bluuebunny/crossref_metadata_2025 huggingface dataset:
1578-P: Hyaluronidase-1 Inhibits Hepatic Gluconeogenesis
Authors: CHEN XI , ZHU YI
Published: June 2023

Hyaluronidase-1 (Hyal1) is a lysosomal hyaluronidase that intracellularly hydrolyzes hyaluronan (HA). Studies have demonstrated the critical role of Hyal1 in the development of diabetes-induced endothelium dysfunction. However, whether it plays a role in the development of glucose intolerance and obesity remains unclear. We treated Hyal1 whole-body knockout (Hyal1 KO) mice and liver specific overexpression (OE) of Hyal1 mice with a low-fat diet (LFD) or high-fat diet (HFD), Hyal1 KO mice gained similar body weight but developed glucose intolerance compared to littermate wildtype (WT) mice on HFD, the latter is primarily contributed by an increase in gluconeogenesis in the Hyal1 KO liver. In contrast, Hepatic OE of Hyal1 suppressed gluconeogenesis. However, the change in gluconeogenesis is unlikely mediated by digestion of extracellular HA by Hyal1 in the liver as albumin-driven expression of a more potent hyaluronidase Ph20 had no effect on glucose tolerance nor gluconeogenesis from pyruvate. Liver metabolites in the gluconeogenesis pathway were significantly increased by HFD, whereas this was significantly attenuated in hepatic Hyal1-OE mice. Glucose transportersGLUT1 and GLUT2 were not affected by Hyal1 OE. Hyal1 deletion does not affect overall gene expression, as revealed by mRNA-sequencing of liver. However, ATP generation measured by OCR before and after inhibiting ATPase is decreased in hepatocytes OE of Hyal1, which likely contributes to the reduced energy-consuming gluconeogenesis process in Hyal1 overexpressing hepatocytes. In summary, Hyal1 regulates gluconeogenesis independent of changes in extracellular HA content, gene expression, or glucose transporter abundance in hepatocytes. Reduced gluconeogenesis is likely due to the reprogramming of intracellular metabolism and a reduction in ATP production through a remain-to-understand lysosome to mitochondria crosstalk.

Disclosure

X.Chen: None. Y.Zhu: None.

Funding

U.S. Department of Agriculture-Agricultural Research Service (3092-51000-062)
1579-P: Impact of Adipose Tissue Insulin Sensitivity on Hepatic Insulin Sensitivity Preservation in Longitudinal Observation of Patients with NAFLD
Authors: SHIGIYAMA FUMIKA , HIRUMA SHIGENORI , KUMASHIRO NAOKI
Published: June 2023

Hepatic insulin resistance associated with NAFLD is a major pathogenesis of type 2 diabetes and its complications. In addition to reducing lipids in the liver, identifying the factors that may preserve hepatic insulin sensitivity (H-IS) is crucial. In this study, we investigated the factors that may preserve H-IS in patients with NAFLD in longitudinal observations. Comprehensively, putative biomarkers in blood and urine, body composition, tissue-specific lipid accumulation and insulin sensitivity by 1H-MRS and hyperinsulinemic euglycemic clamp, respectively, and pathological diagnosis using liver biopsy were assessed in patients with NAFLD (n=62) at baseline. After more than one year of general treatment, consented 36 patients (average age 51.3±11.2, men n=21, diabetes n=23) were subjected to the same assessment as baseline. To investigate the indices associated with H-IS, all indices were compared between upper and lower one third patients of Δ% suppression of endogenous glucose production (EGP) (+11.5±4.0 vs. -10.9±5.1%, respectively). As results, body weight, fasting plasma glucose, HbA1c, muscle and adipose tissue insulin sensitivity, visceral fat, and adiponectin levels were significantly improved in improved H-IS group. Interestingly, Δhepatic lipid content or fibrosis index was not different between the improved and worsened H-IS groups. Furthermore, simple regression analysis with Δ% suppression of EGP and all indices revealed that Δ% suppression of free fatty acid, visceral fat area, and adiponectin levels were significantly associated with Δ% suppression of EGP. Multiple regression analysis with Δ% suppression of EGP and these three indices showed that only Δ% suppression of free fatty acid significantly associated with Δ% suppression of EGP. Taken together, not only lipid volume but also the quality of accumulated lipid is important to preserve hepatic insulin sensitivity in patients with NAFLD.

Disclosure

F.Shigiyama: None. S.Hiruma: None. N.Kumashiro: Research Support; Terumo Corporation, LifeScan Diabetes Institute, Sumitomo Dainippon Pharma Co., Ltd., Speaker's Bureau; Daiichi Sankyo, Novo Nordisk.
158-OR: Ceramides Predict the Development of Type 2 Diabetes
Authors: LEIHERER ANDREAS , MUENDLEIN AXEL , PLATTNER THOMAS , VONBANK ALEXANDER , MADER ARTHUR , SPRENGER LUKAS , MAECHLER MAXIMILIAN , LARCHER BARBARA , FRAUNBERGER PETER , DREXEL HEINZ , SAELY CHRISTOPH H.
Published: June 2023

Ceramides recently have attracted interest as a marker of cardiovascular event risk. However, only little is known about their role in the development of type 2 diabetes (T2DM). We analyzed the ceramide ratio Cer d18:18/Cer d 18:24:0 (CerRatio) in 894 consecutive Caucasian patients who were referred to angiography for the evaluation of established or suspected stable coronary artery disease (CAD). T2DM was diagnosed according to ADA criteria. The incidence of T2DM was recorded over a follow-up time of up to 16 years (median: 13 [8-14] years). At baseline, the CerRatio was significantly higher in T2DM (n=239) than in nondiabetic patients (4.6e-2 vs. 3.9e-2, p<0.001). During follow-up, diabetes was newly diagnosed in 71 patients, i.e. in 11% of the initially nondiabetic subjects. The CerRatio strongly predicted the incidence of T2DM both univariately (standardized OR 1.47 [1.11-1.96]; p=0.008) and in a multivariately adjusted model (figure) comprising age, sex, BMI, the presence of significant CAD at angiography, and hemoglobin A1c (HbA1c; OR=1.45 [1.07-1.98]; p=0.018). Though the CerRatio was significantly correlated with BMI (r=0.138, p<0.001) and HbA1c (r=0.137, p<0.001), applying a mediation analysis revealed that BMI and HbA1c had only marginal indirect effects (5, 7% and 3.6%, respectively) on the association of the CerRatio with T2DM incidence. We conclude that ceramides strongly and independently predict the development of diabetes.

