bluuebunny/crossref_metadata_2025 huggingface dataset:
358-OR: Adiponectin, Leptin, and FGF-21 Levels in Adolescents Exposed and Not Exposed to Gestational Diabetes—Results from the Danish National Birth Cohort
Authors: KAMPMANN FREJA BACH , BAUN THUESEN ANNE CATHRINE , HJORT LINE , BJERRE METTE , FRYSTYK JAN , TETENS INGE , BJERREGAARD ANNE AHRENDT , CHAVARRO JORGE E. , OLSEN SJURDUR F. , DAMM PETER , VAAG ALLAN A. , GRUNNET LOUISE
Published: June 2019

Background: Offspring of mothers with gestational diabetes mellitus (GDM) are at increased risk of developing metabolic diseases. Adipokines such as adiponectin and leptin may be involved in the underlying mechanisms. Fibroblast growth factor 21 (FGF-21) has recently been identified as a potent metabolic regulator. Therefore, we investigated the impact of fetal exposure to GDM on adiponectin, leptin and FGF-21 levels and their associations with metabolic and adiposity traits during adolescence.

Methods: A prospective study comparing 502 offspring of GDM exposed mothers and 535 offspring of non-exposed mothers. During a clinical exam of offspring, aged 9-16 years old, metabolic traits were measured and fasting blood samples collected and assayed for serum concentrations of leptin, adiponectin and FGF-21.

Results: GDM exposed offspring had 34% (95% CI: 18-51%) higher leptin levels, and 34% (95% CI: -48, -15%) lower FGF-21 concentrations than non-exposed offspring (p<0.05), but no significant differences in adiponectin concentrations were found. These differences remained significant after adjusting for major confounders, including maternal pre-pregnancy BMI. Offspring leptin concentrations correlated positively with insulin and HOMA-IR, whereas adiponectin concentrations were inversely correlated (p<0.05). Elevated leptin and decreased adiponectin concentrations correlated with all measures of adiposity, whereas FGF-21 correlated with total fat percentage only. In analyses stratified by offspring BMI z-score categories, GDM exposure had no significant effect on leptin and adiponectin concentrations.

Conclusion: GDM exposed offspring had higher leptin and lower FGF-21 concentrations than non-exposed controls, but this was driven by higher prevalence of adiposity among GDM offspring. The differential adipokine concentration does not seem to be the major driver behind the dysmetabolic phenotype among GDM offspring.

Disclosure

F.B. Kampmann: None. A. Thuesen: None. L. Hjort: None. M. Bjerre: None. J. Frystyk: None. I. Tetens: None. A.A. Bjerregaard: None. J.E. Chavarro: None. S.F. Olsen: None. P. Damm: Advisory Panel; Self; Novo Nordisk A/S. Other Relationship; Self; Novo Nordisk A/S. A.A. Vaag: Employee; Self; AstraZeneca. L. Grunnet: None.

Funding

Danish Council for Strategic Research; Innovation Fund, Denmark (09-067124, 11-115923); Novo Nordisk Foundation (NNF160C0022518); Danish Diabetes Academy
362-OR: Deletion of METTL14 in Beta Cells in Adult Mice Induces Diabetes Related to Increased ER Stress
Authors: MEN LILI , SUN JUAN , REN DECHENG
Published: June 2019

N6-methyladenosine (m6A) is the most abundant mRNA modification. However, the role of m6A in regulating β-cell function is unknown. To address the functional role of METTL14, a key component of the m6A methyltransferase complex, in β-cells, we generated an METTL14 β-cell specific knockout mouse in adult MIP-CreERT transgenic mice. 4 weeks after tamoxifen treatment (100 mg/kg for 3 times every other day), the feeding glucose of βKO mice were 55.1% higher in βKO mice. βKO mice are also glucose intolerant. The AUC was increased by 54.7% in βKO mice (P<0.01). The fasting glucose were 307.5 ± 24.3 mg/dl in βKO mice and 219.0 ± 7.8 mg/dl in control mice (P<0.001). The response to insulin was similar between βKO and control mice. The feeding insulin levels in βKO mice were significantly decreased by 58.9% (P<0.01). The serum insulin levels from βKO mice were significantly lower at 0, 15 and 30 minutes after glucose challenge. In addition, the insulin secretion from βKO islets was less than 50% of control islets after incubated at 16.7 mM glucose. Surprisingly, β-cell mass in βKO mice was 55.9% higher than that in control mice (P<0.05). In addition, we did not detect β-cell death in βKO islets. However, β-cell proliferation in βKO islets was increased by 1.3-fold. To understand the molecular mechanism, we performed RNA-seq to detect the gene expression in βKO and control islets from male mice 8 weeks after tamoxifen treatment. Among 106 altered genes in βKO islets, 104 genes were upregulated (P<0.001) and 2 genes were downregulated. The genes response to ER stress such as Pdia4, Itpr1, Thbs1, Ire1α and Igfbp5 were among the top up-regulated genes. We applied m6A-seq in MIN6 cells and confirmed that Ire1α was modified by m6A in its last exon and 3' UTR region. We further showed that both mRNA and protein of Ire1α and spliced XBP-1 were significantly increased in βKO islets.

In conclusion, METTL14 plays an important role in regulating insulin secretion, and METTL14 deficiency in β-cells induces diabetes by increasing ER stress.

Disclosure

L. Men: None. J. Sun: None. D. Ren: None.

Funding

National Institutes of Health (P30DK020595)
365-OR: The NLRP3 Inflammasome Mediates Early Beta-Cell Dysfunction Triggered by Islet Amyloid
Authors: DENROCHE HEATHER C. , KIM NAHAE , YAU KIANA , NACKIEWICZ DOMINIKA , ORBAN PAUL C. , SUEN IMELDA , VERCHERE CAMERON B.
Published: June 2019

