bluuebunny/crossref_metadata_2025_split huggingface dataset:
237-OR: Insulin Icodec: An Insulin Analog Suited for Once-Weekly Dosing in Type 2 Diabetes
Authors: HÖVELMANN ULRIKE , BRØNDSTED LISE , KRISTENSEN NIELS R. , RIBEL-MADSEN RASMUS , DEVRIES J. HANS , HEISE TIM , HAAHR HANNE 
Published: June 2020

Insulin icodec* is a novel basal insulin analog designed for single once-weekly (OW) subcutaneous injection. This randomized, double–blind, double–dummy trial investigated the pharmacokinetics (PK), pharmacodynamics (PD) and safety of insulin icodec. Fifty individuals with type 2 diabetes (insulin treated±metformin; 43 men; mean±SD age 57±5 years, BMI 30.1±2.7 kg/m2, A1C 7.4±0.6%) received OW insulin icodec (12, 20 or 24 nmol/kg) plus once–daily (OD) placebo (N=13, 13, 12) or OD insulin degludec (0.4 U/kg) plus OW placebo (N=12) for 5 weeks in 3 dose level cohorts. PD properties were investigated at close to steady state in 24-h glucose clamps on days 2 and 7 after the last insulin icodec dose. Median tmax,insulin icodec was 16 h and geometric mean t½,insulin icodec was 196 h with no systematic differences between dose levels. The glucose–lowering effect over a weekly dosing interval was derived from the observed PK/PD data using a PK/PD model and showed close to even distribution over the 7 days across all dose levels (Figure). The adverse event (AE) incidence did not increase with increasing insulin icodec dose (100%, 69%, 75%, respectively). There were no serious or severe AEs, severe hypoglycemic episodes or injection site reactions.
In conclusion, insulin icodec was safe and well-tolerated and showed PK/PD properties supporting once-weekly administration at clinically relevant dose levels. * Proposed INN.

Disclosure
U. Hövelmann: None. L. Brøndsted: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. N.R. Kristensen: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. R. Ribel-Madsen: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. J. DeVries: Advisory Panel; Self; Novo Nordisk A/S, Zealand Pharma A/S. Consultant; Self; Metronom Health. Employee; Self; Profil Institute for Clinical Research. T. Heise: Advisory Panel; Self; Mylan, Novo Nordisk A/S. Research Support; Self; ADOCIA, Aerami, Becton, Dickinson and Company, Biocon, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Gan & Lee Pharmaceuticals, MedImmune, Merck KGaA, Mylan, Nordic Bioscience, Novo Nordisk A/S, Poxel SA, Sanofi, Xeris Pharmaceuticals, Inc., Zealand Pharma A/S. Speaker’s Bureau; Self; Eli Lilly and Company, Novo Nordisk A/S. H. Haahr: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S.

Funding
Novo Nordisk
238-LB: Dietary Protein Restriction Induces Myocardial Dysfunction Despite Reducing Body Weight in Aged C57BL/6J Mice
Authors: AXELROD CHRISTOPHER L. , DANTAS WAGNER S. , DAVULURI GANGARAO , KING WILLIAM T. , HILL CRISTAL M. , MORRISON CHRISTOPHER , KIRWAN JOHN P. 
Published: June 2020