Disclosure

A. Leiherer: None. A. Muendlein: None. T. Plattner: None. A. Vonbank: None. A. Mader: None. L. Sprenger: None. M. Maechler: None. B. Larcher: None. P. Fraunberger: None. H. Drexel: None. C.H. Saely: None.
1580-P: Role of Hepatic Glycogen on Nocturnal Gluconeogenesis in Type 2 Diabetes
Authors: UNNI UMA , YADAV YOGESH , MUGLER JOHN P. , CARTER RICKEY , BASU ANANDA , BASU RITA
Published: June 2023

Higher gluconeogenesis (GNG) during the latter part of the night contributes to higher endogenous glucose production (EGP) in type 2 diabetes (T2D). We have shown that glycogen synthesis is reduced in T2D due to a functional defect in the rate-limiting hepatic glucokinase activity. Studies using MRIs have confirmed lower net hepatic glycogen content in T2D than subjects without diabetes (ND) before and after dinner. We hypothesize that increasing post dinner hepatic glycogen content might restore overnight EGP in T2D by reducing GNG. 14 T2D and 15 anthropometrically matched ND were randomly assigned to glycogen loading (GL; 60% carb) vs. non glycogen loading (NGL; 40% carb) meals for 3 days. [6,6-2H2] Glucose was infused to measure EGP. GNG was estimated with deuterium labeled water and glycogen content measured via C13 NMR in fed and fasted state. Results are shown in Figure1. Liver glycogen content was higher with GL vs NGL in both cohorts. EGP was higher (p<0.05) in GL vs NGL throughout night in both cohorts. The percent contribution of GNG and glycogenolysis (GGL) to EGP averaged ~ 50% in ND throughout the night in both GL and NGL. In contrast, the percent contribution of GNG to EGP in T2D with GL was lower vs. NGL overnight (average ~48% vs. 60%) and matched the pattern observed in ND with GL. The results indicate that lower glycogen content in the liver contributes to increased nocturnal rates of GNG in T2D.

Disclosure

U.Unni: None. Y.Yadav: None. J.P.Mugler, iii: None. R.Carter: None. A.Basu: None. R.Basu: Consultant; Sparrow Pharmaceuticals Inc, Research Support; Abbott Diabetes, AstraZeneca.

Funding

National Institute for Health and Care Research (R01DK029953)
1583-P: Dynamic Tracking and Identification of Liver-Specific Secretory Proteins in the Circulation of Live Mice
Authors: KIM KWANG-EUN , MYOUNG SUH JAE
Published: June 2023

Secretory proteins are an essential component of interorgan communication networks that regulate animal physiology. Current approaches for identifying secretory proteins from specific cell and tissue types are largely limited to in vitro or ex vivo models which often fail to recapitulate in vivo biology. As such, there is mounting interest in developing in vivo analytical tools that can provide accurate information on the origin, identity, and spatiotemporal dynamics of secretory proteins. Here, we describe iSLET (in situ Secretory protein Labeling via ER-anchored TurboID) which selectively labels proteins that transit through the classical secretory pathway via catalytic actions of Sec61b-TurboID, a proximity labeling enzyme anchored in the ER lumen. To validate iSLET in a whole-body system, we express iSLET in the mouse liver and demonstrate efficient labeling of liver secretory proteins which could be tracked and identified within circulating blood plasma. Furthermore, proteomic analysis of the labeled liver secretome enriched from liver iSLET mouse plasma is highly consistent with previous reports of liver secretory protein profiles. Taken together, iSLET is a versatile and powerful tool for studying spatiotemporal dynamics of secretory proteins, a valuable class of biomarkers and therapeutic targets.

Disclosure

K.Kim: None. J.Suh: None.

Funding

National Research Foundation of Korea (2018R1A2A3075389, 2016M3A9B6902871, 2017K1A1A2013124, 2021R1A2C2007573, 2020R1C1C1013927); KAIST Key Research Institutes Project; Bio and Medical Technology Development Program (2019M3E5D3073104); Ministry of Science
1588-P: Translatable Modeling of Glucagon Actions in Human iPSC-Derived Hepatocytes Uniquely Predicts Differential Clinical Responses to Glucagon Analog Agonist Molecules
Authors: ROELL WILLIAM , REGMI AJIT , BENSON CHARLES T. , COSKUN TAMER , MOYERS JULIE S. , ALSINA-FERNANDEZ JORGE , THOMAS MELISSA K.
Published: June 2023

As the development of clinically efficacious glucagon receptor (GCGR) agonists for the treatment of metabolic diseases evolves, the challenge to identify physiologically relevant preclinical models that predict glucagon receptor actions in humans becomes ever more important. While many hepatocyte models are available, most fail to recapitulate fundamental metabolic regulatory responses to key physiologic stimuli including integration of glucagon and insulin actions that govern hepatic carbohydrate or lipid metabolism. We developed and optimized a human induced pluripotent stem cell (iPSC)-derived hepatocyte model to mimic physiologic insulin and glucagon signaling and counter-regulation of metabolic pathways. We leveraged the model to evaluate functional responses of different glucagon receptor agonist molecules to regulate carbohydrate and lipid metabolism. Unlike assays conducted in several other cell line and primary isolated hepatocyte systems, hepatic glucose output assays in the human IPSC hepatocyte model demonstrated substantial potency differences between two novel soluble glucagon analog agonists. These findings predicted differential dose-dependent blood glucose excursions in response to these agonists when administered in a single ascending dose study in healthy human volunteers. Modeling physiologic integration of insulin and glucagon signaling in a clinically translatable human iPSC hepatocyte model can advance understanding of the regulation of nutrient metabolism by glucagon receptor agonists in humans and support the development of novel therapeutic approaches to address metabolic disorders, including type 2 diabetes, obesity, and nonalcoholic fatty liver disease.