Islet amyloid, formed by islet amyloid polypeptide (IAPP) aggregates, may be a trigger of islet inflammation in T2D. In vitro studies using bone marrow-derived macrophages suggest roles for Toll-like receptor 2 (TLR2), and the NLRP3 inflammasome in mediating IAPP aggregate-induced inflammatory gene expression. To assess the impact of amyloid on islet macrophages in vivo, we performed single cell RNA sequencing on sorted islet immune cells from mice which develop islet amyloid via expression of human IAPP (hIAPP-Tg mice); this uncovered robust changes in islet macrophage populations, including upregulation of TLR2 (∼3.0 fold, p=0.002) and NLRP3 (∼2.8 fold, p=0.006) in proinflammatory macrophages. To test whether these receptors mediate IAPP aggregate-induced inflammation and beta cell dysfunction in vivo, we examined if TLR2 or NLRP3 deletion attenuated islet amyloid-induced beta cell dysfunction by crossing hIAPP-Tg mice to Tlr2-/- or Nlrp3-/- mice. Loss of TLR2 did not impact beta cell dysfunction in hIAPP-Tg Tlr2-/- male or female mice compared to hIAPP-Tg Tlr2+/+ littermates, revealing that TLR2 is not necessary for islet-amyloid driven beta cell pathology. In contrast, NLRP3 knockout improved glucose tolerance in young hIAPP-Tg male mice and delayed diabetes onset on high fat diet (HFD) (10.9 vs. 4.9 weeks; p=0.02) compared to hIAPP-Tg littermates with intact NLRP3. However, hIAPP-Tg Nlrp3-/- males eventually developed diabetes and glucose intolerance on long-term HFD. In female hIAPP-Tg mice, in which islet amyloid forms more slowly, NLRP3 deletion markedly improved glucose tolerance after 18 weeks of HFD relative to hIAPP-Tg Nlrp3+/+ controls (AUC: 886 ± 207 vs. 1854 ± 296, p<0.0001). The glycemic benefit of NLRP3 knockout was not due to altered insulin sensitivity, and was absent in non-hIAPP expressing mice, which do not form amyloid. Thus, NLRP3, but not TLR2, mediates early islet amyloid-induced beta cell dysfunction, and may be a therapeutic target for islet inflammation in T2D.

Disclosure

H.C. Denroche: Advisory Panel; Self; Integrated Nanotherapeutics. N. Kim: Employee; Self; Integrated Nanotherapeutics. K. Yau: None. D. Nackiewicz: None. P.C. Orban: None. I. Suen: None. C.B. Verchere: Advisory Panel; Self; Sirona Biochem. Board Member; Self; Integrated Nanotherapeutics.

Funding

JDRF; Canadian Institutes of Health Research
371-P: Hypoglycemia in the Brain Is Required to Augment Counterregulatory Responses to Hypoglycemia when Liver Glycogen Levels Are Enhanced
Authors: WARNER SHANA O. , SMITH MARTA S. , FARMER BEN , WINNICK JASON J.
Published: June 2019

In a previous study, we showed that elevated hepatic glycogen levels can generate a signal in the liver during insulin (INS)-induced hypoglycemia that is relayed to the brain, resulting in enhanced counterregulatory (CR) hormone secretion and net hepatic glucose output (NHGO). The purpose of this study was to determine whether hypoglycemia in the brain is required for this augmentation to occur. Fasted (18 hour) dogs were studied and each experiment began with a 4h glycogen deposition period in which each animal received somatostatin and basal intraportal infusions of both INS and glucagon (GGN). Plasma glucose (PG) was doubled by an intravenous (IV) glucose infusion and liver glycogen content was enhanced in all animals (76 ± 6 and 68 ± 4 mg/g in Brain EU and HYPO, respectively) by an intraportal infusion of fructose. A 2h control period followed, during which hyperglycemia and basal replacement of hormones were continued but fructose infusion was stopped. In the final 2h hypoglycemic period, somatostatin and GGN infusions were stopped and the plasma INS level was increased 16-fold by an IV infusion of INS. During the final period, whole body PG was clamped at 52 ± 1 mg/dL in one group (HYPO; n=6). In a second group, PG in the head was clamped at 92 ± 3 mg/dL by infusing glucose into the carotid and vertebral arteries (Brain EU; n=8) while the glucose level in the rest of the body remained at 51 ± 1 mg/dL. During the final hour of the hypoglycemic period, NHGO was lower in Brain EU compared to HYPO (0.8 ± 0.2 vs. 2.2 ± 0.4 mg/kg/minute, p<0.01). This decrease in NHGO was associated with a decrease in CR hormone secretion, as GGN (27 ± 6 vs. 53 ± 8 pg/mL, p<0.01) and epinephrine (451 ± 103 vs. 1106 ± 143 pg/mL, p<0.01) were lower when the brain was euglycemic (Brain EU vs. HYPO, respectively). These data suggest that the augmented CR responses that occur during INS-induced hypoglycemia when liver glycogen content is enhanced, require hypoglycemia in the brain.

Disclosure

S.O. Warner: None. M.S. Smith: None. B. Farmer: None. J.J. Winnick: None.

Funding

National Institutes of Health
38-OR: Beta-Arrestin-1 Regulates Pancreatic Beta-Cell Function and Plays a Key Role in Maintaining Whole Body Glucose Homeostasis
Authors: BARELLA LUIZ F. , ROSSI MARIO , PYDI SAI PRASAD , JAIN SHANU , ZHU LU , CHEN WEI , WESS JÜRGEN
Published: June 2019

Beta-arrestin-1 and -2 (barr1 and barr2, respectively) are major GPCR-associated signaling molecules that are widely expressed throughout the body. Studies with whole body barr1 and barr2 KO mice have shown that beta-arrestins play important roles in key metabolic functions. In this study, we inactivated the barr1 gene in a conditional fashion specifically in beta-cells of adult mice (beta-barr1-KO mice). In vivo studies showed that beta-barr1-KO mice exhibited a pronounced impairment in glucose tolerance and glucose-stimulated insulin secretion (GSIS), when mice were maintained on high-fat diet (HFD). Studies with isolated perifused pancreatic islets prepared from HFD beta-barr1-KO mice confirmed that barr1 deletion in beta-cells led to a significant decrease in GSIS. Interestingly, HFD beta-barr1-KO mice displayed pronounced reductions in beta-cell mass and the rate of beta-cell proliferation, suggesting that barr1 is required for the expansion of beta-cell mass that occurs during HFD feeding. To test whether enhanced barr1 signaling in beta-cells might ameliorate the metabolic deficits associated with the consumption of a HFD, we also generated transgenic mice over-expressing barr1 in beta-cells (RIPII-barr1 mice). These transgenic mice displayed metabolic phenotypes that were opposite to those observed with the beta-barr1-KO mice, including improved glucose tolerance and GSIS in vivo as well as increased beta-cell mass. This is the first study demonstrating that barr1 is critical for the proper function of pancreatic beta-cells and for maintaining whole body glucose homeostasis. Identification of the cellular mechanisms through which barr1 signaling promotes beta-cell function and proliferation may lead to the development of novel drugs useful for the treatment of type 2 diabetes.