Introduction: Type 2 diabetes (T2D) and obesity reduce healthspan in part by increasing risk of cardiovascular events including arrhythmias, heart attacks, cardiomyopathies, and coronary heart disease. Low protein diets and specific amino acid deficiencies, such as dietary methionine, have been observed to reduce body weight, improve glycemic control, and ultimately extend lifespan. However, little is known of how systemic protein deficiency may alter myocardial integrity and bioenergetic function.
Methods: Twelve-month-old male C57BL/6J mice were randomized to a normal protein (NP; 20% casein), low protein (LP; 5% casein), high fat (HF; 60% fat), or high fat + low protein (HF + LP; 60% fat, 5% casein) diet for 16 weeks. Body weight was measured weekly. Oxidative phosphorylation (OXPHOS) and electron transport system (ETS) capacity was determined in permeabilized myocardial fiber bundles by high-resolution respirometry. Citrate synthase activity was determined by kinetic enzymatic assay. Cardiomyocyte diameter was determined by haemotoxylin and eosin (H&E) staining. Untargeted transcriptomic profiling was conducted by RNA sequencing. Changes in protein expression were determined by immunoblotting.
Results: LP reduced body weight relative to NP and HF, whereas HF+LP reduced body weight relative to HF, but was greater than LP alone. LP and LP+HF increased heart weight relative to body weight and cardiomyocyte diameter relative to both NP and HF. Succinate linked OXPHOS and ETS (complex II coupled and uncoupled) was reduced in both LP and LP+HF compared to NP and HF alone.
Conclusion: Protein restriction reduces cardiac OXPHOS capacity and induces cardiomyocyte hypertrophy in aging mice. These results question the applicability of chronic protein restriction for management of obesity-related diseases. Further investigation is required to determine if the alterations in cardiac bioenergetic function and structure are advantageous or maladaptive responses.

Disclosure
C.L. Axelrod: None. W.S. Dantas: None. G. Davuluri: None. W.T. King: None. C.M. Hill: None. C. Morrison: None. J.P. Kirwan: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (DK105032); National Institute of General Medical Sciences (GM104940)
240-LB: Plant-Based Flours Reduce Body Weight and Adiposity in High-Fat Fed Rats
Authors: MARTINEZ TAYLOR , DUCA FRANK , WENINGER SAVANNA N. , MEYER RACHEL , LANE ADELINA I.L. 
Published: June 2020

The relationship between obesity and the gut microbiome has become increasingly recognized, highlighting the need to determine how specific diets can beneficially impact gut microbiota to improve metabolic disease. Many studies have found an increase in dietary fiber not only causes a shift in the microbiome but also results in weight loss and altered feeding behavior. Dietary fibers are diverse both in composition and source, being found in many different plant-based flours. Fiber can also be characterized by solubility, with highly soluble fibers being most easily broken down by gut bacteria to produce short chain fatty acids (SCFAs). However, the efficacy and mechanism of specific plant-based flours on obesity is not well characterized. To better understand which plant-based flours have the greatest impact on energy homeostasis, we treated high-fat diet (HFD; 45% kcal from diet)-fed rats with four different flours for 6wks. The flours, barley flour, oat bran, wheat bran, and amylose maize were incorporated into the HFD at either 5% or 10% fiber content, and were compared to an isocaloric, nutrient-matched chow and HFD controls. We hypothesized that the soluble barley flour at 10% fiber would have the greatest impact on energy homeostasis due to the high levels of beta-glucan, while the amylose maize, oat bran, and wheat bran will have a significant but less pronounced effect. We found that the rats fed high dose barley flour and wheat bran had significantly reduced body weight gain. High and low dose barley flour resulted in significantly reduced food intake compared to HFD, and all high dose flour diet-fed rats exhibited lower adiposity compared to the HFD rats. We found all high dose flour diet-fed rats exhibited changes in feeding behavior and energy expenditure compared to HFD rats measured by metabolic cages. These results demonstrate plant based fibers can have differing effects on food intake and body composition, underscoring the need to determine the most effective dietary manipulations for obesity treatment.

Disclosure
T. Martinez: None. F. Duca: None. S.N. Weninger: None. R. Meyer: None. A.I.L. Lane: None.