Disclosure

W.Roell: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. A.Regmi: Employee; Eli Lilly and Company. C.T.Benson: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. T.Coskun: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. J.S.Moyers: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. J.Alsina-fernandez: Employee; Eli Lilly and Company. M.K.Thomas: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company.
1589-P: Portal Serotonin Is Critical for Efficient Hepatic Control of Systemic Glycemia
Authors: KIEDROWSKI MATTHEW , EL SANADI CAROLINE , PENN MARC S. , GEHO WALTER
Published: June 2023

There is growing evidence for a portal factor, released following a meal, that combined with insulin, induces hepatic glycogen synthesis. We have developed an in vitro model that demonstrates that serotonin could be the portal factor. Using rat primary hepatocyte cultures in 6 well plates with 750,000 cells per well and a 4-hour treatment time, we observed that 0.1 mU/ml of human insulin resulted in generation of 8.8 ± 2.3 ng glycogen/ug total protein compared to 6.7 ± 1.0 ng glycogen/ug total protein, with 5 uM 5HT-serotonin. Interestingly, the combination of 0.1 mU insulin/ml and 5 uM 5HT-serotonin resulted in 22.5 ± 3.8 ng glycogen/ug total protein, a 256% increase in glycogen synthesis compared to insulin alone. In the absence of serotonin, it takes approximately 10-fold greater insulin to achieve the same levels of glycogen synthesis as insulin with serotonin. These data combined with the observation that 95% of serotonin is made in the gut, that gastrointestinal serotonin is released upon food ingestion and the growing evidence that obese subjects have decreasing levels of circulating serotonin, suggest that gastrointestinal serotonin could be the portal factor necessary for efficient hepatic control of system glycemia. These data further suggest that insulin insensitivity is due to loss of a critical co-factor binding, and that this co-factor is serotonin. In summary, type 1 diabetes is due to a deficiency of insulin and type 2 diabetes may be due to a deficiency of its co-factor, portal serotonin.

Disclosure

M.Kiedrowski: None. C.El sanadi: None. M.S.Penn: Consultant; Diasome, Quest Diagnostics. W.Geho: None.
159-LB: Circulating MicroRNAs as Predictors of ß-Cell Function in Youth Onset Type 2 Diabetes—The TODAY Study
Authors: TRYGGESTAD JEANIE B. , BIALEK SHANNAN , REDLING DAKOTA , CHERNAUSEK STEVEN , EL GHORMLI LAURE
Published: June 2023

In the Treatment Options for Type 2 Diabetes in Adolescents and Youth (TODAY) study, half the participants reached the primary outcome (HbA1c≥8% for at least 6 months) within 4 years which was associated with a decrease in C-peptide oral disposition index (oDI). We aimed to identify circulating miRNA species associated with β cell function decline. Following a preliminary survey of select participants using nCounter Human v3 miRNA Panel (NanoString Technologies), polymerase chain reaction validations were carried out for 17 miRNAs from 365 participants with samples at baseline, 24, 60, 96, and 120 months. During the first 6 months in TODAY, 150 (43%) of the participants (mean age 14 years at baseline and diabetes duration <2 years) had a ≥20% decrease in C-peptide oDI. Using backward elimination modeling and adjusting for demographic characteristics, three baseline miRNA log2 fold changes remained significant predictors of this C-peptide oDI decline ≥20% (p<0.05). Increased levels of miR-155 (OR:1.21, 95%CI:1.07-1.38) and miR-130b (OR:1.32, 95%CI: 1.02-1.70) were associated with oDI decline, while decreased levels of miRNA-126 (OR:0.60, 95%CI: 0.44-0.81) were associated with oDI decline. miR-122 was negatively correlated with C-peptide oDI at baseline and 24-months (R=0.22, p<0.01 and R=0.19, p<0.01, respectively), and positively correlated with proinsulin, at baseline, 24-, and 60-months (R=0.26, p<0.01, R=0.26, p<0.01, R=0.18, p<0.01, respectively). These miRNA species are associated with alterations in β cell insulin secretion and apoptosis targeting pyruvate dehydrogenase, glucokinase, and GLI-similar zinc finger proteins, suggesting that differences in baseline abundance may serve as circulating markers of β cell dysfunction, providing potential mechanistic insights into the aggressive nature of youth-onset type 2 diabetes.

Disclosure

J. B. Tryggestad: None. S. Bialek: None. D. Redling: None. S. Chernausek: Advisory Panel; Ascendis Pharma A/S. L. El ghormli: None.

Funding

National Institutes of Health (5U01DK061230-16)
159-OR: Greater Sleep Variability Is Associated with Less Time in Range of Glucose Levels in Patients with Type 1 Diabetes
Authors: MARTYN-NEMETH PAMELA , ABU IRSHEED GHADA , PARK MINSUN , STEFFEN ALANA D. , BURKE LARISA , PRATUANGTHAM SARIDA , BARON KELLY , DUFFECY JENNIFER , PEREZ ROSANNE , QUINN LAURIE T. , CLARK WITHINGTON MEG , SALEH ADAM H. , LOIACONO BERNARDO , MIHAILESCU DAN , REUTRAKUL SIRIMON
Published: June 2023

Background: Sleep and circadian disturbances emerge as novel factors influencing glycemic control in type 1 diabetes (T1D). We aimed to explore the associations between sleep, behavioral circadian parameters, self-care, and glycemic parameters in T1D.

Methods: Baseline data of 76 non-shift working adult T1D patients participating in a randomized study to optimize sleep health (NCT04506151) were analyzed. Glycemic assessments were derived from blinded 7-day continuous glucose monitoring (CGM) and A1C. Percentages of time-in-range (TIR of glucose levels between 70-180 mg/dL) and %CV were calculated from CGM. Sleep was recorded using 7-day actigraphy, yielding sleep duration and efficiency. Variability (SD) of mid-sleep time (MST) was used to represent sleep variability. Non-parametric behavioral circadian variables were derived from actigraphy activity recordings. Self-care was measured by the Diabetes Self-Management Questionnaire (DSMQ-R). Multiple regression analyses were performed to identify independent predictors of glycemic parameters.