Disclosure

L.F. Barella: None. M. Rossi: None. S. Pydi: None. S. Jain: None. L. Zhu: None. W. Chen: None. J. Wess: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases
380-P: Online CME Improves Clinical Knowledge/Competence Related to Managing T2D in Older Patients with Hypoglycemic Risk
Authors: LARKIN AMY , LACOUTURE MICHAEL , LE ANNE
Published: June 2019

We sought to determine if online continuing medical education (CME) activities could improve the clinical knowledge/competence of primary care physicians (PCPs), diabetologists/endocrinologists (D/Es), and nurses/nurse practitioners (N/NPs) regarding the management type 2 diabetes (T2D) in older patients with hypoglycemia risk. The effect of two online, video-based CME activities were evaluated using knowledge/competence linked pre-/post-assessment questions. McNemar’s chi-squared test (P <.05 indicates statistical significance) assessed educational effect. The activities launched between August 23, 2018 and November 1, 2018, and data were collected for 8 weeks each. In total, 202 D/Es, 1,192 PCPs and 5,480 N/NPs answered all pre-/post-assessment questions for at least 1 activity. Improvements were demonstrated among D/Es, PCPs, and N/NPs: Knowledge: 35% of D/Es (N=139, P<.0001), 40% of PCPs (N=729, P<.0001), and 42% of N/NPs (n=4119, P< .0001) improved regarding insulin dosing when switching between insulin therapies Competence: 27% of D/Es (N=63, P=.0023), 33% of PCPs (N=463, P<.0001), and 18% of N/NPs (n=1361, P< .0001) improved related to selection of antihyperglycemic agent for an older T2D patient at risk of hypoglycemia 63% of D/Es (N=139, P<.0001), 56% of PCPs (N=729, P<.0001), and 43% of N/NPs (n=4119, P< .0001) improved related to the most appropriate insulin dose for a given patient Continued educational gaps: • 32% of D/Es (N=63), 44% of PCPs (N=463), and 63% of N/NPs (n=1361) failed to select the most appropriate insulin for a patient at risk of hypoglycemia • 33% of D/Es (N=63), 37% of PCPs (N=463), and 43% of N/NPs (n=1361) failed to recognize the impact on hypoglycemia of switching basal insulin therapies This study demonstrates the success of online video-based CME on improving knowledge/competence in D/Es, PCP, and N/NPs related to managing T2D in older patients. Continued knowledge gaps were identified for future educational targets.

Disclosure

A. Larkin: None. M. LaCouture: None. A. Le: None.

Funding

Sanofi
387-P: N-acetyl Cysteine Does Not Prevent Impaired Awareness of Hypoglycemia
Authors: MOHEET AMIR , KUMAR ANJALI , COLES LISA , RUBIN NATHAN , EBERLY LYNN E. , SEAQUIST ELIZABETH R.
Published: June 2019

Impaired awareness of hypoglycemia (IAH) is common in diabetes. Administration of N-acetyl cysteine (NAC) during episodes of hypoglycemia (HG) prevents the development of IAH in rodents (Fioramonti 2013). Here we tested the hypothesis that NAC will be effective in preventing IAH in healthy humans using a double blind cross over study design where subjects received either NAC or saline (S) in random order on day 1 of a 2 day protocol that involved 3 HG clamps. On day 1, 350 mg NAC or S was given IV over 5 hours starting 30 min before start of IV insulin (2 mg/kg/min). Blood glucose (BG) was targeted at 50 mg/dl for 2 hours, then at 95 mg/dl for 2 hours, then 50 mg/dl for 2 hours. After BG recovery subjects were sent home to return on day 2 for a final 2-hour HG clamp. During the 1st and 3rd HG periods, serum epinephrine (EPI) and hypoglycemic symptoms (HSx) were collected. Subjects received 25 mg diphenhydramine IV at 0 and 4 hours due to a high occurrence of drug reactions. Additional doses or odansetron were given as needed.

EPI during clamp 1 was higher during NAC compared to S but the HSx were the same. No difference was seen between subjects who did or did not have a reaction to NAC. EPI and HSx on day 2 following NAC were lower than on day 1 but no different from response measured following S on day 1. NAC elicited many adverse events, enhanced EPI during HG, and did not alter EPI or HSx during HG on day 2 compared to S. Further development of IV NAC as a therapy for IAH is unwarranted.

Disclosure

A. Moheet: None. A. Kumar: None. L. Coles: None. N. Rubin: None. L.E. Eberly: None. E.R. Seaquist: Advisory Panel; Self; Eli Lilly and Company, Zucara Therapeutics Inc. Consultant; Self; 360 Consulting, MannKind Corporation. Research Support; Self; Eli Lilly and Company, JDRF, National Institutes of Health. Other Relationship; Self; American Diabetes Association, Sanofi, WebMD.

Funding

JDRF
4-LB: Feeding-Stimulated Regulation of the Epigenome Controls Adaptive Insulin Secretion
Authors: WORTHAM MATTHEW , LIU FENFEN , FLEISCHMAN JOHANNA , WALLACE MARTINA , HARRINGTON AUSTIN R. , CHIOU JOSHUA , GAULTON KYLE , METALLO CHRISTIAN , SANDER MAIKE
Published: June 2019

Adaptation of the insulin secretory response to fluctuating insulin demand is critical for blood glucose homeostasis. We postulated that the epigenome could link changes in metabolic state with adaptive insulin secretion. Here we show that perturbations of nutrient state lead to extensive changes to H3K27 acetylation in pancreatic islets. Genomic sites of H3K27ac deposition acutely regulated by feeding were enriched for binding by the histone demethylase Lsd1, a candidate nutrient sensor known to respond to hormonal and metabolic stimuli. Conditional deletion of Lsd1 in mouse β-cells resulted in hypoglycemia and fasting hyperinsulinemia. Lsd1-deficient β-cells exhibited changes in H3K4me1 and H3K27ac deposition at Lsd1-bound sites. These changes at the chromatin level associated with aberrant expression of nutrient signaling and metabolic genes. We further show that Lsd1 controls feeding-stimulated transcription downstream of cAMP, which has been implicated in the control of β-cell metabolism. Accordingly, Lsd1-deficient β-cells failed to attenuate glucose metabolism in the fasted state, resulting in basal insulin hypersecretion. Further supporting a role for Lsd1 in adaptive insulin secretion, islets from hyperinsulinemic db/db mice exhibited increased H3K27ac and H3K4me1 levels at Lsd1-bound sites. Moreover, genomic sites bound by LSD1 in human islets were enriched for genetic variants associated with fasting glucose and type 2 diabetes in genome-wide association studies, suggesting a conserved role of LSD1 in the regulation of insulin secretion. Combined, our findings provide evidence that nutrient state modulates adaptive insulin secretion through Lsd1-dependent changes to the epigenome.