Funding
National Institute of Food and Agriculture (2019-67017-29252)
243-OR: Integrative Multiomics Analysis of Glucometabolic Traits, Adipose Tissue DNA Methylation, and Gene Expression Identifies Epigenetic Regulatory Mechanisms of Insulin Resistance and Obesity in African Americans
Authors: SHARMA NEERAJ K. , COMEAU MARY , MONTOYA DENNIS J. , PELLEGRINI MATTEO , HOWARD TIMOTHY , LANGEFELD CARL D. , DAS SWAPAN K. 
Published: June 2020

Decline in insulin sensitivity due to dysfunction of adipose tissue (AT) is one of the earliest pathogenic events in type 2 diabetes. We hypothesize that differential DNA methylation (DNAm) in regulatory regions of a subset of genes controls insulin sensitivity and obesity by modulating transcript expression in AT. Integrating reduced representation bisulfite sequencing-derived AT DNAm profiles for 1,054,719 CpG sites and 200,800 CpG regions with AT transcript profile data in 230 African Americans, we performed cis-expression quantitative trait methylation (cis-eQTM) analysis focused on 6,774 glucometabolic trait-associated transcripts to identify local epigenetic regulatory loci for these transcripts. We identified significantly associated CpG regions for 140 transcripts (FDR-P&lt;0.1). The strongest eQTM locus was observed for the proopiomelanocortin (POMC; ρ= -0.632, P = 4.70X10-27) gene at the chr2:25,384,001-25,385,000 region covering a CpG island in the gene. Epigenome-wide association studies (EWAS) further examined the association of DNAm levels of CpG sites/regions with insulin sensitivity (Matsuda index and SI) and BMI. EWAS identified 520, 154, and 482 CpG regions associated (FDR-P &lt;0.1) with Matsuda index, SI and BMI, respectively. We jointly analyzed the EWAS, transcript level to trait association, and eQTM data and identified significant eQTM loci for 26, 15 and 25 genes that were also associated with Matsuda index, SI and BMI, respectively in EWAS. These associated genes include fructose-2,6-biphosphatase 3 (PFKFB3), integrin alpha M (ITGAM), integrin alpha X (ITGAX), and POMC.
In summary, applying an integrative multi-omics approach, our study provides evidence for DNAm-mediated regulation of gene expression at both previously identified and novel loci for many key AT transcripts influencing insulin resistance and obesity.

Disclosure
N.K. Sharma: None. M. Comeau: None. D.J. Montoya: None. M. Pellegrini: None. T. Howard: None. C.D. Langefeld: None. S.K. Das: None.

Funding
American Diabetes Association (1-18-ICTS-113 to S.K.D.); National Institutes of Health (R01DK090111)
246-OR: A Loss-of-Function Mutation in the Sucrase-Isomaltase Gene Is Linked to a Markedly Healthier Metabolic Profile in Greenlanders
Authors: JØRGENSEN MARIT E. , ANDERSEN METTE K. , SKOTTE LINE , JØRSBOE EMIL , GRARUP NIELS , BJERREGAARD PETER , SOBORG BOLETTE , PEDERSEN OLUF , FEENSTRA BJARKE , FÆRGEMAN NILS J.K. , KOCH ANDERS , MOLTKE IDA , HANSEN TORBEN , ALBRECHTSEN ANDERS 
Published: June 2020

Objective: Congenital sucrase-isomaltase deficiency is prevalent in Arctic populations. Recently, a frameshift mutation, c.273-274delAG, in the sucrose-isomaltase encoding gene (SI) was identified. This mutation was predicted to cause complete loss of enzymatic function, and thus is a diagnostic marker for Congenital sucrase-isomaltase deficiency. We aimed to characterize the metabolic health among adult Greenlanders carrying the c.273-274delAG mutation.
Material and Methods: We genotyped the c.273-274delAG mutation in two cohorts of Greenlanders comprising 4600 (Cohort I) and 1500 participants (Cohort II), respectively. We assessed the effect of the variant on markers of metabolic health.
Results: In cohort I, homozygous carriers of the c.273-274delAG mutation had a markedly healthier metabolic profile than the remaining population, including lower BMI (beta, -2.0 kg/m2, p=3.1x10-5), fat% (-3.3 %, p=0.0004), weight (-4.8 kg, p=0.0005), and levels of triglycerides (-0.27 mmol/l, p=2.3x10-6) and remnant cholesterol (-0.11 mmol/l, p=4.2x10-5). Even though homozygous carriers had a significantly lower consumption of sugar, this did not fully explain the healthier phenotype in these individuals. In cohort II, metabolomics data revealed that particularly HDL metabolism was altered among homozygous carriers of the mutation, and that these individuals had markedly increased levels of circulating acetate (1.78 mmol/l, p=2.1x10-26). Based on these findings, we hypothesize that the observed phenotype is caused by increased fermentation of undigested carbohydrates in the gut, leading to increased amounts of circulating acetate, which increases lipid oxidation and reduces storage of fat.
Conclusion: Our results show, that homozygous c.273-274delAG mutation carriers have a markedly healthier metabolic profile, and indicate that SI constitutes a promising new drug target, with potential to prevent obesity and dyslipidemia.

