRSHMAN MICHAEL F. , GOODYEAR LAURIE J. , KAHN C. RONALD
Published: June 2019

Exercise is one of the most prescribed interventions in the treatment/prevention of diabetes and metabolic disease. It is well established that different diets affect the distribution of gut microbiota. It has recently been shown that exercise may also modify the gut microbome and these modifications may differ depending on the obesity status of the individual. In this work we investigated whether metabolic changes triggered by exercise are transferable via cecum microbiota transplantationand how this would impact on the obesity response to high fat diet (HFD). Ten-week old, B6J male mice, on chow diet, were reared under either standard sedentary conditions or with access to a running wheel for 4 weeks. Under these conditions the latter mice ran an average of 6 Km per day. Mice were scarified, fecal DNA were sequenced and the cecum microbiota were transplanted into germ-free mice in the presence/absence of HFD. As expected, exercise improved insulin sensitivity and glucose tolerance in the donor mice. Shotgun metagenomic analysis of gut microbiota revealed that exercise was associated with decreased relative abundance of Parasutterella_excrementihominis and Bacteroides_thetaiotaomicron and increased Clostridium clostridioforme CAG:511 and Lactobacillus murinus ASF361. Functional composition of the bacterial metagenome indicated these changes resulted in an upregulation of microbial pathways of valine and methionine metabolism. More importantly, transplant of cecum microbiota from exercised donors to non-exercise donors improved running capacity in lean mice and improved glucose tolerance in mice on HFD. In both groups, this was associated with increased glucose uptake in skeletal muscle soleus and BAT.

In summary, exercise reshapes the gut microbiome to improve whole body glucose metabolism, and this effect can be transferred by microbiome transplant. This provides an new opportunity in the treatment of obesity-related insulin resistance.

Disclosure

B. Brandao: None. M.E. Li: None. W. Cai: None. T.M. Batista: None. M.F. Hirshman: Stock/Shareholder; Self; Abbott Laboratories, AbbVie Inc., Amgen Inc., Colgate-Palmolive Company, Medtronic. L.J. Goodyear: 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 Institute of Diabetes and Digestive and Kidney Diseases
294-LB: Transcriptome-Wide Signals Converge Upon Mitochondrial Function in Developmental Stages of Type 2 Diabetes in Human Skeletal Muscle
Authors: SZCZERBINSKI LUKASZ , NIEMIRA MAGDALENA , SZCZERBINSKI KAROL , PUCHTA URSZULA , SIEWIEC ELWIRA , CITKO ANNA , ZAPOLSKA JOANNA , TAYLOR MARK , LARSEN STEEN , GORSKA MARIA , KRETOWSKI ADAM
Published: June 2019

Skeletal muscle is a major contributor to overall glucose homeostasis in the body and together with the liver is chiefly responsible for the whole-body insulin sensitivity. However, few studies have investigated the transcriptional signature of insulin resistance in skeletal muscles, at different stages of T2D development. Thus, the aim of this study was to identify the transcriptional signature of dysregulated glucose metabolism in skeletal muscle in humans by profiling the transcriptome with next-generation sequencing (NGS). We selected 40 sedentary males, aged 40-65 years, with BMI 26-33 kg/m2, diagnosed into three groups: 18 normoglycemics, 14 prediabetics and 8 subjects with type 2 diabetes. mRNA was extracted from biopsied Vastus lateralis muscles and sequenced with Illumina HiSeq 4000.Comparing normoglycemic and diabetic subjects revealed 4404 significantly differentially expressed (DE) genes, which represents around 30% of all mapped genes in studied samples. For the comparison between type 2 diabetics and prediabetics, the number of DE genes was 4055 genes. Interestingly we found no significantly DE genes between normoglycemics and prediabetics. KEGG enrichment analysis revealed that DE pathways between diabetics vs. nondiabetics were enriched for pathways related to oxidative phosphorylation, thermogenesis, nonalcoholic fatty liver disease and neurological diseases, which share the same underlying mechanism: mitochondrial dysfunction.These results indicate that skeletal muscle contributes most to overt T2D, with a possible indication that the liver contributes most to early stages the disease development. Differences observed between type 2 diabetics and the two remaining groups, occurring primarily in oxidative phosphorylation and electron transport chain pathways suggests that mitochondrial dysfunction plays the predominant role in the pathogenesis of T2D.

Disclosure

L. Szczerbinski: None. M. Niemira: None. K. Szczerbinski: None. U. Puchta: None. E. Siewiec: None. A. Citko: None. J. Zapolska: None. M. Taylor: None. S. Larsen: None. M. Gorska: None. A. Kretowski: None.
295-LB: Metformin Biphasic Effect on Human Cardiac Mitochondria
Authors: EMELYANOVA LARISA , BAI XIAOWEN , RUDIC THEODORE , WARNER CATHERINE , RIZVI FARHAN , ROSS GRACIOUS , KRESS DAVID , JAHANGIR ARSHAD
Published: June 2019