Disclosure

M. Wortham: None. F. Liu: None. J. Fleischman: None. M. Wallace: None. A.R. Harrington: None. J. Chiou: None. K. Gaulton: None. C. Metallo: Employee; Spouse/Partner; ICON plc. Research Support; Self; Gilead Sciences, Inc., Nimbus Therapeutics. M. Sander: None.

Funding

National Institutes of Health (R01DK068471); JDRF (3-PDF-2014-193-A-N)
402-P: Higher Glucose Thresholds for Hypoglycemia Alarms on Continuous Glucose Monitoring Systems Correlate with Less Time in Hypoglycemia
Authors: LIN YU KUEI , HUNG MAN , SHARMA ANU , CHAN OWEN , VARNER MICHAEL , GROAT DANIELLE , DOPITA DANA , FISHER SIMON
Published: June 2019

Based on the set glucose thresholds for hypoglycemia alarms, continuous glucose monitoring (CGM) systems can provide audible alerts that notify individuals for actual/impending hypoglycemic events. While CGMs have been shown to reduce hypoglycemic events in patients with type 1 diabetes (T1D), there are little data demonstrating the association between glucose thresholds for hypoglycemia alarms and clinical hypoglycemia outcomes. This cross-sectional observational study enrolled 83 T1D patients with CGM use >3 months, active CGM usage time ≥70% and activated (i.e., turned “on”) hypoglycemia alarms. CGM settings and two-week CGM glucose data (total 25,683 hours) were analyzed. Glucose thresholds for hypoglycemia alarms (mean: 74 ± 8 mg/dL; range: 60-100 mg/dL) correlated inversely with time in hypoglycemia (both glucose <54 and <70 mg/dL) (Figure 1a). When comparing the cohort with hypoglycemia alarm threshold settings at glucose level ≤70 mg/dL (n=42), threshold settings ≥75 mg/dL (n=41) were associated with a 74% and 60% reduction in hypoglycemia time (<54 and <70 mg/dL, respectively) (Figure 1b). All subjects with hypoglycemia alarm set at glucose ≥75 mg/dL spent less than 3% of time with glucose <54 mg/dL. The current data support the use of higher glucose thresholds on CGM hypoglycemia alarms to decrease the time in hypoglycemia in patients with T1D.

Disclosure

Y. Lin: None. M. Hung: None. A. Sharma: None. O. Chan: Research Support; Self; Zucara Therapeutics Inc. M. Varner: None. D. Groat: None. D. Dopita: None. S. Fisher: None.
406-P: Effect of Metformin on Dicarbonyl Stress in Patients with Type 2 Diabetes and Chronic Heart Failure
Authors: KOPECKY JAN , HOŠKOVÁ EVA , VELEBA JIRI , MALINSKA HANA , VELEBOVA KATERINA , MELENOVSKY VOJTECH , PELIKANOVA TEREZIE
Published: June 2019

Background and Aims: Treatment with metformin (MET) in diabetic patients with chronic heart failure (HF) is associated with better cardiovascular outcomes. The endothelial dysfunction, atherosclerosis and macrovascular diabetic complications are associated with excessive generation of reactive dicarbonyls like methylglyoxal (MG). MET has been proposed as a scavenger of methylglyoxal. The aim of the study was to evaluate the effects of MET on substrate metabolism and dicarbonyl stress in patients with DM and HF.

Materials and Methods: A randomized, double-blind, placebo-controlled, crossover study in a total time of six months testing the effect of 3- month usage of MET (2 g/day) vs. placebo. A group of 40 treatment naive patients with type 2 DM and HF were included. At the beginning and at the end of each intervention period various metabolic tests were done including hyperinsulinemic-isoglycemic clamp, parameters of dicarbonyl stress like MG, Glutathione (GSH), glutathione disulfide (GSSG), and the activity of Glyoxalase1 (GLO-1).

Results: MET significantly reduced HbA1c compared to placebo (p˂0.01). Insulin sensitivity, measured as a metabolic clearance rate of glucose (MCR) during the clamp, improved significantly with MET (p˂0.001) but also, although less significantly, with placebo (p˂0.05), difference between MET and placebo was not significant. Compared to placebo, GHS levels increased after the intervention (baseline vs. MET vs. PL: 866 ± 231 vs. 994 ± 342 vs. 878 ± 266µmol/l, p<0.05). GSH/GSSG ratio was not affected. The levels of MG decreased after MET (p<0.05), decrease after placebo was not significant. The activity of GLO-1 was not affected.

Conclusions: Our results on a unique group of patients with heart failure and diabetes support the evidence that one of the cardioprotective pathways of metformin is through reducing dicarbonyl stress. As there was no change in the GLO-1 activity, it might be due to binding and deactivation of reactive dicarbonyls by MET.

Disclosure

J. Kopecky: None. E. Hošková: None. J. Veleba: None. H. Malinska: None. K. Velebova: None. V. Melenovsky: None. T. Pelikanova: None.

Funding

Agentura pro zdravotnický výzkum Ceské Republiky (AZV16-27496A)
409-P: Cardiac Biomarkers and Incident Peripheral Arterial Disease (PAD) in Patients with Type 2 Diabetes in the ADVANCE Trial
Authors: HICKS CAITLIN W. , WANG DAN , MATSUSHITA KUNIHIRO , CHALMERS JOHN , WOODWARD MARK , SELVIN ELIZABETH
Published: June 2019

Background: High-sensitivity cardiac troponin T (hs-cTnT) and natriuretic peptide (NT-proBNP) are strongly associated with coronary heart disease, heart failure, and death in population-based studies. Little is known about the associations of these cardiac biomarkers with incident PAD in adults with type 2 diabetes.

