bluuebunny/crossref_metadata_2025 huggingface dataset:
Acute Caloric Restriction Leads to Loss of Hypoglycemic Counter-Regulation in Mice following Short-Term Refeeding
Authors: MCDOUGAL DAVID
Published: June 2018

Background: Hypoglycemia-associated autonomic failure (HAAF) is a maladaptive failure in glucose counter-regulation known to be caused by recurrent exposure to insulin-induced hypoglycemia. Fasting or caloric restriction is the only natural physiological process which induces prolonged moderate-to-severe hypoglycemia similar to the antecedent of HAAF. In this study we tested the hypothesis that exposure to fasting-induced hypoglycemia can cause HAAF-like symptoms in mice.

Methods: Two groups of mice were used, an ad libitum (ad lib) fed group (n=6) and a caloric restriction (CR) group (n=6). CR mice were placed on 60% caloric restriction for 6 consecutive days. Ad lib mice were housed in an identical manner but fed ad libitum during this same period. Following 6 days of restriction, CR mice were given ad lib access to food for 16 h. After the 16 h period of refeeding, both CR and ad lib mice began a 6 h fast which was immediately followed by a hypoglycemic insulin tolerance test (ITT). ITTs consisted of a variable dose of insulin (1.4-1.75 U/kg, IP) which produced similar levels of hypoglycemia in each group (∼45 mg/dL).

Results: As expected, mice exposed to 6 days of 60% CR lost ≈30% of their initial body weight, and experienced a gradual fall in blood glucose to an average minimum of 67.9 mg/dL. CR mice displayed a significantly reduced fasting glucose level relative to ad lib mice (102.7 and 171.0 mg/dL respectively, p = 0.0002) as well as reduced serum glucagon levels in response to the ITT (6.0 and 30.4 pg/mL respectively, p = 0.0182).

Conclusions: Our results demonstrate that exposure to fasting-induced hypoglycemia produces HAAF-like symptoms in mice following refeeding. These results suggest that HAAF may be a result of an adaptive response to fasting which is inappropriately elicited during exposure to insulin-induced hypoglycemia. Further characterization of the effects of CR on the induction of deficits in glucose counterregulation warrants further investigation.

Disclosure

D. McDougal: None.
Duodeno-Jejunal Bypass Liner for the Treatment of Diabetes Mellitus in Obese Patients—Completeness of Duodenal Blinding as the Key Factor for Efficacy
Authors: BENES MAREK , DRASTICH PAVEL , HUCL TOMAS
Published: June 2018

Introduction: The global increase in obesity incidence results in an increase of type 2 diabetes mellitus (T2DM). Surgical treatment has proven to be effective, however it carries a high risk of complications. The duodenal-jejunal bypass liner (EndoBarrier®, GI Dynamics, EB) is an endoscopic implant that mimics the intestinal bypass portion of the Roux-en-Y Gastric Bypass. It results in weight loss and improvements in glucose control in obese patients with T2 diabetes mellitus (T2DM).

Aims and Methods: This is a analysis of a prospective, controlled, multicentre study aimed to identify factors associated with an outcome of EB for T2DM.

Results: Seventy subjects (45 with an implant, 25 controls) were included in the study. The groups were comparable with respect to age, gender, BMI (mean 41.7 vs. 39.5 kg/m2), T2DM duration (7.8 vs. 8.3 years), HbA1c level (88 vs. 86 mmol/mol) and T2DM treatment. In the EB group, all devices were successfully implanted. Only 6 devices had to be explanted prior to the end of the 10 months study period (bleeding, dislocation and need for ERCP because of choledocholithiasis). At 10 months there was significantly greater weight loss and %EWL (19% vs. 7% and 43 vs. 12) and significantly improved long term compensation of T2DM marker HbA1c (decreased by 25 vs. 10 mmol/mol) in the EB group. T2DM medicinal treatment could be reduced in more device subjects than controls. There was no serious adverse event. Deepness of anchor ingrow, lower initial BMI and lower body height were identified as positive factors for efficacy of EB for T2DM compensation.

Conclusion: The EB is safe when implanted for 10 months, and results in significant weight loss and HbA1c reduction. Deepness of anchor ingrow, lower initial BMI and lower body height could be positive factors for efficacy.

Disclosure

M. Benes: None. P. Drastich: None. T. Hucl: None.
Elevated Insulin Clearance Contributes to the Reduced Peripheral Insulin Concentration Observed in Obese Young Asian with Type 2 Diabetes
Authors: PACINI GIOVANNI , TURA ANDREA , HOR CHEE PENG , LIM SHUEH LIN , TAN FLORENCE , TONG CHIN VOON , HONG JANET Y.H. , ZAIN FUZIAH MD , HOLST JENS J. , MOHAMUD WAN NAZAIMOON , YEOW TOH PENG
Published: June 2018

Introduction: The importance of insulin clearance (CLins) on insulin dynamics and the role exerted in controlling glucose in type 2 subjects has recently been appreciated. Moreover, evidence in mice showed that CLins is reduced by incretin hormones at physiological levels.

Aim: To evaluate in 29 young (19±1 years) multi-ethnic Asian type 2 subjects (T2D) the contribution of CLins to insulin dynamics and if an association exists with GLP-1.

Methods: T2D (15F/14M; BMI=30.2±0.9 kg/m2; HbA1c=8.8±0.4% [73±4.4 mmol/mol]; mean duration of disease 2.5 years) had fasting glucose Gb=192±14 mg/mL and insulin Ib=19±3 µU/mL. They were compared to 17 (8F/9M) age-matched nondiabetic overweight (BMI=27.8±1.0, p=0.1) control subjects (CT, Gb=82±1, p<0.0001; Ib=12±2, p=0.09). All underwent a 75g 2h OGTT with measurements of glucose, insulin, C-peptide and GLP-1. Insulin secretion (ISR) was assessed by deconvolution from peripheral C-peptide; CLins) by ISR/AUCinsulin; beta cell function (BCF) by AUCC-peptide/AUCglucose; insulin sensitivity by OGIS model. GLP-1 effect on BCF (GLPE) was empirically calculated as BCF/AUCGLP-1.