Results: Mean (SD) age was 36.3 (11) years, 48 (63.2%) female, and median (IQR) A1C 6.8% (6.2, 7.4). Sleep duration, efficiency, and non-parametric behavioral circadian variables were not associated with glycemic parameters. After adjusting for age, sex, insulin delivery mode/CGM use, and ethnicity, each hour increase in SD of MST was associated with 9.64 % less TIR (B= -9.64, 95% (CI) [-16.29, -2.99], p=<0.001). Higher DSMQ score was an independent predictor of lower A1C (B = -0.18, 95% CI -0.32, -0.04).

Conclusion: Greater sleep variability is independently associated with less time spent in desirable glucose range in this modern T1D cohort, with clinically important effect sizes. Sleep optimization to reduce variable sleep schedule could potentially lead to improved metabolic control and should be explored in future research.

Disclosure

P.Martyn-nemeth: None. L.T.Quinn: None. M.Clark withington: None. A.H.Saleh: None. B.Loiacono: None. D.Mihailescu: None. S.Reutrakul: Speaker's Bureau; Eli Lilly and Company. G.Abu irsheed: None. M.Park: None. A.D.Steffen: None. L.Burke: None. S.Pratuangtham: None. K.Baron: Other Relationship; National Sleep Foundation, Research Support; Google. J.Duffecy: Consultant; Chorus Health, K Health, Stock/Shareholder; Healent Health, My Alloy. R.Perez: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (R01DK121726)
1592-P: Evaluating Metabolic Effects of Nicotinamide Riboside on Hepatic Substrate Metabolism in CDAA-Induced NAFLD Mice
Authors: SHEN QINGYANG , PUGMIRE JACKSON K. , SUWANNASUAL USA , MAPTUE NESMINE R. , KHEMTONG CHALERMCHAI
Published: June 2023

Nonalcoholic fatty liver disease (NAFLD) is associated with insulin resistance and altered hepatic energy metabolism, with imbalanced glucose and lipid metabolism. Nicotinamide riboside (NR) has recently emerged as a potential therapeutic for a broad range of metabolic abnormalities. Here, we evaluate the effect of NR treatment on hepatic substrate metabolism in mice with NAFLD. NAFLD was induced by feeding the animals with a choline-deficient, amino acid-define (CDAA) diet for 4 weeks. Mice (C57BL/6) in the NR treatment group (CDAA+NR) were then put on a NR-containing CDAA diet for 10 weeks while the non-treated animals remained on CDAA. Both groups of animals had similar food consumption and weight gain rates, with comparable liver-to-body weight ratios. At the conclusion of the treatment, isolated livers were perfused via the portal vein with 13C metabolic tracers, i.e. 5.5 mM [1,6-13C]glucose + 0.4 mM [U-13C]long chain fatty acids (LCFA) suspended in bovine serum albumin. Fractional substrate oxidations and flux analyses were done using 13C nuclear magnetic resonance (NMR) spectroscopy. An increasing trend of oxygen consumption rates was observed as a result of NR treatment (1.50±0.43 µmol/min/liver) compared to the CDAA-treated group (0.68±0.15 µmol/min/liver). However, the increase is not statistically different. 13C NMR isotopomer flux analyses revealed that pyruvate dehydrogenase flux relative to citrate synthase flux are comparable across the 2 groups (CDAA: 30±4%; CDAA+NR: 33±10%). No changes in fractional oxidation of 13C-long-chain fatty acids and 13C-glucose were observed as a result of nicotinamide riboside treatment, with 44±2% and 48±3% of Ac-CoA derived from 13C-LCFA in CDAA and CDAA+NR livers, respectively. 13C-glucose contributed as a minor source of Ac-CoA (~4-5%) in both groups. These findings suggest that nicotinamide riboside treatment did not affect substrate, fats vs glucose, utilization in CDAA-induced NAFLD mouse livers.

Disclosure

Q. Shen: None. J.K. Pugmire: None. U. Suwannasual: None. N.R. Maptue: None. C. Khemtong: None.

Funding

University of Florida
1594-P: Interorgan Insulin-Leptin Signal Cross Talk Originating in the Liver Enhances Survival under Food-Shortage Conditions
Authors: TAKAHASHI KEI , YAMADA TETSUYA , KATAGIRI HIDEKI
Published: June 2023

The liver can store energy and distribute it to other tissues/organs, according to the energy states involving hepatic insulin action. In addition, we and others have shown that the liver transmits metabolic information to other organs, via neuronal and humoral mechanisms. Herein, we demonstrate a novel form of inter-organ crosstalk originating in the liver, which links hepatic insulin action with hypothalamic leptin signaling and promotes survival during food shortage. We first observed that tamoxifen-inducible liver insulin receptor (IR) knockout (iLIRKO) mice ate more and consumed less oxygen with decreased hypothalamic leptin action, thereby gaining body weight. Circulating soluble leptin receptor (sLepR) levels were elevated in iLIRKO mice. We then produced tamoxifen-inducible liver leptin receptor (LepR) and IR double-knockout (iLLepRIRKO) mice. In iLLepRIRKO mice, sLepR elevations were not seen. Moreover, increased body weights, hyperphagia and decreased whole-body oxygen consumption seen in iLIRKO mice were reversed, indicating the importance of hepatic sLepR production in the liver-hypothalamus crosstalk. Human hepatic lepr and circulating sLepR levels also correlated negatively with hepatic insulin action indices. Additionally, we restricted food intake in wild type mice. Food restriction decreased hepatic insulin action and whole-body oxygen consumption with increased circulating sLepR. We furthermore performed food restriction in tamoxifen-inducible liver LepR knockout (iLLepRKO) mice. Notably, in iLLepRKO mice, the decreases in basal metabolic rates were blunted and mortality was markedly increased during food restriction. Collectively, the liver translates metabolic cues regarding energy-deficient status, which is reflected by decreased hepatic insulin action, into sLepR, thereby suppressing energy dissipation. This novel inter-organ crosstalk may assure survival of individuals facing food shortages in nature.

Disclosure

K.Takahashi: Speaker's Bureau; Taisho Pharmaceutical Holdings Co., Ltd. T.Yamada: Speaker's Bureau; MSD, Mitsubishi Tanabe Pharma Corporation, Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd., Abbott Japan Co., Ltd. H.Katagiri: None.