Methods: We used Cox models to assess the associations of elevated baseline hs-cTnT (≥ 14 ng/L) and NT-proBNP (≥ 200 pg/mL) in participants in the ADVANCE trial with measured biomarkers and without PAD at baseline. We also modeled hs-cTnT and NTproBNP using restricted cubic splines.

Results: Incident PAD occurred in 417 of 6,428 ADVANCE participants during a median follow-up of 5 years. After multivariable adjustment, increasing baseline hs-cTnT and NT-proBNP were both associated with PAD (Figure). Modeled categorically, PAD risk was higher in participants with elevated hs-cTnT (HR 1.35, 95% CI 1.07-1.70) or NT-proBNP (HR 1.44, 95% CI 1.15-1.81). Participants with elevations of both biomarkers had the highest risk (HR 1.85, 95% CI 1.38-2.49).

Conclusion: Hs-cTnT and NT-proBNP were associated with incident PAD in adults with diabetes even after adjusting for cardiovascular risk factors. Cardiac biomarkers may be useful to identify persons with type 2 diabetes at risk for future PAD.

Disclosure

C.W. Hicks: None. D. Wang: None. K. Matsushita: Consultant; Self; Akebia, Kyowa Hakko Kirin Co., Ltd. Research Support; Self; Fukuda Denshi, Kyowa Hakko Kirin Co., Ltd. J. Chalmers: Research Support; Self; Idorsia. M. Woodward: Consultant; Self; Amgen Inc., Kirin. E. Selvin: None.
414-P: Insulin Secretion Capacity Relates to Human Ventricular Myocardial Mitochondrial Function
Authors: ZWECK ELRIC , SCHEIBER DANIEL , ALBERMANN SOPHIE , BORGER JULIUS , JELENIK TOMAS , PESTA DOMINIK , HORN PATRICK , BOEKEN UDO , AKHYARI PAYAM , KELM MALTE , RODEN MICHAEL , WESTENFELD RALF , SZENDROEDI JULIA
Published: June 2019

Hyperinsulinemia and impaired insulin secretion are potential contributors to the development of diabetes-related heart failure. Interference with ventricular myocardial mitochondria might play a key pathogenic role, but human data confirming these connections are limited. We hypothesized that ventricular myocardial mitochondrial function is impaired in hyperinsulinemia but is positively related to insulin secretion capacity. We recruited 32 heart transplant recipients without heart failure or antihyperglycemic treatment (age: 53.5±13.3years; BMI: 24.8±2.9kg/m²; HbA1c: 5.9±0.7%). We measured serum insulin levels and blood glucose every 30 minutes for three hours after an oral 75g glucose uptake. Seven patients were newly diagnosed with type 2 diabetes and six had impaired glucose tolerance. High-resolution respirometry was performed on transcatheter ventricular endomyocardial biopsies to determine mitochondrial oxidative capacity, coupling efficiency (respiratory control ratio, RCR) and intrinsic uncoupling (leak control ratio, LCR). Fasting serum insulin, a marker of insulin resistance, was inversely associated with ventricular myocardial mitochondrial RCR (r=-0.39; p=0.02) and positively with LCR (r=0.42; p=0.02), but not with oxidative capacity (p=0.34). Maximum increase of insulin in proportion to fasting levels strongly related to ventricular myocardial mitochondrial RCR (r=0.62; p=0.0001) and inversely to LCR (r=-0.46; p=0.01). This insulin secretory response related to fatty acid-dependent respiration (r=0.35; p=0.05) and tended to relate to combined oxidative capacity from fatty acid and glycolytic substrates (r=0.31; p=0.09). Maximum insulin response following glucose ingestion relates to cardiac mitochondrial efficiency and oxidative capacity, while insulin resistance might affect cardiac energy metabolism. These data suggest a pathogenic role of hyperinsulinemia as well as beta cell failure in diabetes-related heart failure.

Disclosure

E. Zweck: None. D. Scheiber: None. S. Albermann: None. J. Borger: None. T. Jelenik: None. D. Pesta: None. P. Horn: None. U. Boeken: None. P. AKhyari: Research Support; Self; Abbott, Edwards Lifesciences Corporation, Medtronic. M. Kelm: None. M. Roden: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Poxel SA, Servier. Board Member; Self; Eli Lilly and Company. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi. Speaker's Bureau; Self; Novo Nordisk Inc. R. Westenfeld: None. J. Szendroedi: None.

Funding

German Research Foundation (CRC1116); Heinrich-Heine University
419-P: LDL Particle Size Is a Predictor of Cardiovascular Events in Cardiovascular Disease Patients with Nonalcoholic Fatty Liver Disease as Well as in Those without Nonalcoholic Fatty Liver Disease
Authors: LEIHERER ANDREAS , MUENDLEIN AXEL , SAELY CHRISTOPH H. , VONBANK ALEXANDER , LARCHER BARBARA , MADER ARTHUR , DOPHEIDE JÖRN F. , BAUMGARTNER IRIS , FRAUNBERGER PETER , DREXEL HEINZ
Published: June 2019

Nonalcoholic fatty liver disease (NAFLD) is associated with atherogenic dyslipidemia, in particular with a small LDL particle size. However, there are no data on the association between LDL particle size and cardiovascular events in cardiovascular disease patients with NAFLD. This issue is addressed in the present study. We measured LDL particle size and concentration in 298 statin-naïve cardiovascular disease patients, of whom 141 had NAFLD. In patients with NAFLD, LDL particle size was significantly lower (21.0 vs. 21.2 nm, p<0.001) and particle concentration was significantly higher (1487 vs. 1336 nmol/l, p=0.006) than in those without NAFLD. Prospectively, we recorded 135 cardiovascular events during a follow-up time of 8.2±3.3 years. In multivariate Cox regression analysis LDL particle size (standardized adjusted HR 0.68 [0.54-0.85], p=0.001), but not LDL particle concentration (standardized adjusted HR 0.97 [0.79-1.19], p=0.774) was a significant and independent predictor of cardiovascular events after adjustment for age, sex, BMI, hypertension, smoking, HDL particle size and HDL particle concentration, and for the presence of T2DM and NAFLD. In subgroup analysis regarding the presence of NAFLD, LDL particle size significantly predicted cardiovascular events both in patients with NAFLD (standardized adjusted HR 0.60 [0.42-0.86], p=0.005) and in those without NAFLD (standardized adjusted HR 0.64 [0.46-0.90], p=0.010). We conclude that LDL particle size is a predictor of cardiovascular events in cardiovascular disease patients with NAFLD as well as in those without NAFLD.