Results: T2D were insulin resistant (OGIS=277±16 mL/min m2 vs. 424±14) and had low BCF (82±11 pmolC-peptide/mmolglucose vs. 123±10), both p<0.0001. Lower total AUCinsulin was observed in T2D (4.7±0.7 U/L 2h vs. 11.0±1.4, p<0.0001), with higher CLins (2.06±0.19 L/min) than in CT (1.46±0.10, p=0.023). CLins directly correlated with OGIS in CT (R=0.55, p=0.02), but not in T2D (p=0.98). Despite no difference was observed in AUCGLP-1 (1.6±0.1 nmol/L 2h vs. 1.7±0.1 in CT, p=0.63), which did not correlate with CLins (p>0.2), GLPE was lower in T2D (6.2±0.9 vs. 23.4±2.3, p<0.0001).

Conclusion: Reduced peripheral insulin availability in obese young Asian T2D is due not only to a markedly impaired beta cell function and reduced GLP-1 effect, but also to an increased insulin clearance, which however shows no association to GLP-1 response.

Disclosure

G. Pacini: None. A. Tura: None. C. Hor: None. S. Lim: None. F. Tan: None. C. Tong: None. J.Y.H. Hong: None. F. Md Zain: None. J.J. Holst: Advisory Panel; Self; Novo Nordisk A/S. Board Member; Self; Zealand Pharma A/S. Speaker's Bureau; Self; Merck Sharp & Dohme Corp., AstraZeneca. Research Support; Self; Danish Diabetes Academy, Novo Nordisk Foundation. Other Relationship; Self; Antag Therapeutics. Other Relationship; Spouse/Partner; Antag Therapeutics. W. Mohamud: None. T. Yeow: None.
Endobarrier (EB) in Type 2 Diabetes/Prediabetes with Obstructive Sleep Apnea Study—Preliminary Results
Authors: YADAGIRI MAHENDER , KINNEY FIONA Y. , ASHMAN NATALIE , LANG MICHAEL H. , FOGDEN EDWARD , ANDERSON MARK , BLEASDALE JOHN , WALTON CHRISTOPHER , GREENSTONE MICHAEL A. , RYDER ROBERT E.
Published: June 2018

Aims: To assess the extent to which patients with type 2 diabetes or prediabetes, obesity (BMI 30-45kg/m²) and moderate OSA requiring continuous positive airway pressure ventilation (CPAP) are able to discontinue CPAP following EB-related weight loss (ISRCTN33788132).

Methods: We assessed Apnoea Hypopnoea Index (AHI), weight and HbA1c before and following EB. The full study involves 18 patients receiving EB for up to 1 year with 1 year follow-up.

Results: We report here the 1st 11 patients {9/11 (82%) female, 9/11 (82%) type 2 diabetes, 2/11 (18%) prediabetes, mean ± SD age 52.6 ± 9.7 years} to reach at least 3 months EB treatment. In 3 months, weight fell by 6.9 ± 3.3kg from 103.7 ± 14.8 to 96.8 ± 14.8kg (p<0.001), mean BMI by 2.5 ± 1.3kg/m² from 37.1 ± 3.6 to 34.6 ± 3.7kg/m² (p<0.001), mean HbA1c by 0.9±1.0% from 8.0±3.8 to 7.1±3.7% (p=0.025). OSA improved (Figure). Prior to EB, all 11 patients had AHI in moderate sleep apnoea range (15-29.9 events/hour). Following EB, the AHI of 7/11(64%) patients fell below the moderate sleep apnoea threshold of 15 events/hour, such that they no longer required CPAP. Of the remaining 4 patients, 1 came off CPAP at 6 months.

Conclusion: These preliminary results are encouraging in that EB has already allowed 8/11(73%) patients to discontinue CPAP, in addition to glycaemic and weight benefits. Discontinuing CPAP is beneficial to health services but especially to patients.

Disclosure

M. Yadagiri: None. F.Y. Kinney: None. N. Ashman: None. M.H. Lang: None. E. Fogden: None. M. Anderson: None. J. Bleasdale: None. C. Walton: None. M.A. Greenstone: None. R.E. Ryder: Other Relationship; Self; AstraZeneca. Speaker's Bureau; Self; Bioquest, Janssen Pharmaceuticals, Inc.. Other Relationship; Self; Janssen Pharmaceuticals, Inc.. Advisory Panel; Self; Novo Nordisk A/S.
Incretin and Cardiovascular Effects of Weight Loss and Remission of Prediabetes
Authors: STENTZ FRANKIE B. , AMMONS ANN
Published: June 2018

Since the incretins GLP-1 and GIP have important roles in insulin sensitivity and have been shown to be effective in pharmacological treatment of type 2 Diabetes, we studied these incretin level changes with weight loss and remission of prediabetes in 24 obese women and men with prediabetes. Our studies have shown that prediabetes subjects on a 6 month HP diet (30% protein, 30% fat, 40% CHO) had 100% remission of prediabetes compared to only 33% on a HC diet (15% protein, 30%fat, 55% CHO). Subjects on the HP diet had increased satiety compared to the HC diet, therefore, we studied HP and HC diet effects on ghrelin levels. Additionally, since cardiovascular risk factors (CVRF) decreased more in the HP than the HC diet, we determined if the B-Type Natriuretic Peptide(BNP) released from the heart was affected by either diet. GLP-1, GIP, Ghrelin, and BNP levels were determined with a Meal Tolerance Test (MTT) at baseline and after 6 mo on HP and HC diets where all food was provided.

The active GLP-1 and total GIP area under the curves for the HP MTT increased significantly from baseline vs. 6 months (7860±31 vs. 8970±37 pmol/l/min, p=0.001) and (21,350±63 vs. 27,240±72 pg/ml/min, p=0.001) respectively. The ghrelin decreased significantly from baseline vs. 6 months on the HP MTT (338±21 vs. 78±14 pg/ml/min, p= 0.005). BNP decreased significantly from baseline (126±11pmol/l) to 6 months (78±9 pmol/l), p=0.01. HP diet had a significantly greater increase in GLP-1 and GIP than the HC diet. HP ghrelin results demonstrate that HP diet can induce satiety and is more effective than the HC diet. The BNP decrease in both groups demonstrates the improvement in heart tissue with the HP diet having a greater effect than the HC diet.