Funding

Japan Society for the Promotion of Science (21K08522, 22H03126, 20H05694); Japan Science and Technology Agency (JPMJPS2023)
1595-P: Empagliflozin Improves Nephropathy and Heart Failure with Preserved Ejection Fraction Despite Limited Effects on NASH in a Novel Obese Diabetic Hamster Model
Authors: BRIAND FRANCOIS , DUBROCA CAROLINE , SULPICE THIERRY
Published: June 2023

Introduction: Sodium-glucose cotransporter 2 inhibitor empagliflozin (EMPA) has shown significant benefits for patients with diabetic nephropathy and heart failure with preserved ejection fraction (HFpEF), but the effects on nonalcoholic steatohepatitis (NASH) and liver fibrosis remain unclear. To further investigate the benefits of EMPA on these comorbidities, we established a novel diet-induced obese diabetic hamster model with human-like NASH and lipoprotein cholesterol metabolism, that also develops nephropathy and HFpEF.

Methods: Obesity, NASH, and HFpEF were induced with a 20-week free choice diet, which presents hamsters with a choice between control chow or high fat/cholesterol diet, and normal water or 10% fructose water. After 20 weeks of diet, obese hamsters were intraperitoneally injected with streptozotocin (STZ) to induce diabetes and were then treated orally once daily for 5 weeks with vehicle or EMPA 30mg/kg.

Results: Compared with vehicle, EMPA reduced hyperglycemia (vehicle: 373 +/- 43 mg/dL; EMPA: 182 +/- 30 mg/dL, p<0.01), increased pancreatic insulin content (+146%, p<0.05), significantly reduced plasma triglycerides and moderately raised LDL-cholesterol levels, as expected. However, EMPA did not improve NASH (including hepatocyte ballooning score) and liver fibrosis. EMPA reduced glomerular filtration rate and proteinuria (p=0.06 for both vs. vehicle), and significantly reduced urine albumin-to-creatinine ratio (-35%, p<0.01). In addition, EMPA showed significant improvement in HFpEF with reduced E/A and E’/E ratios with no alteration in ejection fraction, as measured by echocardiography.

Conclusion: EMPA improves kidney parameters and HFpEF despite limited effects on NASH in obese diabetic hamsters. This preclinical model with multiple comorbidities will be useful to evaluate the combination of EMPA or other antidiabetic therapies with drugs targeting NASH.

Disclosure

F. Briand: Employee; PHYSIOGENEX. Stock/Shareholder; PHYSIOGENEX. C. Dubroca: Employee; CARDIOMEDEX. T. Sulpice: Employee; PHYSIOGENEX. Stock/Shareholder; PHYSIOGENEX. Employee; CARDIOMEDEX. Stock/Shareholder; CARDIOMEDEX.
1597-P: KCNH6 Regulates Hepatic Mitochondrial Function and Glucose Metabolism via NF-κB Pathway
Authors: ZHAO RUXUAN , LU JING , XIONG FENGRAN
Published: June 2023

KCN, a human ether-a-go-go-related gene, encodes KCN ion channel in the Kv11 family, which ranks 11th in abundance among different ion channel families. KCNH6, a subfamily of KCN potassium channel family, has been proven to affect glucose metabolism and insulin secretion both in humans and mice. However, the precise function of KCNH6 in the liver remains unknown. In this study, we showed that knockout (KO) of Kcnh6 impairs mitochondrial function and insulin tolerance as well as promotes high-fat-diet-induced insulin resistance. In line, overexpression of Kcnh6 in hepatocytes ameliorates fatty acid oxidation, mitochondrial respiration, and the abundance of mitochondrial respiratory complexes ameliorating insulin resistance. Similarly, induction of Kcnh6 by tail vein injection into KO mice decreased insulin tolerance. Mechanistically, Kcnh6 improved mitochondrial function and insulin secretion via the the NFκB-IκB kinase (IKK) pathway both in vitro and in vivo. Therefore, we suggest Kcnh6 as a potential novel therapeutic target to treat hepatic energy metabolism and mitochondrial dysfunction and insulin resistance.

Disclosure

R.Zhao: None. J.Lu: None. F.Xiong: None.
1598-P: MIF-upregulated CD74 in Liver Contributes to the Development of NAFLD
Authors: CHEN LIUJUN , LI LISHA , HUANG YIHENG , CHEN XI , QI YADAN , TONG HAIBIN , WU HONG , QI DAKE
Published: June 2023

Nonalcoholic fatty liver disease (NAFLD) is associated with metabolic dysfunctions, such as obesity. Macrophage migration inhibitory factor (MIF), a pro-cytokine, was also identified to regulate NAFLD, but the molecular mechanisms remain unclear. We presently indicated that MIF has a non-inflammatory effect on triggering NAFLD through upregulating its cell membrane receptor, CD74. MIF increased CD74 protein rather than gene expression in liver cells in a time-dependent manner. That was not associated with any changes in the expression of pro-inflammatory factors, such as TNF-α, IL-6 and IL-1β. In a MIF overexpressed animal model (Mif lung Tg), high circulating MIF levels increased hepatic CD74 proteins but not genes in the absence of inflammation in the liver. High fat diet also increased circulating MIF levels leading to an upregulation of hepatic CD74. All these animal models with high circulating MIF and hepatic CD74 were accompanied with steatosis and fibrosis in the liver. These alterations could be reversed by either MIF neutralization or MIF knockout. Caspase 4 facilitates the degradation of CD74. MIF endocytosis inhibited caspase 4 cleavage and activation. Thus, inhibition of MIF upregulated cleaved caspase 4 leading to a degradation in CD74. Overall, our findings indicate that MIF stabilizes CD74 through suppressing caspase 4 and this may be an important mechanism in regulating NAFLD in the absence of inflammation.