Mechanisms by which metformin (Mtf) protects human heart in type 2 diabetes remain debated. We hypothesize that Mtf has a biphasic effect on mitochondria: it causes activation of respiration at low doses via AMPK signaling and inhibition at high doses by directly affecting the activity of complex I and reducing oxidative stress. Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) and mitochondria isolated from human cardiac tissue were exposed to different doses of Mtf (0-20 mM) for 24h and acutely. Oxygen consumption rate (OCR) was assessed in the cells and mitochondria energized by glutamate and malate (complex I substrates) using XF96 Extracellular Flux Analyzer. Superoxide production was measured by monitoring changes in fluorescence intensity of dihydroethidium (DHE). At low doses (0.1 - 1 mM), Mtf significantly increased OCR in hiPSC-CMs (Fig. 1A). The highest OCR was observed at 0.5 mM Mtf. This effect was abrogated by compound C, an inhibitor of AMP-kinase (Fig. 1A). Mtf did not increase OCR in isolated mitochondria (Fig. 1B). At higher doses (1-20 mM), it inhibited OCR both in the cells and isolated mitochondria. This was associated with reduction of superoxide production in the mitochondria (Fig. 1C). In the human heart, Mtf improves cardioprotection due to its biphasic effect on mitochondria. This can be a beneficial alternative to patients with impaired endogenous cardioprotective responses.

Disclosure

L. Emelyanova: None. X. Bai: None. T. Rudic: None. C. Warner: None. F. Rizvi: None. G. Ross: None. D. Kress: None. A. Jahangir: None.

Funding

Aurora Health Care (N-570-3657)
298-OR: Sustained Improvement in Glycaemia following Flash Glucose Monitoring: An Expanded Worldwide Analysis
Authors: JANGAM SUJIT , LANG JORDAN , DUNN TIMOTHY , XU YONGJIN , HAYTER GARY
Published: June 2019

Flash glucose monitoring (FreeStyle LibreTM system) has shown sustained significant improvement in glycaemia over several months in clinical trials and the real world. Our aim was to re-evaluate the system in a longitudinal observational study with an expanded dataset. De-identified glucose data from 30,703 users over 6 months were analysed. The population was divided into tertiles of low, medium and high risk using time in hyperglycaemia (>240 mg/dl) and time in hypoglycaemia (<70 mg/dl) during the first two weeks. Each group was further sub-divided into tertiles of lower, medium and higher scanning frequency. Comparing the first and last bi-weekly periods, individuals in high risk tertile for hyperglycaemia showed reduced time >240 mg/dl (mean±SE) from 8.2±0.1 to 6.2±0.1 h/day in higher frequency scanners (p<0.0001) and 9.4±0.1 to 7.7±0.1h/day in lower frequency scanners (p<0.0001). Higher and lower frequency scanners in high risk tertile for hypoglycaemia showed reduced time <70 mg/dl (169±2 to 127±2min; p<0.0001 and 174±2 to 125±2min; p<0.0001). Time≤54mg/dL was also reduced in higher and lower frequency scanners (70±1 to 51±1min and 82±1 to 59±1min; p<0.0001 for both). Using an expanded dataset, we have confirmed that Freestyle Libre system use in real life settings is associated with sustained improvement in time spent in hyperglycaemia, hypoglycaemia and serious hypoglycaemia.

Disclosure

S. Jangam: Employee; Self; Abbott Laboratories. Employee; Spouse/Partner; Abbott Laboratories. J. Lang: Employee; Self; Abbott. T. Dunn: Employee; Self; Abbott Laboratories. Y. Xu: Employee; Self; Abbott Laboratories. G. Hayter: Employee; Self; Abbott.

Funding

Abbott Diabetes Care
3-OR: Differential Effects of Novel Antidiabetic Agents on Vascular Function Indices in Patients with Type 2 Diabetes Mellitus
Authors: SIASOS GERASIMOS , BLETSA EVANTHIA , STAMPOULOGLOU PANAGIOTA K. , BATZIAS KONSTANTINOS , PASCHOU STAVROULA A. , ANTONOPOULOS ALEXIOS , TSIGKOU VASILIKI , GOULIOPOULOS NIKOLAOS , MAZARIS SAVVAS , OIKONOMOU EVANGELOS , THANOPOULOU ANASTASIA , POLITOU MARIANNA , VRYONIDOU ANDROMACHI , TOUSOULIS DIMITRIS , TENTOLOURIS NIKOLAS
Published: June 2019

Background: Arterial stiffness is a well-established surrogate marker of vascular properties in patients with type 2 diabetes mellitus (DM2). We aimed to investigate whether optimization of DM2 therapy with novel antiglycemic agents additional to metformin, may improve arterial wall properties.

Methods: We enrolled 99 consecutive patients (male gender=63.3%) receiving metformin and still not achieving the therapeutic targets. Subjects were assigned to age and sex matched equal groups (n=33/group) of an additional antiglycemic agent; either DPP-4i, SGLT2i (n=28) or GLP-1 agonists. Applanation tonometry was used to assess non-invasively augmentation index (AIx) and aortic pulse wave velocity (PWV) as a measure of arterial stiffness at baseline and at 3-month follow-up. Among other demographics data, hemoglobin A1c (HbA1c) was measured.