Disclosure
M.E. Jørgensen: Research Support; Self; Amgen, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi-Aventis. Stock/Shareholder; Self; Novo Nordisk A/S. M.K. Andersen: None. L. Skotte: None. E. Jørsboe: None. N. Grarup: None. P. Bjerregaard: None. B. Soborg: None. O. Pedersen: None. B. Feenstra: Employee; Spouse/Partner; Novo Nordisk Inc. N.J.K. Færgeman: None. A. Koch: None. I. Moltke: None. T. Hansen: None. A. Albrechtsen: None.
247-OR: ADA Presidents’ Select Abstract: Detection of T-Cells Reactive to Hybrid Insulin Peptides in Subjects at Risk for Type 1 Diabetes
Authors: BAKER ROCKY L. , DELONG THOMAS , REWERS MARIAN , GOTTLIEB PETER , HASKINS KATHRYN M. 
Published: June 2020

Our lab recently established that Hybrid Insulin Peptides (HIPs) are antigenic ligands for diabetogenic T cell clones in the NOD mouse model of autoimmune diabetes. Importantly, we have found that the phenotype of HIP-reactive T cells present in the peripheral blood of NOD mice is indicative of ongoing autoimmunity in the pancreas. In the human setting, we demonstrated that T cells reactive to HIPs are present in the residual islets of type 1 diabetic (T1D) organ donors and more recently, in PBMC from T1D patients. Using an IFN-gamma ELISPOT assay to test T cell reactivity in the peripheral blood of T1D patients or control subjects to a panel of 16 HIPs, we demonstrated that 70% of the responding patients showed reactivity to multiple HIPs. To determine whether the presence of HIP-reactive T cells might serve as a biomarker of disease activity in humans, we are following longitudinally a cohort of subjects positive for multiple islet autoantibodies (Ab+), individuals that are at high risk for developing T1D. Using IFN-gamma ELISPOT and CFSE dilution assays, our data indicate that HIP-reactive T cells with an inflammatory phenotype can be detected before disease onset. In one Ab+ individual that showed HIP reactivity, we used single-cell RNA sequencing to study the phenotype and TCR usage of HIP-responding T cells in 3 longitudinal samples collected 6 months apart. We observed that T cells with a regulatory phenotype (FoxP3+, CTLA-4+, LAG3+) can be observed when a proliferative response - but not inflammatory - is detected. In this ongoing study, we are reporting for the first time that HIP-reactive T cells can be observed in the peripheral blood of subjects at risk for T1D, and our data may indicate that the balance of inflammatory vs. regulatory signals of HIP-reactive T cells could be potential indicators of disease activity.

Disclosure
R.L. Baker: None. T. Delong: None. M. Rewers: None. P. Gottlieb: Advisory Panel; Self; Janssen Scientific Affairs, LLC., Tolerion, Inc., Viacyte, Inc. Research Support; Self; Caladrius Biosciences, Inc., Immune Tolerance Network, Janssen Pharmaceuticals, Inc., JDRF, Leona M. and Harry B. Helmsley Charitable Trust, Mercia/Nova Laboratories, National Institute of Diabetes and Digestive and Kidney Diseases, Novo Nordisk Inc. Stock/Shareholder; Self; IM Therapeutics. K.M. Haskins: None.