Disclosure

L. Chen: None. L. Li: None. Y. Huang: None. X. Chen: None. Y. Qi: None. H. Tong: None. H. Wu: None. D. Qi: None.

Funding

Canadian Institutes of Health Research (PJT156116); Research and Development Corporation; National Sciences and Engineering Research Council of Canada (RGPIN-2017-04542); National Institutes of Health (AR-078334); China Scholarship Council (to L.C., Y.H., L.L.)
16-LB: Teneligliptin Observational Retrospective Real-World Study to Evaluate Its Effect on Renal Parameters and Safety in Indian Type 2 Diabetes Mellitus Patients with Chronic Kidney Disease (TOP RENAL)
Authors: SURYAWANSHI SACHIN , JADHAO VIBHUTI , BRID MANASI , BHUSHAN SUMIT , BARKATE HANMANT V.
Published: June 2023

Background and Aim: Teneliglitpin, approved in India since 2015, is the most commonly prescribed DPP4i in India. Due to lack of evidence, this study was planned in real world setting to evaluate effects of teneligliptin on renal parameters and safety in Indian T2DM patients with CKD.

Methodology: TOP RENAL real world study enrolled T2DM patients with eGFR ≤60 to ≥15 ml/min/1.73m2 and were started on teneligliptin (20/40 mg OD) and continued it for next six months. Those patients prescribed any antidiabetic medication along with teneligliptin or those with change in antidiabetic prescription in the following six months were excluded.

Results: 377 patients data was collected during August'22-October'22 and grouped according to baseline eGFR (ml/min/1.73m2): G3a: 45-59 (n=102); G3b: 30-44 (n=124); G4: 15-29 (n=151). The mean age was 59.86±9.58 years and 47% were females. Renal parameters: eGFR, Sr creatinine and BUN were assessed at baseline, 3 months and 6 months. Results showed non-significant changes in eGFR (Table1), Sr. creatinine and BUN at 3 and 6 months in all three groups. No safety concerns or hypoglycemia were reported during the study.

Conclusion: Teneligliptin initiation in Indian T2DM patients with mild-severe renal impairment did not show any safety concerns and was well tolerated.

Disclosure

S. Suryawanshi: Employee; Glenmark Pharmaceuticals. V. Jadhao: Employee; Glenmark Pharmaceuticals. M. Brid: Employee; Glenmark Pharmaceuticals. S. Bhushan: None. H. V. Barkate: None.

Funding

Glenmark Pharmaceuticals Limited
160-LB: Circulating MicroRNAs as Predictors of Microvascular Complications in Youth Onset Type 2 Diabetes—The TODAY Study
Authors: REDLING DAKOTA , BIALEK SHANNAN , CHERNAUSEK STEVEN , EL GHORMLI LAURE , TRYGGESTAD JEANIE B.
Published: June 2023

From the Treatment Options for Type 2 Diabetes in Adolescents and Youth (TODAY) study, 60% of the participants demonstrated diabetes-related complications by 10 years. We aimed to identify circulating microRNA (miRNA) species associated with microvascular disease in TODAY participants with youth-onset type 2 diabetes. Using nCounter Human v3 miRNA Panel (NanoString Technologies) to identify relevant miRNAs in two groups of participants who lost glycemic control and either developed microvascular complications during follow-up (n=6) or not (n=11), 53 miRNA species were differentially expressed at baseline (p<0.05) and 2 at the end of study visit (p<0.05). Polymerase chain reaction validations were carried out for 17 miRNAs from 365 participants with samples obtained at baseline, 24, 60, 96, and 120 months. During follow-up, 250 (68%) of the participants (mean 14 years old and diabetes duration <2 years at entry) developed at least one microvascular complication. Using backward elimination modeling and adjusting for demographic characteristics, a set of four baseline miRNA log2 fold changes remained significant predictors of microvascular complications (p<0.05). Increased levels of miR-122 (OR:1.17, 95%CI:1.03-1.32) and miR-221 (OR:1.51, 95%CI: 1.51-1.98) were associated with higher risk of complication, while increased levels of miRNA-let 7g (OR:0.75, 95%CI: 0.60-0.94) and miRNA-130a (OR:0.82, 95%CI: 0.68-0.99) were associated with lower risk of complication. miR-122 abundance was also positively correlated with urine albumin/creatinine ratio at baseline and 24-months (R=0.11, p=0.041 and R=0.16, p=0.005, respectively). The alterations in miR-122, 221, 130a, and let 7g in circulation are all consistent with in vitro alterations that promote endothelial dysfunction suggesting that these early alterations in circulation may predict those at highest risk of microvascular disease in the future.

Disclosure

D. Redling: None. S. Bialek: None. S. Chernausek: Advisory Panel; Ascendis Pharma A/S. L. El ghormli: None. J. B. Tryggestad: None.

Funding

National Institutes of Health (5U01DK061230-16)
1608-P: Phenotype of Liver-Specific MC3R Knockout Mice
Authors: CHEN JOCELYN , PATEL TUSHAR P. , JAIN ARAD , ELIZONDO DIANA , ALTMAN ELIZABETH K. , TUGARINOV NICOL , HOLLIS MELANIE , ROBERSON ROBIN , CARON ALEXANDRE , ELMQUIST JOEL K. , YANOVSKI JACK A.
Published: June 2023

The melanocortin 3 receptor (MC3R) is a key regulator of energy homeostasis and feeding behavior. Global Mc3r knockout mice exhibit greater fat mass and reduced lean mass. Our previous research has found that Mc3r transcripts can be detected in liver and the MC3R deficiency phenotype is partially reversed in liver-specific MC3R recovery mice on a chow diet, in both male and female mice, and high fat diet, in females only. To further examine the role of hepatic MC3R, we produced a liver-specific Mc3r knockout model by crossing mice expressing albumin-Cre recombinase (Alb-Cre+/+) to mice with a floxed Mc3r gene (LoxP+/+), and placed them on a 45 kcal% energy from fat diet for 12 weeks. Female liver-specific Mc3r-deficient mice with diet-induced obesity displayed nonsignificant trends towards worsened glucose tolerance during a glucose tolerance test (glucose AUC Lox+/+, Alb-Cre+/+: n=10, 24036±1644; Alb-Cre+/+: n=10, 19745±1364; BL/6: n=11, 21072±1152 min*mg/dL, p=0.059) and insulin resistance during a glucose tolerance test (glucose AUC LoxP+/+, Alb-Cre+/+: n=10, 14938±812.2;Alb-Cre+/+: n=14, 13062±850; BL/6: n=11, 11725±1024 min*mg/dL, p=0.12), but no differences in morning serum C-peptide (LoxP+/+, Alb-Cre+/+: n=8, 0.74±0.07; Alb-Cre+/+: n=4, 0.55±0.15; BL/6: n=8, 0.29±0.05 ng/mL, p=0.32). Similar patterns were observed for total food consumption (LoxP+/+, Alb-Cre+/+: n=10, 37.2±1.2; Alb-Cre+/+: n=15, 34.5±0.7; BL/6: n=10, 35.7±1.4 g/15 days, p=0.082), with no differences in feeding efficiency (LoxP+/+, Alb-Cre+/+: n=10, 0.027±0.008; Alb-Cre+/+: n=15, 0.009±0.011; BL/6: n=10, 0.017±0.025 g/kcal, p=0.19). No notable trends were observed in male mice. These data suggest that hepatic Mc3r deficiency alone is at most a minor contributor to the obesity phenotypes associated with HFD-treated Mc3r-deficient mice. Additional analysis of tissue samples from these mice and future studies examining Mc3r function in the liver will further elucidate the role of hepatic MC3R.