Results: There was no difference for male gender (p=0.10) or age (64.92 ± 8.30 years, p=0.27) between the 3 study groups. Interestingly, baseline values improved significantly after SGLT2i and DPP-4i administration both for PWV (11.46 ± 2.77 vs. 9.83 ± 2.19 m/s and 10.89 ± 2.35 vs. 9.68 ± 1.77 m/s respectively, p=0.01 for both) and AIx (28.81 ± 8.55 vs. 25.82 ± 7.40 and 27.91 ± 13.05 vs. 24.91 ± 12.70 respectively, p=0.01 for both), when compared to those at follow-up time. In contrast, GLP-1A administration decreased PWV (12.82 ± 3.00 m/s at baseline vs. 11.67 ± 2.77 m/s during follow-up, p<0.001) but not AIx (31.64 ± 6.21 vs. 30.18 ± 6.03, p= 0.18). HbA1c at baseline was uniformly decreased in all study groups when compared to follow-up (7.52% vs. 6.72% for SGLT2i, 7.76% vs. 6.92% for DPP-4i and 8.19% vs. 6.85% for GLP-1A, p<0.001 for all).

Conclusion: The optimization of DM2 treatment with SGLT2i, DPP-4i or GLP-1A, added to metformin, not only helps to achieve better glycemic control but significantly ameliorates arterial stiffness indices and achieves therapeutic targets in patients with DM2.

Disclosure

G. Siasos: None. E. Bletsa: None. P.K. Stampouloglou: None. K. Batzias: None. S.A. Paschou: None. A. Antonopoulos: None. V. Tsigkou: None. N. Gouliopoulos: None. S. Mazaris: None. E. Oikonomou: None. A. Thanopoulou: None. M. Politou: None. A. Vryonidou: None. D. Tousoulis: None. N. Tentolouris: None.
302-LB: Mild Mitochondrial Stress in POMC Neurons Promotes Thermogenesis in Inguinal White Adipose Tissues and Prevents Diet-Induced Obesity
Authors: KIM SEONG JUN , MIN SE HEE , KANG GIL MYOUNG , KIM JI YE , CHOI JONG HAN , KIM MIN-SEON
Published: June 2019

Mild mitochondrial stress in POMC neurons promotes thermogenesis in inguinal white adipose tissues and prevents diet-induced obesity.

Normal mitochondrial dynamics and functions in hypothalamic neurons are pivotal in the maintenance of systemic energy homeostasis. CRIF1 has been identified as a mito-ribosomal protein which critically mediates incorporation of mitochondrial DNA (mtDNA)-encoded oxidative phosphorylation (OXPHOS) polypeptides into the inner mitochondrial membrane. Complete loss of CRIF1 in neurons causes severe OXPHOS dysfunction and leads to a neuronal death. To study the impact of mitochondrial stress in hypothalamic proopiomelanocortin (POMC) neurons, we generated a mice model lacking CRIF1 expression specifically in the POMC neurons using cre-lox system. Both male and female POMC-cre; CRIF1f/f mice developed obesity since the 10-15 weeks of age due to a gradual loss of POMC neurons. Unexpectedly, energy expenditure and thermogenesis were enhanced in POMC-cre; CRIF1+/f mice compared to their wild littermates. Moreover, they were resistance to diet-induced obesity when exposed to a high fat diet (HFD) before the 10 weeks of age. For the mechanism of enhanced thermogenesis, we found increased UCP1 expression and brown adipocyte-like cells in the inguinal subcutaneous white adipose tissue (ingWAT), so called "browning of subcutaneous fat depot." More strikingly, the expression of mitochondrial unfolded protein response (UPRmt) markers was significantly increased in the ingWAT of POMC-cre; CRIF1+/f mice. These findings strongly suggest that mild mitochondrial stress in POMC neurons causes mitochondrial stress response in the remote organ (ingWAT) via unidentified cell-non-autonomous mechanisms, leading to increased thermogenic activity in this fat depot.

Disclosure

S. Kim: None. S. Min: None. G. Kang: None. J. Kim: None. J. Choi: None. M. Kim: None.

Funding

National Research Foundation of Korea (NRF-2013M3C7A1056024)
307-OR: Interaction of Diabetes Genetic Risk and Successful Lifestyle Modification in the Diabetes Prevention Program
Authors: RAGHAVAN SRIDHARAN , JABLONSKI KATHLEEN A. , DELAHANTY LINDA M. , MARUTHUR NISA , LEONG AARON , FRANKS PAUL W. , KNOWLER WILLIAM C. , FLOREZ JOSE C. , DABELEA DANA ,   DPP RESEARCH GROUP
Published: June 2019

Intensive lifestyle intervention (ILS) targeting diet, physical activity (PA), and weight loss is efficacious for diabetes mellitus (DM) prevention, but not all high-risk individuals benefit. We tested whether DM genetic risk helps identify persons for whom ILS had a greater influence on DM risk. We studied 823 individuals in the ILS arm of the Diabetes Prevention Program with genetic and lifestyle data who were free of DM at one year after study enrollment. We estimated the associations of a DM genetic risk score (GRS) based on 67 loci from a recent genome-wide association study and achievement of three ILS goals at one year (7% weight loss, 150 minutes/week of self-reported moderate to strenuous leisure-time PA, and a goal for self-reported daily total fat intake) with incident DM using multivariable proportional hazards regression, including multiplicative interaction terms (i.e., on rate ratios) between GRS and each ILS goal. We used multivariable Poisson regression to estimate absolute risk reduction (ARR) in DM from ILS goal achievement across tertiles of the GRS, including tests for additive interaction (i.e., on rate differences) between GRS and each ILS goal. The GRS, and achieving each ILS goal and achieving all three ILS goals were associated with incident DM (all p<0.05; mean follow-up 3.0 years). There were no significant multiplicative interactions between GRS and achievement of any or all ILS goals. There were significant additive interactions between GRS and achievement of the weight loss (p<0.001), PA (p=0.03), and all three ILS goals (p<0.001). The ARR in incident DM associated with achievement of all three ILS goals increased across GRS tertiles: 1.8 [0.3, 3.4], 3.1 [1.5, 4.7], 3.9 [1.6, 6.2] fewer DM cases/100-person-years from the 1st to 3rd tertile. Genetic risk may identify high-risk subgroups for whom successful lifestyle modification is associated with greater absolute risk reduction of DM and a lower number needed to treat with lifestyle interventions.