Although weight loss was similar (9.8% in HP vs. 11.3% in HC, p=0.692), this study demonstrates that the HP diet increases GLP-1 and GIP which may be responsible in part for the improved insulin sensitivity, and decreases BNP with greater improvement in CVRF with remission of prediabetes.

Disclosure

F.B. Stentz: None. A. Ammons: None.
Effects of Laparoscopic Sleeve Gastrectomy on Insulin Secretion Patterns in Young Obese Patients with Acanthosis Nigricans
Authors: QIAN CHUNHUA , QU SHEN , ZHU CUILING , GAO JINGYANG , MEI FANGYUN , BU LE
Published: June 2018

Background: Acanthosis nigricans (AN) is a skin condition commonly often accompanied by insulin resistance and hyperinsulinemia. Patients with AN are prone to diabetes, especially in young obesity. Metformin were usually used to treat these patients. However, reports about the effectiveness of LSG surgery on young obese patients with AN are rare.

Objective: The aim of our study was to evaluate the efficacy of LSG surgery on insulin secretion patterns and anthropometric variables for obese patients with AN.

Methods: 30 young obese patients with AN were recruited for this study. Before and at 12 months after the LSG surgery, BMI and blood samples were collected. OGTT were used to evaluate the insulin level and its rhythm of secretion. AN extent and AN texture were determined according to the scale developed and validated.

Results: For these patients, BMI decreased from 40.98±4.93 kg/m2 to 27.27±4.13 kg/m2 at 12 months after surgery. Along with weight loss, triglycerides and total cholesterol decreased significantly, and high-density lipoprotein cholesterol (HDL-C) increased significantly as of 12 months after surgery. But the change of low-density lipoprotein cholesterol (LDL-C) is not obvious. A significant decrease in insulin resistant (HOMA-IR) was observed. The insulin release curves showed hyperinsulinemia and abnormal rhythm of secretion before surgery. At 12 months after surgery the fasting, 2h postprandial insulin levels and the rhythm of insulin secretion were restored to normal level. But the insulin levels at 30min and 60min were higher than before surgery. The extent of AN for most patients were improved. Only 23.4% patients still were severe (vs. 59.3% before surgery). More than 50% patients were remission.

Conclusion: In this study of LSG in obesity patients with AN, we found significant improvements in weight, dyslipidemia, hyperinsulinemia and AN extent. LSG will be a new effective therapeutic approach for young obesity patients with AN.

Disclosure

C. Qian: None. S. Qu: None. C. Zhu: None. J. Gao: None. F. Mei: None. L. Bu: None.
Influence of Race, Ethnicity, and Behavioral Factors on Childhood Obesity
Authors: AYINE PRISCILLA , PARRA EMILY P. , JEGANATHAN RAMESH B. , THANGIAH GEETHA
Published: June 2018

Childhood obesity is a public health concern resulting from a variety of reasons. In the United States, approximately 12.7 million children and adolescents are classified as obese. 23.8% of African American children are obese compared with 13.1% of whites between ages 6 to 11 years. In Alabama, 35.5% children are overweight and obese; it is 6th highest ranked in United States. The most common causes of childhood obesity are behavioral, genetic factors, or a combination of both.

In this study we investigated the differences in some of the behavioral factors such as sleeping time, dinner time and television watching time in racial ethnic groups. 24 white/European American (EA) and 30 black/African American (AA) elementary school children aged between 6 to 10 years participated in this study. Height and weight were measured, without shoes and wearing only light clothing. Body mass index (BMI) percentile was calculated using Centers for Disease Control and Prevention growth reference. Parents reported child’s typical weekday bedtime, dinner time and television watching time of their children.

35% of EA children slept later than 8:30 pm during school days compared to 68% of AA children. Only 4% of EA children had late dinner (after 7 pm), whereas 50% of AA children had late dinner during week days. 21% of EA children watched television for more than 1 hour in comparison to 79% of AA children every day. Our findings suggest that insufficient sleep, late dinner schedule, and longer television exposure are some of the behavioral factors that contribute to greater percentage of childhood obesity in AA than EA children.

Disclosure

P. Ayine: None. E.P. Parra: None. R.B. Jeganathan: None. G. Thangiah: None.
Real-World Data from the MiniMed™ 670G System Commercial Launch—Patients Previously Using MDI Therapy
Authors: GOPALAKRISHNAN SHWETA , AGRAWAL PRATIK , STONE MICHAEL , FOGEL CATHERINE , LEE SCOTT W.
Published: June 2018

The Medtronic MiniMed™ 670G hybrid closed-loop system with SmartGuard™ technology was released for commercial use in the U.S. in March 2017. Data from patients previously managing their diabetes with multiple daily insulin injections (MDI) therapy who transitioned to the MiniMed™ 670G system are reported. MiniMed™ 670G system data from patients who voluntarily uploaded to CareLink™ Personal software, from 3/17/17 to 12/14/17 (n=158), were de-identified and analyzed if there were ≥7 days of sensor wear. Comparisons were made of glycemic metrics between 4,529 patient-days in the initial open-loop Manual Mode and 14,1patient-days after initiating closed-loop Auto Mode features. The Table shows glycemic metrics during Manual Mode and after Auto Mode start and that time spent in target range increased, while time spent in low and high glucose levels decreased. Real-world findings from patients previously using MDI who transitioned to the MiniMed™ 670G system demonstrated increased TIR and reduced time in hypoglycemia and hyperglycemia, consistent with pivotal trial1 results in patients previously using pump therapy +/- CGM. Prior MDI patients also showed greater TIR the more time spent in Auto Mode.