Disclosure

A. Leiherer: None. A. Muendlein: None. C.H. Saely: None. A. Vonbank: None. B. Larcher: None. A. Mader: None. J.F. Dopheide: None. I. Baumgartner: None. P. Fraunberger: None. H. Drexel: None.
42-LB: A New Laminopathy Caused by an Arg133/leu Mutation in Lamin A/C and the Effects thereof on Adipocyte Differentiation and the Transcriptome
Authors: WANG ZHE , HE YINGZI , ZHENG FENPING
Published: June 2019

Mutations in LMNA trigger changes in nuclear architecture, signal transduction, and gene regulation, associated with a group of multisystem diseases termed laminopathies. Here we report a new laminopathy in a 26-year-old female, characterized by generalized lipoatrophy, insulin-resistant diabetes, micrognathia, symptoms of premature aging, hypertension, and biopsy-proven focal segmental glomerulosclerosis. The de novo heterozygous mutation R133L lamin A/C gene (LMNA) was found in the patient and inherited to her son who has not (yet) developed a lipodystrophic phenotype. We analyzed the nuclear morphology and laminar distribution of both 3T3-L1 pre-adipocytes and mouse glomerular mesangial cells overexpressing human wild type lamin A/C (LMNA WT) or lamin A/C with the R133L mutation (LMNA R133L) that causes the new laminopathy. We found that nuclear morphology was abnormal; the nuclei evidenced hypertrophy, nuclear membrane invagination or blebbing, and a disorganized A-type lamin meshwork in both cell lines overexpressing LMNA R133L. The differentiation from 3T3-L1 pre-adipocyte into adipocytes was impaired in cells expressing LMNA R133L; the expression levels of genes associated with adipose tissue self-renewal, including adipogenesis and angiogenesis were downregulated. Furthermore, the insulin-signaling pathway was inhibited in LMNA R133L adipocytes. Microarray gene expression profiling showed that the most prominent differences between 3T3-L1 cells expressing LMNA WT and LMNA R133L were in genes implicated in metabolic pathways, the cellular response to DNA damage, DNA repair, and RNA binding. We thus expanded the clinical spectrum of laminopathy and concluded that the LMNA R133L mutation associated with lipodystrophic features, multisystem disorders and the repairation of DNA damage.

Disclosure

Z. Wang: None. Y. He: None. F. Zheng: None.

Funding

Zhejiang Provincial Natural Science Foundation of China (Y17H070002); National Natural Science Foundation of China (81471099)
42-OR: Hub Cells Orchestrate 3-Dimensional Pancreatic Beta-Cell Ca2+ Dynamics In Vivo
Authors: SALEM VICTORIA , DELGADILLO SILVA LUIS F. , SUBA KINGA , ALONSO ALDARA MARTIN , KIM WHEI-CHANG , KALOGIANNI VASILIKI , AKHTAR NADEEM , GHARAVY NEDA MOUSAVY , GEORGIADOU ELENI , LECLERC ISABELLE , BRIANT LINFORD , HODSON DAVID , DISTASO WALTER , NINOV NIKOLAY N. , RUTTER GUY A.
Published: June 2019

Aim: In-vitro studies reveal a subpopulation of β cells which co-ordinate insulin release from pancreatic islets. This connectivity has not been established in 3 dimensions across the islet or in the living animal. We investigated this in the living zebrafish D. rario or in mouse islets engrafted in the anterior eye chamber (ACE), using the recombinant Ca2+ sensor GCaMP6 under the insulin promoter/C57Bl6, Ins1Cre:GCaM6ff/f.

Methods: Whole islet live imaging of D. rario primary islets in situ at an acquisition rate of 0.8Hz, covering 700μm, was achieved using resonant scanner technology with a Leica inverted laser scanning confocal system, using a 25X/N.A. 0.95 water lens. Fully vascularized islets in the murine ACE were imaged under isofluorane anaesthesia on a modified Nikon Ti-E spinning disc confocal microscope (20x 0.75 NA water immersion objective, 3 Hz frame rate).

Results: 3-dimensional β cell connectivity (Pearson R statistic) significantly rose to 88% across the zebrafish islet in response to glucose stimulation. Focused ablation of the co-ordinating “hub” (but not follower) β cells significantly attenuated the subsequent response to a glucose challenge. In mouse islets, high circulating glucose levels were also associated with a significant rise in 3D beta cell connectivity from 65 to 86% (n=3; p=0.02). Granger causality analysis on these islets revealed that even on prolonged (>10 minute) glucose stimulation, pan-islet connectivity was sustained and the most highly connected cells were those in the region of the islet from which Ca2+ waves emanated. Binarized signal analysis (n=6) separately confirmed the presence of a highly-connected β cell subpopulation (8.7 ± 3.6% of cells).

Conclusions: These studies evidence beta cell “hubs” which are responsible for the initiation and sustained coordination of the Ca2+ response to glucose across fish and mouse islets in vivo.

Disclosure

V. Salem: None. L.F. Delgadillo Silva: None. K. Suba: None. A. Martin Alonso: None. W. Kim: None. V. Kalogianni: None. N. Akhtar: None. N. Mousavy Gharavy: None. E. Georgiadou: None. I. Leclerc: None. L. Briant: None. D. Hodson: None. W. Distaso: None. N.N. Ninov: None. G.A. Rutter: Consultant; Self; Sun Pharma.