Disclosure

S. Raghavan: None. K.A. Jablonski: None. L.M. Delahanty: Advisory Panel; Self; Jana Care Inc., Omada Health, Inc., Weight Watchers International, Inc. N. Maruthur: Other Relationship; Self; Johns Hopkins HealthCare Solutions. A. Leong: None. P.W. Franks: Board Member; Self; Zoe Ltd. Research Support; Self; AstraZeneca, Boehringer Ingelheim International GmbH, Lilly Diabetes, Novo Nordisk A/S, Novo Nordisk Foundation, Sanofi, Servier. W.C. Knowler: None. J.C. Florez: None. D. Dabelea: None. DPP Research Group: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases
31-OR: Skeletal Muscle Drp1 Maintains Metabolic Homeostasis through Preserving Mitochondrial Complex II Function
Authors: ZHOU ZHENQI , HEVENER ANDREA L.
Published: June 2019

Dynamin-related protein 1 (Drp1) is a primary mediator of mitochondrial fission, the process of dividing an individual mitochondrion into two organelles. We have previously shown that Drp1 expression is reduced in muscle of dysglycemic men compared to healthy normoglycemic men at rest and during exercise. Moreover, our laboratory has observed mitochondrial fission incompetence in several mouse models of skeletal muscle insulin resistance. To determine whether mitochondrial fission incompetence alters muscle metabolism and insulin action, we generated two mouse models with muscle-selective conventional and inducible deletion alleles of Drp1 (mDrp1KO and miDrp1KO respectively). Muscle deletion of Drp1 reduced fatty acid oxidation, promoted the accumulation of myocellular lipids, elevated the respiratory exchange ratio (RER) assessed by indirect calorimetry, and impaired insulin action. Examination of mitochondria from mDrp1KOmuscle using transmission electron microscopy revealed a notably elongated morphology compared to Controlf/f. Moreover, real time respirometry revealed an impairment in mitochondrial complex II activity that was a likely consequence of defective SDH assembly. Complex II assembly factor SDHAF2, required for flavination of the SDHA subunit and complex II activity, was markedly reduced in both Drp1 deletion mouse models and in Drp1 knockdown (KD) myotubes. Using immunoprecipitation approaches in mouse muscle and murine myotubes, we determined that SDHAF2 associates with Drp1, and that this relationship is critical for the translocation of SDHAF2 protein into mitochondria during fission. Pharmacological inhibition of complex II recapitulated the impairment in lipid oxidation and insulin action seen in mDrp1KO and Drp1KD cells. Our findings indicate that Drp1 is critical for fatty acid oxidation and the maintenance of insulin action by controlling the assembly and function of mitochondrial complex II.

Disclosure

Z. Zhou: None. A.L. Hevener: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (R01DK109724)
310-LB: Intracellular Protein Therapy with Truncated SOCS3 Suppresses Obesity
Authors: LEE KUYSOOK
Published: June 2019

Aim: Therapeutic molecule systemic delivery technology (TSDT), a platform for the discovery/development of new medicinal drugs, is enabled with a new generation of hydrophobic cell-penetrating peptides (CPPs), namely advanced macromolecule transduction domains (aMTDs). Leptin is an adipocyte-derived hormone that suppresses food intake and stimulates energy expenditure, leading to a reduction in stored body fat. However, patients with obesity have high levels of leptin, resulting in expression of suppressor of cytokine signaling (SOCS) 3 which is a negative feedback regulator of janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling initiated by the leptin receptor (ObR). Leptin resistance arises in part from the inhibition of leptin-initiated JAK/STAT signaling by SOCS3 binding to the phosphorylated Tyr-985 of ObR. We developed a cell-permeable, trans-dominant SOCS3 protein (CP-?SOCS3) to antagonize endogenous SOCS3 binding at P-Tyr-985 of ObR for use as an anti-obesity agent.

Method: We have developed CP-?SOCS3 by fusing the SOCS3 SH2-domain (46∼177 AA) to an aMTD sequence and tested the protein for the ability to enhance leptin induced ObR signaling, prolong leptin sensitivity and suppress weight gain in murine obesity models.