Disclosure

S. Gopalakrishnan: Employee; Self; Medtronic. P. Agrawal: None. M. Stone: None. C. Fogel: Employee; Self; Medtronic. S.W. Lee: Employee; Self; Medtronic.
Energy Expenditure during Sleep Is an Independent Marker of Insulin-Resistance—A Cross-Sectional Study of Overweight/Obese Subjects
Authors: TUCCINARDI DARIO , MADDALONI ERNESTO , CAPASSO ILARIA , RAMPA LORENZO , SOARE ANDREEA , MAGGI DARIA , DEFEUDIS GIUSEPPE , KYANVASH SHADI , MAURIZI ANNA RITA , POZZILLI PAOLO , MANFRINI SILVIA
Published: June 2018

Normal sleep is characterized by increased glycogen stores and reduced glucose turnover by metabolically active tissues, thus it is a state of decreased metabolism. It has been reported that reduced sleep quality is associated with increased insulin-resistance and disrupted metabolism. However, the relationship between energy expenditure during sleep (EEDS) and insulin-resistance (homeostasis model assessment, HOMA-IR) has not been clarified. The aim of this study was to investigate the role of EEDS in the association between sleep quality and insulin resistance. A total of 226 individuals [women 72.1%; mean (DS) age 50.46 (±13.1) years; BMI 33.4 (±8.41) Kg / m2] undergoing the oral glucose tolerance test (OGTT) were also asked to wear a metabolic holter (Armband) for 3-days, to assess their daily energy expenditure. The Armband is capable of measuring energy expenditure every minute, but also the duration and quality of sleep. The EEDS corrected for weight (EEDS/WEIGHT) was lower in patients with insulin resistance, HOMA-IR above 2.5 (p <.001). The EEDS/WEIGHT was inversely related to HOMA-IR (r = -0.23; p <.001) and directly to the quality of sleep (r = 0.15; p <.001). Using a multivariate linear regression, with sleep quality, age, gender, glucose and insulin area under the curve (AUC) and EEDS/ WEIGHT as independent variables, the HOMA-IR value was independently and significantly predicted by the insulin AUC (p <.001, beta = 0.538), by gender (p <.01, beta = 0.155) and by the EEDS/WEIGHT (p <.004, beta = -0.173). EEDS is an independent marker of insulin resistance. The mechanism underlying this correlation may be explained by the state of reduced glucose uptake during sleep, which it is further increased during insulin-resistance conditions, with a consequential reduction in energy production. This finding may provide another explanation to the inverse association between sleep quality and metabolic disorders such as type 2 diabetes and obesity.

Disclosure

D. Tuccinardi: None. E. Maddaloni: Speaker's Bureau; Self; Merck & Co., Inc.. Research Support; Self; European Foundation for the Study of Diabetes. I. Capasso: None. L. Rampa: None. A. Soare: None. D. Maggi: None. G. Defeudis: None. S. Kyanvash: None. A. Maurizi: None. P. Pozzilli: Research Support; Self; Sanofi. Speaker's Bureau; Self; Eli Lilly and Company. Research Support; Self; Merck Sharp & Dohme Corp.. S. Manfrini: None.
First Risk–Benefit Data from the Worldwide Endobarrier Registry
Authors: RYDER ROBERT E.J. , MUNRO LYNNE , MCMASTER JESSICA J. , BESSELL JUSTIN , BASCOMB JEANINE M. , COLLINS JANE E. , KOW LILIAN , CHISHOLM JACOB , SOURIJ HARALD , PFERSCHY PETER N. , TEARE JULIAN P. , MASON JOHN C. , BYRNE JAMES P. , WYRES MELANIE C. , CULL MELISSA L. , BURBRIDGE WYN , IRWIN SUSAN P. , YADAGIRI MAHENDER , FOGDEN EDWARD , ANDERSON MARK , GUPTA PIYA SEN , BENES MAREK
Published: June 2018

Uncertainty exists about risk:benefit of proximal intestinal exclusion with Endobarrier (EB), a novel endoscopic duodenal jejunal liner device for obesity, both with and without diabetes. In view of this, during 2017, an independent, secure, on-line registry was established under the auspices of the Association of British Clinical Diabetologists, for the collection of safety and efficacy data worldwide. As of December 2017, data had been entered on 403 patients {age 51.3 ± 11.8 year, 62% male, 89% europid, 74% diabetes, BMI 42.6 ± 10.2 kg/m2} from 13 centres in 4 countries: Australia, Austria, Czech Republic and United Kingdom. EB led to many benefits, including: in those with both baseline and explant data, mean ± SD weight fell by 14.5 ± 10.3 kg from 125.3 ± 26.7 to 110.8 ± 26.4 kg (n = 265 p<0.001), HbA1c by 1.4 ± 1.6%, from 8.7 ± 1.8 to 7.2 ± 1.2% (n = 195, p<0.001) and systolic BP fell from 138.5 ± 18.1 to 130.0 ± 17.2 mmHg (n = 149, <0.001). There were 23 (5.7%) serious adverse events (SAE) and 37 (9.2%) less serious AEs (Table). All SAE patients made a full recovery. The median (range) weight loss in those with early removal for GI bleed was 6.5 (0-29) and with early removal for liver abscess was 17.2 (7-21) kg. Some SAEs could have been avoided if patients had adhered to guidelines. The benefits of EB therapy are likely to reduce the complications of diabetes. This 1st international data from the EB registry suggests that the likely benefits of EB, far outweigh the risks.

Disclosure

R.E.J. Ryder: Other Relationship; Self; AstraZeneca. Speaker's Bureau; Self; Bioquest, Janssen Pharmaceuticals, Inc.. Other Relationship; Self; Janssen Pharmaceuticals, Inc.. Advisory Panel; Self; Novo Nordisk A/S. L. Munro: None. J.J. McMaster: None. J. Bessell: None. J.M. Bascomb: None. J.E. Collins: None. L. Kow: None. J. Chisholm: None. H. Sourij: Speaker's Bureau; Self; Boehringer Ingelheim GmbH, Novo Nordisk A/S, Amgen Inc., Sanofi, MSD K.K.. Research Support; Self; AstraZeneca, Boehringer Ingelheim GmbH, MSD K.K., GI Dynamics Inc.. P.N. Pferschy: None. J.P. Teare: Research Support; Self; GI Dynamics Inc.. J.C. Mason: None. J.P. Byrne: None. M.C. Wyres: None. M.L. Cull: None. W. Burbridge: Other Relationship; Self; Menarini Group. S.P. Irwin: None. M. Yadagiri: None. E. Fogden: None. M. Anderson: None. P. Sen Gupta: None. M. Benes: None.
Polygenic Risk Testing at the Time of Diagnosis of Diabetes Could Help Prevent Its Micro- and Macrovascular Complications
Authors: HAMET PAVEL , HARVEY FRANCOIS , HALOUI MOUNSIF , WOODWARD MARK , TAHIR RAMZAN , MAROIS-BLANCHET FRANÇOIS-CHRISTOPH , LONG CAROLE , MARRE MICHEL , HARRAP STEPHEN , CHALMERS JOHN , TREMBLAY JOHANNE
Published: June 2018