Funding

Diabetes UK; UK Wellcome Trust
425-P: Inhibition of miR-181c Rescues Diabetes-Impaired Angiogenesis
Authors: SOLLY EMMA , HOURIGAN SAMUEL T. , MULANGALA JOCELYNE , PSALTIS PETER J. , BARTOLO BELINDA DI , NG MARTIN , NICHOLLS STEPHEN , BURSILL CHRISTINA , TAN JOANNE T.M.
Published: June 2019

Diabetic vascular complications are characterized by impaired angiogenic responses to ischemia, resulting in delayed healing following tissue injury. MicroRNAs (miRNAs), small non-coding RNAs that target gene expression, regulate angiogenesis in a time-dependent manner. Using a miRNA array approach, we discovered that miR-181c was regulated by angiogenic stimuli but had no previous reported role in diabetes. We sought to elucidate the role of miR-181c in diabetes-impaired angiogenesis. We found that miR-181c was inhibited in endothelial cells (ECs) exposed to hypoxia but were, conversely, elevated in high glucose. To assess the importance of miR-181c in angiogenesis, ECs were transfected with mimics or antimiRs to mimic or suppress miR-181c activity respectively. Cells with increased miR-181c activity had a reduced capacity to form tubules and lower VEGFA protein levels. Conversely, suppression of miR-181c activity augmented tubule formation and VEGFA expression. This suggests that miR-181c plays an inhibitory role in angiogenesis, at least in part, through VEGFA regulation. In vivo, miR-181c was temporally tracked following ischemic induction in the hindlimb. Early post-ischemia, when angiogenesis is important, diabetic mice had elevated miR-181c levels compared to nondiabetic controls. However, at later timepoints, miR-181c levels dropped to levels similar to nondiabetic mice. This indicates that miR-181c suppression may be critical to facilitate early responses to ischemia. Diabetic mice were injected with antimiR-181c or antimiR-Neg to determine if miR-181c knockdown rescues diabetes-impaired angiogenesis. Inhibition of miR-181c completely rescued diabetes-impaired blood flow reperfusion, restoring it back to that of nondiabetic levels. Taken together, this study has identified that miR-181c plays a key role in the regulation of angiogenesis in ischemia and in diabetes. This highlights a potential new therapeutic approach for the treatment of diabetic vascular complications.

Disclosure

E. Solly: None. S.T. Hourigan: None. J. Mulangala: None. P.J. Psaltis: None. B. Di Bartolo: None. M. Ng: None. S. Nicholls: Consultant; Self; Amgen Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Cerenis, CSL Behring, Eli Lilly and Company, Merck Sharp & Dohme Corp., Novo Nordisk Inc., Omthera, Pfizer Inc., Resverlogix Corp., Roche Pharma, Sanofi, Takeda Pharmaceutical Company Limited. Research Support; Self; Amgen Inc., AstraZeneca, Athernova, Cerenis, Eli Lilly and Company, InfraReDx, Novartis Pharmaceuticals Corporation, Resverlogix Corp., Sanofi. C. Bursill: None. J.T.M. Tan: None.

Funding

Diabetes Australia
426-P: ROCK2 Regulates Monocytic Migration and Cell-to-Cell Adhesion in Vascular Endothelial Cells
Authors: TAKEDA YUSUKE , MATOBA KEIICHIRO , KAWANAMI DAIJI , NAGAI YOSUKE , UTSUNOMIYA KAZUNORI
Published: June 2019

A pro-inflammatory lipid mediator, lysophosphatidic acid (LPA), is a potent activator of the RhoA/Rho-kinase signaling pathway and has been shown to induce the expression of chemokines and cell adhesion molecules (CAMs). LPA levels are elevated in the circulation of patients with acute coronary syndrome and have also been detected inside human atherosclerotic specimens. However, the involvement of Rho-kinase isoforms (i.e., ROCK1, ROCK2) in LPA-mediated expression of chemokines and CAMs in endothelial cells is not fully understood. In this study, we investigated isoform-specific roles of Rho-kinase in LPA-mediated MCP-1 and E-selectin expression using human aortic endothelial cells (HAEC). Exposure of HAEC to LPA resulted in an increase of MCP-1 and E-selectin expression at both mRNA and protein levels. Rho-kinase inhibitor abolished MCP-1 and E-selectin protein expression, indicating that LPA induces MCP-1 and E-selectin expression via Rho-kinase activation. From a mechanistic standpoint, phosphorylation of IκBα, a well-characterized initial step in NF-κB activation, was decreased by Rho-kinase inhibition in the LPA-stimulated cells. Furthermore, using isoform specific siRNAs, we found that Rho-kinase isoform ROCK2 is the key mediator of LPA-induced MCP-1 and E-selectin expression. Consistently, ROCK2 but not ROCK1 controlled LPA-induced monocytic migration and adhesion in functional assays using HAECs and human monocytic cells (THP-1). In the present study, we demonstrate that endothelial ROCK2 mediates monocyte migration and cell adhesion induced by lipid mediator. In order to further explore endothelial-specific role of ROCK2 in vivo, we generated endothelial ROCK2 knockout mouse (EC-ROCK2KO) by mating of ROCK2 flox mice with VE-cadherin-cre mice. Further investigation using HFD (high-fat diet) mouse model is ongoing. ROCK2 could be an important therapeutic target against atherosclerosis.

Disclosure

Y. Takeda: None. K. Matoba: Research Support; Self; Daiichi Sankyo Company, Limited, Novo Nordisk Inc., Sanofi, Shionogi & Co., Ltd., Takeda Pharmaceutical Company Limited. D. Kawanami: Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc. Y. Nagai: None. K. Utsunomiya: None.

Funding

Japan Society for the Promotion of Science; Uehara Memorial Foundation; MSD Life Science Foundation
429-P: Therapeutic Inertia: Gap between Goals and Achievement between the T1DX and DPV Registries
Authors: MAAHS DAVID M. , HERMANN JULIA , FOSTER NICOLE C. , KARGES WOLFGANG , MILLER KELLEE , DOST AXEL , WEINSTOCK RUTH S. , PAVEL MARIANNE , SHAH VIRAL N. , HOLL REINHARD W.
Published: June 2019

Despite increased cardiovascular (CV) mortality in people with type 1 diabetes (T1D), treatment of CV risk factors may not be optimized. International registry comparisons highlight opportunities to improve clinical care and improve outcomes. We analyzed data from 28,494 patients from the German/Austrian DPV registry and 19,442 patients from the U.S. T1D Exchange (T1DX) Clinic registry with data between 01/2016 and 03/2018. Percent of patients by age category with HbA1c <7.5%, hypertension (HTN) and its treatment, dyslipidemia and its treatment, urine albumin to creatinine ratio >30 mg/g, BMI >30 kg/m2, and current smoking are reported. Logistic regression models by age group adjusted for sex and diabetes duration were used to calculate whether registry differences existed. The prevalence of obesity is lower in the DPV than the T1DX registry (Table). In those age <26 years more DPV than T1DX patients had A1c <7.5%. Hypertension was more prevalent in the DPV for those <26 years, but less prevalent for those >50 years. Dyslipidemia was more prevalent in DPV for those <50 years, but less for those <50 years. Elevated ACR and smoking are higher and HTN and dyslipidemia treatment are lower in the DPV than in the T1DX. Vascular disease remains the leading cause of morbidity and mortality in T1D. Comparison of risk factors and their treatment in these registries highlights opportunities to improve care and outcomes.