Result: CP-?SOCS3 interacted with ObR and enhanced leptin signaling (activation of STAT3) in mouse hypothalamus cells (mHypoA2/21 cells). In addition, CP-?SOCS3 showed 14.9% body weight loss in 14 days (15 mg/kg, daily, IV) in high fat diet-induced obese (DIO) mice. Food intake (33.6%), epididymal fat mass (34.1%) and serum leptin (59.5%) also decreased as compared to the mice treated with diluent. Furthermore, CP-?SOCS3 positively regulated leptin-initiated JAK/STAT signaling in DIO mice.

Conclusion: These results provide further validation of TSDT as a platform to develop protein-based biotherapeutics and suggest that CP-?SOCS3 has potential as a protein-based anti-obesity agent.

Disclosure

K. Lee: None.
313-LB: Weighting Approaches for a Genetic Risk Score and an Oxidative Stress Score for Predicting the Incidence of Obesity in a Korean Population
Authors: KIM MINJOO , JIN YOO HYE , HAN YOUNGMIN , HO LEE JONG
Published: June 2019

Background/Objectives: We aimed to predict the incidence of obesity in a Korean population using a genetic risk score (GRS) constructed with obesity-related single nucleotide polymorphisms (SNPs) along with an oxidative stress score (OSS).

Subjects/Methods: A total of 9,460 Korean subjects and 356,974 SNPs were included. The GRS was constructed using three significant obesity-related SNP loci, and the OSS was calculated with three reliable oxidative stress biomarkers.

Results: In an association study, the top three SNPs, rs2846412, rs58424360, and rs591120, showed profound associations with obesity in a Korean population (all FDR < 0.1). The GRS showed a more significant association with increased obesity (OR = 2.879, 95% CI = 2.283-3.630) than individual SNPs did after adjusting for age and sex. Three oxidative stress biomarkers, including malondialdehyde (MDA), oxidized low-density lipoprotein (ox-LDL), and 8-epi-prostaglandin F2α (8-epi-PGF2α), showed significantly high levels in the obese group. Moreover, the OSS, which was the sum of each oxidative stress biomarker score, showed a markedly high association with the incidence of obesity, with an OR of 3.213 (p = 3.57E-18). Based on the results from the regression tests and a receiver-operating characteristic (ROC) curve analysis, we found that HOMA-IR, hs-CRP, the GRS, and the OSS were the most relevant factors for the incremental risk of obesity and were significantly associated with the incidence of obesity. The area under the ROC curve (AUROC) was improved when the GRS was added to the model with the OSS and two obesity-related parameters (from 74.2% to 75.1%).

Conclusions: We first identified that subjects with an obesity GRS and a high OSS may have a higher risk of obesity. Our findings and weighting approaches were effective in predicting the incidence of obesity; furthermore, the GRS is a relevant factor that significantly predicts the risk of obesity.

Disclosure

M. Kim: None. H. Yoo: None. Y. Han: None. J. Lee: None.

Funding

Korean Ministry of Health and Welfare (HI13C0715); Korean National Research Foundation (NRF-2017R1C1B2007195, NRF-2012M3A9C4048762)
313-OR: PACAP from the Ventromedial Hypothalamic Nucleus Controls Both Energy Balance and Glucose Homeostasis
Authors: BOZADJIEVA NADEJDA , ROSS RACHEL A. , LOWELL BRADFORD , FLAK JONATHAN N.
Published: June 2019

Obesity results from energy imbalance, due, at least in part, to suppressed energy expenditure. While the brain tunes energy expenditure, the neurocircuits, and essential components within these circuits, that can accelerate and decelerate energy consumption have not yet been revealed in great detail. The ventromedial hypothalamic nucleus (VMN) is a critical site for the control of sympathetic tone, which controls a range of physiologic indices including energy expenditure. While the VMN contains, almost exclusively, glutamatergic neurons, neuropeptides are also expressed within the nucleus. Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) is the most abundant neuropeptide in the VMN. Since previous studies have implicated PACAPVMN in responses to energy status, we hypothesized that PACAPVMN is necessary for energy balance by promoting energy expenditure. We acquired a PACAPflox mice that, in the presence of cre, results in a truncated protein, preventing communication downstream by PACAP. Because PACAP is expressed in the periphery, we administered a VMN injection of AAVcre in PACAPflox mice. Bilateral AAVcre administration ablated dmVMN PACAP expression. We found that, in both male and female mice, loss of PACAPVMN results in massive obesity, adiposity, and hyperglycemia. Surprisingly, food intake only increased once body weight had almost doubled. In support of this notion, PACAPVMNKO suppressed VO2 and not activity, despite massive obesity. In addition to energy imbalance, PACAPVMNKO induced hyperinsulinemia and suppressed glucose clearance to brown adipose tissue, heart, and muscle during hyperinsulinemic/euglycemic clamp. Our data demonstrate that PACAP is an essential component for the promotion of energy expenditure and glucose clearance by the VMN. Future studies will reveal both the VMN cells that use PACAP for these functions, as well as the downstream cells that control energy balance and glucose homeostasis.