Disclosure

J.Chen: None. J.K.Elmquist: Research Support; Regeneron Pharmaceuticals Inc. J.A.Yanovski: Research Support; Hikma Pharmaceuticals, Rhythm Pharmaceuticals, Inc., Soleno Therapeutics, Inc., Versanis Bio Inc. T.P.Patel: None. A.Jain: None. D.Elizondo: None. E.K.Altman: None. N.Tugarinov: None. M.Hollis: None. R.Roberson: None. A.Caron: None.
1609-P: Neutral Sphingomyelinase Inhibition Limits Hepatic-Lipotoxicity and Inflammation in a Cell Model of Steatosis
Authors: ALRASHED FATEMA , AREFANIAN HOSSEIN , SINDHU SARDAR T. , BAHMAN FATEMAH , ALSAEED HALEMAH , AL MADHOUN ASHRAF , ALZAID FAWAZ , AL-MULLA FAHD , AHMAD RASHEED
Published: June 2023

Nonalcoholic fatty liver disease (NAFLD) represents a global healthcare challenge; it is the hepatic manifestation of metabolic syndrome and is strongly associated with developing type 2diabetes mellitus (T2DM). Liver fat accumulation is the first step of disease progression that triggers hepatic lipotoxicity. The role of ceramides in inducing deleterious effects on hepatic metabolism is now well-accepted. Yet, the specific role of stress responsive sphingomyelinase activation under lipotoxic conditions remains under-investigated. The complexity of NAFLD pathogenesis contributes to lack of proper investigatory model, limiting progression in developing and testing novel treatment and prevention strategies. Here, we first report a convenient in-vitro human cell-based model with great resemblance to in-vivo NAFLD hallmarks. Neutral Sphingomyelinase (nSMase2) expression and activity was found to be elevated in both the liver of high fat steatosis mouse models and in HepG2-steatosis cell models. Meanwhile, the functional inhibition of nSMase2 prevented hepatotoxicity-induced pathologies by significantly reducing intracellular lipid accumulation and prevented the upregulation of TNF-α triggered inflammation. Furthermore, inhibition of nSMase2 showed significant increase in PPARα at both gene and protein levels, while PPARα reduction was observed under the stimulation of nSMase2 activity by its agonist daunorubicin (DNR). Together the presented data highlight the role of nSMase2 in the pathogenesis of NAFLD and other disorders linked to hepatic steatosis, providing a novel therapeutic target.

Disclosure

F. Alrashed: None. H. Arefanian: None. S.T. Sindhu: None. F. Bahman: None. H. AlSaeed: None. A. Al Madhoun: None. F. Alzaid: None. F. Al-Mulla: None. R. Ahmad: None.

Funding

Kuwait Foundation for the Advancement of Sciences (RA0402021)
1610-P: Fatty Acid–Binding Protein 4 as a Biomarker for Remission of Type 2 Diabetes
Authors: STENTZ FRANKIE B.
Published: June 2023

Elevated levels of circulating Fatty Acid Binding Protein 4 (FABP4), also named adipocyte FABP (A-FABP), are associated with obesity, diabetes mellitus, insulin resistance, hypertension, and cardiovascular risk. FABP4 is a protein found in high levels in both macrophages and adipocytes. Release of FABP4 from adipocytes via lipolysis, and release from macrophages during inflammation, plays a key role in development of insulin resistance and diabetes mellitus. We studied the effect of a high protein (HP; 30% protein, 40% carbohydrate, 30% fat) diet vs high carbohydrate (HC;15% protein, 55% carbohydrate, 30% fat) diet in patients recently diagnosed with Type 2 Diabetes (T2D). We determined the levels of circulating FABP4 and metabolic parameters in the HP vs HC groups at baseline and 6 months after consuming the respective diets. Participants (14) were randomized to a HP or HC diet for 6 months. All food was provided and designed for weight loss. After 6 months on the HP diet, 100% of the subjects had remission of T2D to Normal Glucose Tolerance (NGT), whereas only 16.6% of subjects on HC diet had remission of T2D. The HP diet group exhibited significant improvement in a) insulin sensitivity (p=0.001), b) inflammatory cytokines (p=0.001), C) cardiovascular risk factors (p=0.004) compared to the HC diet group at 6 months. Although weight loss in both groups was around 10% at 6 months, the HP group lost % fat mass, whereas, the HC group lost both % muscle and fat mass. Average level of FABP4 decrease after 6 months was 12,831 pg/ml in the 8 participants on the HP diet, compared to only a 1183 pg/ml average decrease after 6 months for the 6 HC participants. Conclusions: These data showed a significant decrease of circulating FABP4 in the HP but not HC diet group and suggests that a high protein diet could be an optimal way to reduce FABP4 levels in the blood. This could subsequently lower the risk of development/progression of insulin resistance, T2D, inflammation and cardiovascular risk conditions and serve as a biomarker.

Disclosure

F.B.Stentz: None.