Cardiovascular and renal complications of T2D represent a human and societal burden. Early detection of susceptibility to complications could help their prevention. Clinical scores are useful to predict hard outcomes but genomic score can be applied earlier than a risk factor based score and thus save morbidity. We have constructed a polygenic risk score (PRS) based on genetic variants associated to risk factors and outcomes of T2D available from GWAS catalog and tested it on 5,000 genotyped Caucasian subjects with T2D of ADVANCE trial and its 5 year extension in ADVANCE-ON. We selected 620 SNPs which in combination with geo-ethnic principal component, age of onset and diabetes duration achieved a highly significant prediction of outcomes, including cardiovascular (AUC=0.80) and total mortality (AUC=0.77), equalling and surpassing such clinical scores as Framingham and UKPDS. The highest positive predictive values for micro- and macro-vascular prevalent cases were 81% and 78%, respectively. Individuals within the highest tertile of genetic risk for total prevalence of both microvascular and macrovascular events, had significantly higher cardiovascular death than subjects with the lowest genetic score (p=2.02x10-12). The number needed to treat to prevent one CV death over 5 years by ADVANCE therapies, was 12 in subjects with high PRS (p=0.0045) compared to 45 (p=0.14) in subjects with low PRS. Cox cumulative hazard plots demonstrated persistence of benefits for all cause and cardiovascular mortality in high PRS tertile over 10 years of intensive blood pressure control, while for glucose intensive therapy the benefit was restricted to the high PRS group for end-stage renal disease. We conclude that the use of a PRS that detects the risk of micro- and macrovascular complications at the time of T2D diagnosis could effectively assist in preventive and therapeutic measures to attenuate diabetes complications.

Disclosure

P. Hamet: Consultant; Self; Servier. F. Harvey: None. M. Haloui: None. M. Woodward: None. R. Tahir: None. F. Marois-Blanchet: None. C. Long: None. M. Marre: Board Member; Self; Abbott. Consultant; Self; AstraZeneca. Board Member; Self; Intarcia Therapeutics, Inc.. Consultant; Self; Eli Lilly and Company. Board Member; Self; Merck Sharp & Dohme Corp., Novo Nordisk A/S. Research Support; Self; Sanofi. Board Member; Self; Servier. S. Harrap: Research Support; Self; Servier. J. Chalmers: Research Support; Self; Servier. Other Relationship; Self; Servier. J. Tremblay: Consultant; Self; Servier.
GDF15 Is Induced by T1D-Associated Cytokines and ER Stress and Promotes Beta-Cell Apoptosis
Authors: XU GUANLAN , THIELEN LANCE , CHEN JUNQIN , JO SEONGHO , SHALEV ANATH
Published: June 2018

Type 1 diabetes (T1D) is characterized by T-cell and cytokine-mediated pancreatic beta cell loss, but some of the mechanisms involved are still unknown. Growth differentiation factor 15 (GDF15), which is a distant member of the transforming growth factor β (TGFβ) family, has been shown to be increased in various diseases including cancer, heart failure and type 2 diabetes and to promote cancer cell apoptosis. Although we found that GDF15 is highly expressed in adult human beta cells, its role in beta cells and T1D has not been studied. In the present study, we found that GDF15 is significantly elevated in the islets of diabetic female NOD mice compared to nondiabetic female NOR and male NOD mice. In addition, by using human islets, Akita beta cells and INS-1 beta cells, we found that the expression and content of GDF15 are significantly increased by T1D-associated cytokines and thapsigargin/ER stress. Furthermore, we demonstrated that GDF15 knockdown significantly blunts cytokines-induced beta cell apoptosis, while GDF15 overexpression exacerbates T1D-associated cytokines-induced beta cell apoptosis. Taken together, these findings suggest that GDF15 is induced by cytokine-induced ER-stress in beta cells and might contribute to the beta cell loss in T1D. Targeting beta cell GDF15 may therefore represent a novel strategy to inhibit beta cell loss in T1D.

Disclosure

G. Xu: None. L. Thielen: None. J. Chen: None. S. Jo: None. A. Shalev: None.
Alpha-1 Antitrypsin (AAT) Protects ß Cells via Clathrin-Mediated Endocytosis
Authors: WANG JINGJING , KIM DO-SUNG , STRANGE CHARLIE , WANG HONGJUN
Published: June 2018

AAT, an acute phase reactant serine protease inhibitor, exerts protective effects on islets under stress. We reported that AAT inhibits β cell death through the suppression of cytokine-induced c-Jun N-terminal kinases (JNK) activation, in addition to its anti-inflammatory effects extracellularly in an islet transplantation model. To further understand the direct effects of AAT on β cells, we studied AAT endocytosis in human islets and a murine insulinoma cell line, βTC3 cells. We found that AAT was internalized by β cells in a time- and dose-dependent manner. AAT internalization was mediated by clathrin as treatment with chlorpromazine (CPZ), a drug that disrupts clathrin-mediated endocytosis, profoundly decreased AAT internalization. Besides, AAT inhibited cytokine-induced JNK activation, caspase 9 cleavage and DNA fragmentation, which were also overturned by the addition of CPZ. Meanwhile, although evidence suggests that clathrin interacts with autophagy related 16 like 1 (Atg16L1) which results in autophagy in several types of cells, we observed only mild induction of Atg16L1 and autophagy after AAT treatment in β cells.

In summary, as shown in the image, we found that AAT is internalized by β cells via clathrin-mediated endocytosis. Moreover, our finding that AAT endocytosis is required for its protective effects on β cells provides new insight into the usage of AAT-based therapy in diabetes and related diseases.