Disclosure

N. Bozadjieva: None. R.A. Ross: None. B. Lowell: None. J.N. Flak: Research Support; Self; Novo Nordisk Inc.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-17-INI-15 to J.N.F.)
314-LB: Dapagliflozin Induced Weight Loss and Metabolic Effects in Conjunction with a Low Carbohydrate Diet
Authors: HANSON PETRA , RANDEVA HARPAL , BARBER THOMAS M. , OHARE JOSEPH P.
Published: June 2019

Type 2 diabetes mellitus (T2DM) is closely associated with obesity. Lifestyle modification, including dietary modification, leads to weight loss, a crucial part of management of patients with T2DM. Evidence shows that treatment with Dapagliflozin leads to weight loss. However, there is a lack of in-depth studies exploring the body composition changes induced by Dapagliflozin in patients following low carbohydrate diet. Our aim was to undertake a detailed assessment of the appetitive and metabolic effects of Dapagliflozin in patients with T2DM. Participants had detailed metabolic studies, including indirect calorimetry (energy expenditure measurements), appetite assessment and quantification of body fat using BodPod in the Human Metabolism Research Unit; University Hospitals of Coventry and Warwickshire (UHCW), UK. The study duration is for 12 months; interestingly, preliminary data from 10 participants who completed 6 months follow-up showed statistically significant weight loss of 5.7 kg at month 6 (Figure 1). There was significant decrease in fat mass observed at month 3 (6.3kg) with no difference in muscle mass. There was no difference in energy expenditure before and after 12 months treatment with dapagliflozin. There was no change in self-reported appetite. These results indicate that low carbohydrate diet improves Dapagliflozin's potential for weight loss, with fat loss in particular.

Disclosure

P. Hanson: Other Relationship; Self; AstraZeneca, AstraZeneca. H. Randeva: None. T.M. Barber: None. J.P. OHare: None.

Funding

AstraZeneca; University Hospital of Coventry and Warwickshire
315-OR: Two Brown Adipocyte Subpopulations with Distinct Thermogenic Activity Coexist and Undergo Dynamic Interconversions upon Changes in Environmental Temperature
Authors: SONG ANYING , JANG MIN JEE , WU XIWEI , JIANG LEI , SCHERER PHILIPP E. , WANG QIONG
Published: June 2019

Brown adipose tissue (BAT) exerts beneficial metabolic effects on obesity and insulin resistance. Unmasking the complex physiology of BAT thermogenesis is essential to improve our ability to identify effective therapeutic approaches for metabolic disorders. Utilizing the AdipoChaser mouse, we discovered that there are two subpopulations of “classical” brown adipocytes, adiponectin “low expressers” and “high expressers,” co-existing within the BAT. Single-cell sequencing confirms that there are two brown adipocyte subpopulations that differ by adiponectin expression. Low adiponectin expressers also have lower UCP1 expression. We refer to the adiponectin low expresser as low thermogenic brown adipocyte (BA-L), and adiponectin high expresser as high thermogenic brown adipocyte (BA-H). The BA-L has larger lipid droplets, lower mitochondrial content, and higher mitochondrial membrane potential, reflecting reduced thermogenic activity. The BA-H subpopulation is enriched of genes involved in lipolysis, fatty acid oxidation, glycolysis, and oxidative phosphorylation; while the BA-L subpopulation is enriched of genes involved in fatty acid uptake and cell-to-cell trafficking. Upon changes in environmental temperature, the two brown adipocyte subpopulations undergo dynamic inter-conversions. Cold exposure converts BA-L into BA-H, while a thermoneutral environment does the opposite. Developmentally, brown adipocytes initiate differentiation between embryonic day 10 and 16 and they uniformly develop into BA-H. The BA-L subpopulation emerges postnatally. Importantly, the conversion of BA-L into BA-H upon cold exposure is impaired by age, but not by high fat diet feeding. Our results reveal the functional heterogeneity of brown adipocytes. Rather than being uniformly regulated, the thermogenic activity of BAT is controlled by the number of brown adipocytes recruitable into the BA-H pool.

Disclosure

A. Song: None. M. Jang: None. X. Wu: None. L. Jiang: None. P.E. Scherer: None. Q. Wang: None.

Funding

American Diabetes Association (1-19-JDF-023 to Q.W.); National Institutes of Health (K01DK107788R03, HD095414); City of Hope
318-OR: Targeting PPARγ Phosphorylation Prevents Obesity by Inducing a Brown-Like Healthy Remodeling of Adipose Tissue
Authors: RABHI NABIL , FARMER STEPHEN R.
Published: June 2019

The prevalence of obesity—and thus the incidence of metabolic diseases, including type 2 diabetes and cardiovascular disease—is dramatically increasing worldwide. Despite the critical role of adipose tissues in nutrient metabolism, there are surprisingly few clinically-approved medications targeting adipose tissue. Thiazolidinediones (TZDs), PPARγ agonists are exceptions that act in adipose tissue, but they are associated with adverse effects such as weight gain, and heart failure. An exciting series of studies demonstrate that posttranslational modifications can selectively enhance the insulin sensitizing activity of PPARγ. Our laboratory previously showed that roscovitine (RS), a CDK inhibitor, prevents diet-induced obesity by blocking PPARγ S273 phosphorylation, which promotes formation of UCP1+ (brite) adipocytes in WAT. RNA profiling reveals that brite adipocytes are distinct from beige adipocytes, which arise through catecholamine signaling suggesting that brown-like adipocytes in WAT may arise from multiple origins. Using adipocytes and progenitor’s lineage tracing mice, we demonstrate that brite adipocytes originate from mature white adipocytes through trans-differentiation. Inhibition of S273 phosphorylation of PPARγ in obese mice prevents weight gain, restores insulin and glucose sensitivity and a normal pancreatic β-cells response to glucose as well as prevents liver steatosis and adipocytes formation in bone. Moreover, mass spectrometry analysis of extracellular matrix proteins showed a reduced adipose tissues fibrosis following RS treatment. Mechanistically, inhibition of PPARγ phosphorylation shifts its interaction with cofactors favoring a brown-like healthy remodeling of adipose tissue, which includes suppression of genes associated with inflammation and fibrosis. Overall, we demonstrate targeting PPARγ phosphorylation using RS can constitute a new therapy for obesity and its metabolic complications.