Funding

AD Baskin
1615-P: The Peripheral and Central Cross Talk Mediated by Adipose MIF—A Novel Mechanism of Hyperphagia following Olanzapine Treatment
Authors: CHEN XI , HUANG YIHENG , QI YADAN , CHEN LIUJUN , LI LISHA , GAO PINGYI , CUI DONGHONG , QI DAKE
Published: June 2023

Olanzapine is one of the first-line antipsychotic medications but it usually induces metabolic side effects, characterized by obesity. We previously indicated that olanzapine leads to hyperphagia through the effects of a proinflammatory cytokine, macrophage migration inhibitory factor (MIF) in the hypothalamus. However, whether olanzapine's upregulated peripheral MIF could affect the central mechanism is currently unknown. Our present study found that olanzapine increases ectopic expression of dopamine receptor 1 (DRD1) in adipocytes which relatively upregulates MIF expression and release through a PKA/CREB signaling pathway. In global MIF knockout mice, MIF accumulation appears in the hypothalamus following peripheral injection of recombinant MIF proteins, suggesting that peripheral MIF indeed travels to the hypothalamus. Mif Lung Tg mice (MIF overexpression in the lung) with increased circulating MIF levels had hyperphagia and activation of the CaMKKβ/AMPK/AgRP signaling pathway in the hypothalamus. Overall, our study for the first time proved that adipose-derived MIF is a crucial factor in regulating metabolic side effects induced by olanzapine.

Disclosure

X.Chen: None. Y.Huang: None. Y.Qi: None. L.Chen: None. L.Li: None. P.Gao: None. D.Cui: None. D.Qi: None.

Funding

National Sciences and Engineering Research Council of Canada (RGPIN-2017-04542); Canadian Institutes of Health Research (PJT-156116); China Scholarship Council (202106230212)
1622-P: Complex Mechanisms Underlie the Antidiabetic Effect of Tempol in Experimentally Induced Diabetes Mellitus
Authors: NAGUIB YAHYA
Published: June 2023

Diabetes mellitus (DM) is a vastly growing worldwide health problem which imposes public health, social, as well as economic burden. Multifactorial and complicated pathophysiological pathways may underlie DM. Attention has been given for the search of polymodal therapeutic molecules. Herein, we tested the capacity of tempol, a known superoxide dismutase-mimetic in the treatment of experimentally induced DM. Thirty male Wistar rats weighing 150-200 g were equally divided into: 1- Control group (rats received a single intra-peritoneal injection of citrate buffer for 4 weeks), 2- Untreated diabetic group, and 3- Tempol treated diabetic group. At the end of the study blood samples were collected for the measurement of fasting blood glucose, fasting blood insulin, liver function, kidney function, oxidative stress markers, and inflammatory markers. Pancreas, skeletal muscles, and liver were removed for histopathological, immunohistochemical and RT-PCR studies. Treatment with tempol significantly improved glycemic status, insulin secretion, sensitivity and reserve. Tempol preserved both liver and kidney functions. Tempol also significantly enhanced energy status, and countered structural changes, inflammation, and apoptosis of pancreatic tissue. Tempol significantly upregulated pancreatic AMPK, and skeletal muscle and hepatic glucose transporters. Interestingly, tempol demonstrated an endogenous stem cells recruitment capacity. In conclusion, tempol exhibited a potential capacity to counter the complexity of the underlying mechanisms in DM.

Disclosure

Y.Naguib: None.
1623-P: Identification of Gut Microbes That Prevents Metabolic Syndrome in Healthy Japanese Adults
Authors: HONOKI HISAE , FUJISAKA SHIHO , WATANABE YOSHIYUKI , OKU MAKITO , KONDO YUKI , NISHIMURA AYUMI , KADO TOMONOBU , BILAL MUHAMMAD , ENKAKU ASAKO , TAKIKAWA AKIKO , CHUJO DAISUKE , MORINAGA YOSHITOMO , TOBE KAZUYUKI
Published: June 2023

Gut microbiota is an important driver of metabolic status. Recent studies have demonstrated that dysbiosis precedes the development of metabolic disorders. However, the bacterial species associated with metabolic syndrome and their physiological role in obesity and glucose metabolism have not been fully elucidated. In this study, we performed 16S rRNA sequencing analysis on fecal samples from 49 metabolically healthy volunteers, a detailed lifestyle questionnaire survey including diet, and an analysis on metabolic parameters. We also examined the metabolic effects of some microbes by transplantation into HFD-fed mice. The average age, BMI and HbA1c were 43.5±9.0, 23.9±3.9 and 5.5±0.3, respectively. A clear correlation was observed between lifestyle and metabolic parameters. Analysis of fecal microbial structure divided the participants into three enterotypes according to their predominant phylogenic pattern; 1) Prevotella copri group, 2) Bacteroides group, and 3) others. Enterotype 1 had higher random blood glucose (BG), and lower HDL-C, while enterotype 2 had lower random BG, BMI, and waist circumference. Eleven bacterial species were identified as being positively or negatively correlated with metabolic parameters. Relative abundances of the Family Fusobacteriaceae and Prevotella stercorea were positively correlated with BMI, fasting BG, and visceral fat mass. In contrast, the Family Rikenellaceae and Bacteroides uniformis (Bu) negatively correlated with metabolic parameters. Transplantation of Alistipes indistinctus (Ali), a major bacterium of the Family Rikenellaceae, or Bu into HFD-fed C57BL/6J mice suppressed body weight gain, improved glucose tolerance and histological findings without altering food intake. In summary, our results suggest that microbial community structure can be altered by lifestyle even in good health. Moreover, maintaining microbial species such as Ali and Bu may contribute to the prevention of metabolic disorders in Japanese adults.

Disclosure

H. Honoki: None. S. Fujisaka: None. Y. Watanabe: None. M. Oku: None. Y. Kondo: None. A. Nishimura: None. T. Kado: None. M. Bilal: None. A. Enkaku: None. A. Takikawa: None. D. Chujo: None. Y. Morinaga: None. K. Tobe: Other Relationship; MSD K.K., Novo Nordisk Pharma Ltd., Takeda Pharmaceutical Company Limited, Daiichi Sankyo, Mitsubishi Tanabe Pharma Corporation, Suntory Global Corporation, Ltd.,, Taiho Pharmaceutical Co. Ltd., Japan Diabetes Foundation, Japan Association for Diabetes Education and Care.

Funding

Japan Society for the Promotion of Science (20K10318); Japan Association for Diabetes Education and Care