Disclosure

J. Wang: None. D. Kim: None. C. Strange: Research Support; Self; Grifols, CSL Behring, Shire. Consultant; Self; Shire. Stock/Shareholder; Self; Abeona. Consultant; Self; CSL Behring. Research Support; Self; Adverum. Consultant; Self; Adverum. Research Support; Self; MatRx. H. Wang: None.
Altering ß-Cell Proliferation In Vivo by Pharmacological Manipulation of the Nrf2 Pathway
Authors: KATZ LIORA S. , GARCIA-OCANA ADOLFO , SCOTT DONALD
Published: June 2018

Glucose is a natural mitogen that drives adaptive β-cell mass expansion by promoting proliferation. We previously demonstrated that induction of Nrf2 is required for glucose-stimulated and carbohydrate response element binding protein (ChREBPa)-augmented β-cell proliferation. Here we found that adenoviral delivery of Nrf2 increased human β-cell proliferation by 8.9 ± 0.2 fold (n=6), which exceeded the effect of 20 mM glucose (4.3 ± 0.4 fold). To test if activation of the Nrf2 pathway is necessary for adaptive β-cell expansion in vivo, we used pharmacological inhibitors and activators of Nrf2. Mice were placed on a high fat diet (HFD) or standard chow diet (SD) for 7 days and treated with vehicle or brusatol, which depletes Nrf2 by a post-translational mechanism. Treatment with brusatol effectively diminished Nrf2 abundance in β-cells as determined by immunofluorescence. Further, brusatol treatment ameliorated the increased β-cell proliferation normally seen with a one-week HFD (2.7±03% ki67 positive β-cells in control HFD mice, vs. 1.6±0.3% in HFD mice treated with brusatol, p < 0.05, n=5). Brusatol-treated mice also displayed significantly diminished glucose tolerance on a HFD by IPGTT, without altering the food intake or weight of the animals. Using an activator of Nrf2, bardoxolone methyl (CDDO-Me), for 7 days in the presence or absence of HFD. We found, consistent with previous reports, improved glycemic levels and decreased body weight in mice treated with CDDO-Me on a HFD. Remarkably, we also found increased β-cell proliferation in mice fed a SD and treated with CDDO-Me (1.7 ± 0.1% controls compared to 2.6±0.1% ki67 positive, insulin positive β-cells in mice treated with CDDO-Me). Crucially, in human β-cells, brusatol inhibited glucose-stimulated proliferation (1.9 fold vs. 12.7 fold, n=2) and CDDO-Me increased proliferation in 5 mM glucose (37.5 fold, n=2). Thus, we conclude that the Nrf2 pathway is an exciting novel target for manipulation of β-cell proliferation in vivo.

Disclosure

L.S. Katz: None. A. Garcia-Ocana: None. D. Scott: None.
The Patterning of Pancreatic Progenitors Is Regulated by the Temporal Control of NOTCH Signaling
Authors: BUKYS MICHAEL A. , JENSEN JAN
Published: June 2018

Pancreatic development occurs through the specification of three main lineages from a single progenitor cell. Previous studies have shown the importance of NOTCH signaling in both endocrine and exocrine differentiation during pancreatic development. Here we use the directed differentiation of pluripotent cells to demonstrate that the duration a culture remains in a NOTCH engaged progenitor state contributes to downstream lineage specification. Short-duration Pancreatic Progenitor (PP) cultures that were exposed to NOTCH inhibition showed significant up regulation of the Pro-endocrine genes NGN3, NKX2.2 and NEUROD with a corresponding activation of the endocrine genes INS, GCG and SST. The expression levels of these genes were greatly blunted when NOTCH inhibition of the PP culture was delayed. Cultures that were kept at the PP state lost endocrine competence when challenged by NOTCH inhibition, and displayed patterning towards exocrine lineages (DUCT/ACINAR) as evident through the up-regulation of CPB1, MIST1 and ONECUT1. Terminal differentiation of prolonged PP culture, adopted a ductal phenotype as evident through increased expression of F3, HNF1B and PROM1. Using the PP population in subsequent DoE based computer modeling experiments to optimize for either INS, MIST1 or F3 expression demonstrated that NKX6.1 was significantly up-regulated in short term PP cultures optimized for insulin expression as compared to the delayed cultures. Acinar conditions achieved through optimization for MIST1 expression displayed highly increased PTF1A expression. Finally, PP cultures that were maintained in the absence NOTCH inhibition and subsequently optimized for F3 expression displayed increased HNF1B and DBA expression. Altogether this suggests that the lineage specification within the pancreatic field of pluripotent derived cells is partially dictated by the duration a cell spends in a NOTCH active state, confirming findings of the embryonic murine developing organ.

Disclosure

M.A. Bukys: None. J. Jensen: Stock/Shareholder; Self; Trailhead Biosystems inc.
Noninvasive Longitudinal Evaluation of GPR-119 Agonist Effects on ß-Cell Mass Using GLP-1 Receptor-Targeting SPECT/CT
Authors: MURAKAMI TAKAAKI , FUJIMOTO HIROYUKI , FUJITA NAOTAKA , HAMAMATSU KEITA , MATSUMOTO KOJI , INAGAKI NOBUYA
Published: June 2018

Aims:Type 2 diabetes mellitus (T2DM) is known as a progressive disease with loss of pancreatic β-cell mass (BCM) and functions. However, longitudinal changes of BCM remain veiled since non-invasive techniques for observation of BCM in vivo have not been established. We developed [Lys12(111In-BnDTPA-Ahx)]exendin-4 (111In-Ex4) targeting a glucagon-like peptide 1 (GLP-1) receptor, which enabled to quantify BCM non-invasively. DS8500a, a G protein-coupled receptor 119 (GPR119) agonist, is under investigation for the treatment of T2DM. Some GPR119 agonists showed the possible stimulation of β-cell replication, which suggested some preferable effects of GPR119 agonists on BCM. Therefore, we investigated this GPR119 agonist effect on BCM in diabetic mice using our 111In-Ex4 SPECT/CT technique.

Method: Five-week-old male db/db mice with dietary restriction were assigned to the following two groups: mice fed with normal chaw (group C) and those with DS8500a-mixed chaw (group DS). We performed 111In-Ex4 SPECT/CT every four weeks until 13 weeks old and evaluated the RI accumulations in the pancreas. Ex-vivo pancreatic RI intensities, together with pathological and immunohistochemical analysis of BCM were also examined.