Disclosure

N. Rabhi: None. S.R. Farmer: None.

Funding

American Heart Association (17POST33660875); Francophone Society of Diabetes (R01DK102199-01)
323-LB: Activation of the GLP-1 Receptor Signaling Prevents Beta-Cell Dedifferentiation in a Mouse Model of Wolfram Syndrome through Modulating TXNIP Expression
Authors: TANABE KATSUYA , AMO-SHIINOKI KIKUKO , HATANAKA MASAYUKI , TANIZAWA YUKIO
Published: June 2019

Loss of functional β cells results in a gradual progression of insulin insufficiency in Wolfram syndrome caused by recessive WFS1 mutations. Currently, there are no specific treatment recommendations for the disease. We have so far evidenced that, in the Wfs1-/- mice, β cells become dedifferentiated and revert to endocrine progenitor-like cells expressing Ngn3, and a subset of β cells takes α cell fate. Such β cell plasticity appears after nursing, independently of hyperglycaemia, and becomes more apparent along with diabetes progression accompanied with no significant increase in apoptosis. In this study, we examined the potential effects of a GLP-1 receptor agonist on β cell plasticity and glucose homeostasis in young Wfs1−/− mice without noticeable hyperglycaemia.

Results: Four weeks administration of Ex-4 resulted in an off-drug amelioration of glucose excursions after glucose loading in Wfs1−/− mice, with insulin secretory dynamics that were indistinguishable from those in WT mice, despite no alteration in beta cell mass. In association with the functional improvements, Ex-4 alleviated β cell dedifferentiation evidenced by a decrease of MafA and a Ngn3 emergence. At the same time, Ex-4 reversed a robust increase in thioredoxin interacting protein (TXNIP) in Wfs1-/- β cells. Mechanistically, Ex-4 suppressed it at both transcriptional and post-translational levels. Furthermore, we have found that genetic ablation of Txnip in Wfs1-/- mice inhibited β cell dedifferentiation, resulting in prevention of diabetes progression with preservation of β cell mass. Our finding implies that activation of the GLP-1 receptor signaling may prevent β cells from becoming dedifferentiated by modulating Txnip and may slow diabetes progression in Wolfram syndrome.

Disclosure

K. Tanabe: None. K. Amo-Shiinoki: None. M. Hatanaka: None. Y. Tanizawa: None.

Funding

Japan Society for the Promotion of Science (16K09752, 15K21198, 15K09390); Japan Diabetes Society; Japan Association for Diabetes Education and Care
327-LB: Transgenic Overexpression of MicroRNA-30d Impairs Glucose Homeostasis by Regulating Beta-Cell Proliferation and Dedifferentiation
Authors: TANG XIAOQING
Published: June 2019

MicroRNAs (miRNAs) are small non-coding RNAs that bind to target mRNAs and inhibit the translation in mammals. Abnormal miRNA functions are closely associated with pancreatic beta-cell loss and dysfunction in type 2 diabetes. Downregulation of miR-30d has been observed in the islets of diabetic mice. To investigate the in vivo function of miR-30d in pancreatic beta cells, we generated the transgenic mice that overexpressed miR-30d specifically in the pancreatic beta cells. Compared to wild type mice, the transgenic mice that overexpressed miR-30d resulted in hyperglycemia and glucose intolerance with reduced beta-cell mass and insulin production following high fat diet treatment. Strikingly, loss of beta-cell mass was partially due to beta-cell dedifferentiation to glucagon-producing alpha-cells, resulted in increased plasma glucagon level. The transcriptomic analysis combined with bioinformatic analysis of islets isolated from transgenic verses control mice identified connective tissue growth factor (CTGF) and Cyclin E2 as miR-30d potential targets. Moreover, several β-cell disallowed genes (e.g., MCT4/SLC16A3, LDHA) were observed upregulated in transgenic mice, which have been reported as positive markers of dedifferentiated human β-cells from type 2 diabetic patients. Taken together, miR-30d plays an essential role in maintaining mature beta-cell identity.