Disclosure

D.M. Maahs: Advisory Panel; Self; Novo Nordisk Inc. Consultant; Self; Abbott, Sanofi. Research Support; Self; Dexcom, Inc., Tandem Diabetes Care. J. Hermann: None. N.C. Foster: None. W. Karges: None. K. Miller: None. A. Dost: None. R.S. Weinstock: Consultant; Self; Insulogic LLC. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Jaeb Center for Health Research, Kowa Pharmaceutical Europe Co. Ltd., Medtronic MiniMed, Inc., Oramed Pharmaceuticals. Other Relationship; Self; Unitio, Inc. M. Pavel: None. V.N. Shah: Advisory Panel; Self; Sanofi US. Consultant; Self; Dexcom, Inc. R.W. Holl: None.

Funding

The Leona M. and Harry B. Helmsley Charitable Trust; German Center for Diabetes Research; German Diabetes Association; European Foundation for the Study of Diabetes; INNODIA
437-P: Association of Skeletal Muscle Mass and Grip Strength with Cardiovascular Disease in Korean Patients with Type 2 Diabetes
Authors: SHIN JANG YEL
Published: June 2019

Sarcopenia is a progressive decline in muscle mass and strength with aging. Few studies have investigated that lower muscle mass or strength were associated with cardiovascular disease (CVD). We evaluated whether skeletal muscle mass and grip strength were associated with CVD in type 2 diabetic patients. A total of 1,038 subjects with type 2 diabetes were recruited. We measured appendicular skeletal muscle mass (ASM) using dual energy X-ray absorptiometry and used ASM/BMI for muscle mass quantity. Grip strength (GS) was assessed using a JAMA dynamometer. Patients were divided into sex-specific three groups of ASM/BMI and GS. CVD included coronary heart disease (CHD), cerebral infarction (CI), and peripheral artery disease (PAD) diagnosed by ICD-10. Patients with CVD, CHD, CI, and PAD were 232 (22.4%), 151 (14.5%), 68 (6.6%), and 38 (3.7%), respectively. Subjects with lowest tertile of ASM/BMI were more likely to be elderly and hypertensive, had higher values of systolic BP (SBP), waist circumference (WC), BMI, diabetes duration (DD), C-reactive protein (CRP), serum creatinine (Cr), urine albumin excretion (UAE), and HbA1c, and had lower values of diastolic BP (DBP) and HDL compared with those with highest. Whereas, participants with lowest tertile of GS had higher values of age, DD, Cr, and UAE and had lower values of BMI, DBP, HDL, and LDL than those with highest. The prevalence of CVD, CHD, CI, and PAD with decreasing tertile of ASM/BMI and GS increased proportionally (p<0.001). Odd ratios (ORs) for presence of CVD after adjusted for age with decreasing tretile of ASM/BMI and GS were significantly increased: OR (95% CI) = 1.93 (1.23-3.03) and 2.01 (1.28-3.16) in middle vs. 3.35 (2.16-5.20) and 2.87 (1.80-4.57) in lowest. ORs were maintained significantly after further adjusted for hypertension, current smoking, DD, WC, BMI, BP, HDL, CRP, Cr, HbA1c, and UAE. We found that lower muscle mass and strength were independently associated with CVD in patients with type 2 diabetes.

Disclosure

J. Shin: None.
44-LB: Hypothalamic Glucocorticoid Action Regulates Lipid Metabolism
Authors: CARDOSO MIGUEL , WANG MARK , LING EMILY , LEE HONGJU (JENNIFER) , BEAULIEU-BAYNE EMILIE , YUE JESSICA T.
Published: June 2019

Aberrant lipid homeostasis is a feature of diabetes and obesity. This metabolic imbalance is associated with excessive levels and/or actions of glucocorticoids (GCs) and contributes to dyslipidemia including elevated hepatic triglyceride (TG)-rich very low-density lipoprotein (VLDL-TG) secretion. The mediobasal hypothalamus (MBH) is a nutrient and hormone sensing brain region that regulates whole-body lipid metabolism and VLDL-TG secretion. We aimed to assess GC action in the MBH in regulating VLDL-TG secretion in health and diet-induced hyperlipidemia.

Sprague Dawley rats received stereotaxic MBH cannulation and vascular catheterizations to enable direct MBH infusions, intravenous injections, and blood sampling. Plasma TG levels and the rate VLDL-TG secretion were measured in 10h-fasted rats following intravenous poloxamer injection with concomitant MBH infusions. In healthy rats, direct MBH GC infusion increased VLDL-TG secretion compared to controls. MBH GC action is mediated via its receptor (GR) since mifepristone (GR antagonist) co-infusion, chronic inhibition of GRs in the MBH with GR shRNA, or inhibition of heat shock protein 90 (required for in vivo GR function) negated the MBH GC-induced increases in VLDL-TG secretion rate. High-fat diet-fed rats had both elevated basal plasma GC levels and hepatic TG secretion compared to healthy rats. Remarkably, chronic MBH GR loss-of-function lowered VLDL-TG secretion in high-fat diet-induced hyperlipidemic rats, independent of changes in body weight.

Here, we provide novel in vivo evidence that MBH GC action modulates VLDL-TG secretion and suggest that blocking excessive GC action in the brain improves lipid homeostasis in diet-induced hyperlipidemic rats. These findings may provide insight on lowering lipid levels in diabetes and obesity.

Disclosure

M. Cardoso: None. M. Wang: None. E. Ling: None. H. Lee: None. E. Beaulieu-Bayne: None. J.T. Yue: None.

Funding

Diabetes Canada; Natural Sciences and Engineering Research Council of Canada; Canadian Lipoprotein Conference