Results: As for 111In-Ex4 SPECT/CT, pancreatic RI intensities in group DS were significantly higher than those of group C, at the age of 9 and 13 weeks old. Moreover, ex-vivo pancreatic RI intensities in group DS were significantly higher than those of group C. Pathological BCM analysis revealed significantly larger BCM in group DS than in group C.

Conclusion: 111In-Ex4 SPECT/CT as wells as ex-vivo pancreatic analysis showed preserved BCM in mice with DS8500a. This result was corroborated by the conventional pathological method. Taken together, 111In-Ex4 SPECT/CT showed DS8500a, a GPR119 agonist, could attenuate the progression of BCM loss in diabetic mice.

Disclosure

T. Murakami: None. H. Fujimoto: None. N. Fujita: None. K. Hamamatsu: None. K. Matsumoto: None. N. Inagaki: Research Support; Self; Daiichi Sankyo Company, Limited, AstraZeneca, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Mitsubishi Tanabe Pharma Corporation, Takeda Pharmaceutical Company Limited, Japan Tobacco Inc., MSD K.K., Sumitomo Dainippon Pharma Co., Ltd., Nippon Boehringer Ingelheim Co. Ltd., Novartis Pharma K.K., Kissei Pharmaceutical Co., Ltd., Novo Nordisk Pharma Ltd., Eli Lilly and Company, Sanwa Chemical Industrial Co., Ltd., Teijin Pharma Limited.
Unusual Duct-Like Cells in the Islet of Diabetic C414A-CRY1 Transgenic Mice
Authors: OKANO SATOSHI , YASUI AKIRA , KANNO SHIN-ICHIRO , SATOH KENNICHI , IGARASHI MASAHIKO , NAKAJIMA OSAMU
Published: June 2018

Cryptochrome (CRY) proteins play indispensable roles in the mammalian circadian clock. We previously generated transgenic mice ubiquitously expressing mCRY1 with a mutation in cysteine414 (the zinc-binding site of CRY1). The Tg mice overexpressing the mutant CRY1 (C414A-CRY1) showed early onset diabetes mellitus characterized by β-cell dysfunction in addition to unusual circadian rhythms in locomotor activities. We have already shown that the decrease of insulin secretion from β-cells along with the lowered proliferation of β-cells due to the senescence-associated secretory phenotype (SASP)-like changes in the Tg mice (Okano S., 2016). We have also demonstrated that, atypical duct-like structures having some common morphological characters to pancreatic intraepithelial neoplasias (PanINs) that can cause pancreatic ductal adenocarcinoma, frequently appeared in the aged Tg mice. In this study, to explore the relation between the unique features of β-cells and the generation of PanIN-like structures, we further performed detailed studies of the pancreases of Tg mice. We found that unusual duct-like structures producing mucin emerged not only in the exocrine areas but also in the inside islets in the Tg mice frequently with age. These results may suggest that islet cells in the Tg mice can transdifferentiate to duct-like cells, thereby contributing to generate PanIN-like lesions in the Tg mice. In addition, the extent of fibrosis in the islet increased in the Tg mice compared with wild type controls. Taken together, our results suggest the SASP-like microenvironment in the islet play some roles to modify the integrity of endocrine cells as well as the architecture of the islet.

Disclosure

S. Okano: None. A. Yasui: None. S. Kanno: None. K. Satoh: None. M. Igarashi: None. O. Nakajima: None.
T3 and Glucose Increase Expression of ChREBPß to Induce Expression of Phosphoenolpyruvate Carboxykinase (Pck1) Leading to Increased ß-Cell Proliferation
Authors: KATZ LIORA S. , SCOTT DONALD
Published: June 2018

Thyroid hormone (T3) is required for islet development and function. Both T3 and high glucose concentrations are critical components of differentiation protocols of stem cells to β-cells. We previously demonstrated that in brown adipose tissue, T3 and glucose synergistically regulate carbohydrate response element binding protein (ChREBP), which in turn upregulates Ucp1, Glut4 and Fasn, resulting in increased thermogenesis, decreased body weight and improved glycemic levels. In the present study, we asked whether T3 and glucose signaling pathways can coordinately regulate transcription of genes important for β-cell function and mass. RNA-seq analysis was performed on cadaveric human islets from five different donors in response to low and high glucose concentrations (6 and 20 mM, respectively) and in the presence or absence of T3 (10 nM). We found that T3 and glucose coordinately regulate the expression of ChREBPβ and Pck1 (phosphoenolpyruvate carboxykinase-1). Typically, it is thought that Pck1 is not expressed in pancreatic β-cells, but it is a known target of T3 in hepatocytes. However, in FACS sorted adult human β-cells, we found that high concentrations of glucose (20 mM) and T3 (10 nM) induced the expression of Pck1 by 17.2±1.3 fold for mRNA levels. In addition, we found that overexpression of Pck1 together with its substrate dimethyl malate (DMM) increased β- cell proliferation in human islets (0.49±0.03% for human islets overexpressing Pck1, treated with DMM compared with 0.01±0.01% for control islets, n=4). Finally, using a sh-RNA knockdown approach, we demonstrated that in Ins-1 cells, ChREBPβ is necessary for Pck1 dependent β-cell proliferation. We conclude that T3 and glucose act together to regulate ChREBPβ which in turn upregulates expression of Pck1. In turn, Pck1 activity leads to increased β-cell mass.

Disclosure

L.S. Katz: None. D. Scott: None.
A Novel Mediator of Arsenic-Induced Pancreatic Beta-Cell Dysfunction
Authors: CARMEAN CHRISTOPHER M. , YOKOI NORIHIDE , KIRKLEY ANDREW G. , TAKAHASHI HARUMI , SARGIS ROBERT M. , SEINO SUSUMU
Published: June 2018

With up to 140 million people living above arsenic (AS)-contaminated groundwater, AS exposure presents a major health risk worldwide. Studies have revealed a positive correlation between AS exposure and diabetes, where those suffering from symptoms of chronic AS toxicity have 5-6 times the local rates of diabetes. Recent mouse models of AS exposure suggested that this form of diabetes may be mediated by pancreatic β-cell dysfunction. To evaluate this possibility, the MIN6-K8 mouse β-cell line was utilized as a model. Three days of physiologically-relevant AS exposure decreased glucose-induced insulin secretion (GIIS) without effects on basal secretion. Three days of exposure to 1 µM AS decreased GIIS more than 60% without effects on cell count, total protein, total DNA, mitochondrial mass, or O2 consumption. RNAseq analysis of AS-exposed MIN6-K8 cells revealed 3 significantly regulated genes previously unassociated with β-cell function.