Disclosure

X. Tang: None.

Funding

National Institutes of Health
330-OR: A 30% Increased Insulin Dose Is Necessary to Control Postprandial Hyperglycemia for High Protein Meals in Individuals with Type 1 Diabetes Using Insulin Pump Therapy
Authors: PATERSON MEGAN A. , SMART CARMEL E. , RAFFERTY JORDAN , FENTON BRETT , PRICE DARRELL , FOSKETT DEBORAH C. , MAHER ANDREW , HOWLEY PETER , KING BRUCE R.
Published: June 2019

Dietary protein increases the postprandial glycemic response and insulin requirements in people with type 1 diabetes (T1D). The aim of this study was to determine the appropriate amount of additional insulin required for the protein content of a meal to prevent postprandial hyperglycemia. Twenty-nine subjects (12 male) with T1D using CSII (HbA1c<8.1%/65mmol/mol) aged 10-40 years consumed 5 test meals containing 50g protein, 0g fat and 30g CHO. Insulin doses were calculated based on the individual insulin:CHO ratio with 0 (standard), 15, 30, 45 and 60% additional insulin added to the dose and delivered over 3 hours using a dual/combination bolus. Postprandial glycemia was assessed by 4 hours of CGM. Standard insulin dosing resulted in delayed and sustained postprandial hyperglycemia. From 150 minutes, additional insulin doses of 30% resulted in significantly lower postprandial glycemic excursions, returning to baseline by 4 hours; compared with 0-15% additional insulin (p<0.05). Additional insulin doses of 45 and 60% resulted in increased episodes of hypoglycemia in 50% (n=15) and 80% (n=23) of participants respectively. This study demonstrates that adding 30% more insulin to a standard meal-time dose for a high protein meal, delivered using a dual/combination bolus, improves postprandial glycemia in individuals with T1D using CSII.

Disclosure

M.A. Paterson: None. C.E. Smart: None. J. Rafferty: None. B. Fenton: None. D. Price: None. D.C. Foskett: None. A. Maher: None. P. Howley: None. B.R. King: None.

Funding

John Hunter Children's Hospital; Dexcom External Research Program
331-LB: A Switch in Nutrient Sensing Controls Functional Maturation of Pancreatic Beta Cells
Authors: HELMAN AHARON , MELTON DOUGLAS
Published: June 2019

A drastic transition at birth, from constant maternal nutrient supply in-utero to intermittent postnatal feeding, requires functional changes in the metabolic system. Considering their central role in controlling metabolic homeostasis, little is known about how beta-cells adjust to the new nutritional challenge. Our experiments show that after birth beta-cells undergo a functional shift from amino acids- to glucose-stimulated insulin secretion. Moreover, a change in mTORC1 activity, from dependence on amino acids in the amino acid-rich environment in-utero to a postnatal dependence on glucose, corresponds with the initiation of glucose-responsive insulin secretion. Disrupting nutrient sensitivity of mTORC1 in mature beta-cells, via deletion of leucine-responsive inhibitors Sestrin1 and Sestrin2, reverts beta-cells to an immature state. Finally, manipulating mTORC1 nutrient sensitivity in stem-cell-derived beta-cells in-vitro shifts them from constitutive to glucose-dependent insulin secretion. These results reveal a post-transcriptional mechanism where nutritional environment and mTORC1 signaling regulate glucose responsiveness, thereby promoting beta-cell maturation.

Disclosure

A. Helman: None.
334-OR: Supplementation with Histidine-Containing Dipeptides Improves Cardiometabolic Risk Factors: A Meta-analysis of Randomized Controlled Trials
Authors: MENON KIRTHI ARAVIND , MOUSA AYA , MARQUINA CLARA , DE COURTEN BARBORA
Published: June 2019

Background: The burden of obesity and type 2 diabetes is rapidly increasing. Carnosine (β alanyl L-histidine), a dipeptide with anti-inflammatory, anti-glycating, and anti-oxidative properties, may be a potential strategy to mitigate cardiometabolic risk factors. However, studies examining the effects of carnosine and related histidine-containing dipeptides (HCDs; e.g., beta-alanine) on cardiometabolic risk have been inconsistent.

Objective: We conducted a systematic review and meta-analysis examining the effects of all HCDs on cardiometabolic risk factors in all population groups.

Methods: MEDLINE, CINAHL, EMBASE, and EBM were searched for randomized controlled trials investigating the effects of carnosine and/or HCDs on cardiometabolic risk factors, compared with placebo, usual care, or other interventions. Meta-analyses using random-effects models were used to calculate the mean difference (MD) and 95% confidence interval (CI) for each outcome.

Results: Forty- nine RCTs totaling 1709 participants were included. Carnosine and/or HCD supplementation lowered waist circumference (n= 162, MD [95% CI]= -3.53 cm [-5.65,1.41], p=0.001); post-prandial glucose (PPG) (n=230, MD [95% CI]= -0.79 mmol/L [-1.23,-0.35], p=0.0004), HbA1c (n=283, MD [95% CI]= -0.37% [-0.64,-0.10], p=0.006) and C-reactive protein (CRP) (n=272, MD [95% CI]= -0.43 mg/L [-0.64,-0.22], p< 0.0001) compared with control. In sensitivity analyses of RCTs using carnosine only, PPG, HbA1c, and CRP remained significantly lower with carnosine supplementation (all p<0.05), and fasting glucose and triglycerides became significantly lower with carnosine versus control (n=309 for both, MD [95% CI]= -0.58 mmol/L [-1.12,-0.05], p=0.03; and -0.2 mmol/L [-0.27,-0.13], p<0.0001, respectively).

Conclusion: Our findings provide level 1 evidence that carnosine and HCDs have potential benefits for prevention and treatment of diabetes.

Disclosure

K. Aravind Menon: None. A. Mousa: None. C. Marquina: None. B. de Courten: None.