Each gene was individually knocked down by siRNA in the presence of AS. Knockdown of one of these genes, UDP-glucuronosyltransferase 1a6a (Ugt1a6a), partially recovered GIIS in AS-exposed cells. Ugt1a6a is a phase 2 detoxification enzyme for the conjugation of small aromatic compounds. The precursor and rate-limiting factor for serotonin production, 5-hydroxytryptophan (5-HTP), a known target of Ugt1a6a and modulator of GIIS, was decreased following AS exposure. Therefore, the AS-mediated increase in Ugt1a6a expression may deplete serotonin, and secondarily 5-HTP, decreasing GIIS. This work introduces a gene with a novel role in β-cell physiology and arsenic-mediated pathology.

Disclosure

C.M. Carmean: None. N. Yokoi: None. A.G. Kirkley: Consultant; Self; AbbVie Inc., Takeda Pharmaceuticals U.S.A., Inc.. H. Takahashi: None. R.M. Sargis: Advisory Panel; Self; CVS/Caremark. S. Seino: Research Support; Self; Merck Sharp & Dohme Corp., MSD K.K., Novo Nordisk Inc., Poxel SA, Sumitomo Dainippon Pharma Co., Ltd., Sanofi K.K.. Speaker's Bureau; Self; Sumitomo Dainippon Pharma Co., Ltd.. Research Support; Self; Daiichi Sankyo Company, Limited. Speaker's Bureau; Self; Novartis Pharma K.K.. Research Support; Self; Taisho Pharmaceutical Co., Ltd..
Analysis of ∼700 Human Tissue Samples Identifies microRNAs That Are Associated with, Predictive of, and Necessary for Insulin Gene Transcription
Authors: HARDIKAR ANAND , WONG WILSON , JOGLEKAR MUGDHA , DALGAARD LOUISE T. , JENKINS ALICIA , MA RONALD C. ,   THE MICRORNA ATLAS STUDY GROUP
Published: June 2018

There is growing evidence that microRNAs, a class of non-coding RNA molecules, are associated with regulation of gene expression in general. Here we profiled microRNAs and key endocrine pancreatic gene transcripts in 698 human tissues, including ∼200 human islet preparations. Figure 1 shows the tissue types in our biobank and circos plots showing correlation of key pancreatic genes and microRNAs. Using novel approaches involving penalized regression and bootstrapping in our discovery set of 507 tissue samples, we identified microRNAs that are highly associated with (pro-)insulin expression. Then, using two different de-identified validation sets (N=91 and N=100) of human cells/tissue samples, we demonstrate that the levels of insulin-associated microRNAs can predict the level of insulin transcript with high accuracy. Finally, we show that forced expression of insulin gene does not change the levels of insulin-associated microRNAs, but forced expression of selected microRNAs promotes insulin gene expression. More generally, these data identify key microRNA genes associated with, predictive of and promoting for insulin gene transcription.

Disclosure

A. Hardikar: None. W. Wong: None. M. Joglekar: None. L.T. Dalgaard: Employee; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. A. Jenkins: Research Support; Self; Medtronic, Mylan, Sanofi-Aventis. R.C. Ma: Research Support; Self; AstraZeneca, Bayer AG, Merck Sharp & Dohme Corp., Pfizer Inc.. Advisory Panel; Self; Boehringer Ingelheim GmbH, Nippon Boehringer Ingelheim Co. Ltd..
Newly Revealed Role for OGT in Obesity-Precipitated ß-Cell Hyperinsulinemia
Authors: LOCKRIDGE AMBER D. , JO SEOKWON , DAMBERG NIKLAS E. , MOHAN RAMKUMAR , ALEJANDRO EMILYN
Published: June 2018

Highly expressed in the β-cell, the nutrient sensor O-GlcNAc transferase (OGT) couples metabolic environment to cellular behavior through dynamic protein modifications (O-GlcNAcylation) but the role of OGT in β-cell hyperlipidemia is unknown. Here, we observed changes in islet O-GlcNAcylation in obese humans and high-fat diet (HFD, 60%) fed mice vs. lean controls, leading us to hypothesize that OGT mediates β-cell adaptations to diet-induced obesity. In a mouse model of partial β-cell OGT loss, we found that Rip-Cre; OGT f/+ (OGT Het) mice failed to develop hyperinsulinemia over 12 weeks of HFD. Only Het islets isolated from HFD mice showed deficits in stimulated insulin secretion (30 mM KCl, vs. HFD Ogt f/+). Nutrient-driven ATP was not impaired, but islets with full β-cell OGT KO (Rip-Cre; OGT f/f) showed no acute secretory potentiation to palmitate (100 μM) implicating the amplification arm of the stimulus-secretion pathway. HFD β-cell mass was similar (OGT Het vs. WT) but increased Ins+/Gcg+ cells and Ngn3 gene expression in OGT Het islets suggested functional β-cell loss through enhanced dedifferentiation. HFD hypoinsulinemia was reiterated in a tamoxifen-induced OGT loss model (Mip-Cre; OGT f/f), which also showed elevated proinsulin/insulin serum ratios (6 weeks HFD). Transcriptome analysis by RNA-sequencing of OGT KO and WT mouse islets revealed differential gene expression of proteins involved in membrane depolarization (Kcnj11, Abcc8), ER calcium (Atp2a2), lipid uptake (Lpl) and β-cell differentiation (Ucn3). An upstream analysis suggested impairments in free radical scavenging, supported by elevated levels of 4-HNE lipid peroxidation staining in post-HFD OGT Het islets. However, antioxidant supplementation of HFD chow (0.2% alpha-lipoic acid) did not rescue OGT Het in vivo deficits. These data reveal OGT as a critical mediator of β-cell adaptation to diet-induced obesity and highlight protein O-GlcNAcylation as a potential therapeutic target to prevent prediabetic progression.

Disclosure

A.D. Lockridge: None. S. Jo: None. N.E. Damberg: None. R. Mohan: None. E. Alejandro: None.