Funding
American Diabetes Association (1-15-ACE-14 to T.D.); JDRF (2-SRA-2017-511-S-B); National Institutes of Health (R21AI133059-01A1)
251-LB: Changes in Alpha-Cell Function in Defective Glucose Counterregulation
Authors: PANZER JULIA , TAMAYO ALEJANDRO M. , CAICEDO ALEJANDRO 
Published: June 2020

Increased glucagon secretion from the alpha cell is the first and most important defense against hypoglycemia. In type 1 diabetes this defensive mechanism is lost, increasing the mortality risk. Understanding what goes awry with alpha cells during disease progression is therefore of utmost importance. However, studying alpha cell physiology in type 1 diabetes has met major technological roadblocks, as methods conventionally used are not appropriate or are very difficult to apply to type 1 diabetic donors. Our project overcomes these limitations by using living pancreas slices, which allows functional assessments of damaged and infiltrated islets within their native environment. By destroying beta cells with streptozotocin (STZ) we established a mouse model in which counterregulatory responses to hypoglycemia are defective 2 weeks after treatment, as shown by insulin tolerance tests in vivo. To monitor alpha cell function, we prepared pancreas tissue slices from mice expressing a genetically encoded Ca2+ indicator in all alpha cells (GCaMP6). We measured Ca2+ responses in alpha cells in response to changes in glucose concentration, glutamate receptor agonists, GABA, serotonin, somatostatin, and KCl depolarization. When comparing alpha cell responses in slices obtained from STZ-treated mice to those in untreated mice, we found that fewer alpha cells responded. Responses to glutamate receptor agonists were particularly diminished, suggesting that autocrine alpha cell activation by glutamate is disrupted. We are currently testing human slices from control and type 1 diabetic donors to determine changes in alpha cell function.

Disclosure
J. Panzer: None. A.M. Tamayo: None. A. Caicedo: None.

Funding
National Institutes of Health (R33ES025673, U01DK120456); The Leona M. and Harry B. Helmsley Charitable Trust (R-1912-03552)
252-OR: Failed Genetic Protection: Type 1 Diabetes in the Presence of HLA-DQB1*06:02
Authors: SIMMONS KIMBER , MITCHELL ANGELA M. , ALKANANI AIMON , MCDANIEL KRISTEN , BASCHAL ERIN , ARMSTRONG TAYLOR , PYLE LAURA , YU LIPING , MICHELS AARON W. 
Published: June 2020

Human leucocyte antigen (HLA) class II genes confer significant genetic risk for type 1 diabetes (T1D) development and protection. The dominant protection afforded by the DRB1*15:01 (DR15)-DQA1*01:02-DQB1*06:02 (DQ6) haplotype across all stages of T1D is well documented. This is a common haplotype found in Caucasians but present in &lt;1% of people with T1D. Knowing which metabolic, immunologic and genetic features are unique to individuals who fail genetic protection and develop T1D is important for defining the underlying mechanisms of DQB1*06:02-mediated protection. We describe a T1D cohort with DQB1*06:02 (n=50) and compare them to a T1D cohort lacking DQB1*06:02 (n=2,759), which were both identified over the last 26 years at the Barbara Davis Center, a large tertiary referral center in the Rocky Mountain Region. As expected, the average A1C was 10.8 ± 2.8% at diagnosis in 0602+ individuals. The age at diagnosis was similar between the 0602+ and 0602- participants (10.8 ± 6.0 vs. 12.5 ± 8.7 years, respectively, p=0.069) and normally distributed throughout childhood and early adulthood. The majority of 0602+ individuals with T1D were positive for one or more islet autoantibodies; however, there was a greater proportion that were islet autoantibody negative compared to 0602- individuals with T1D (p=0.004). The presence of autoantibodies directed against insulinoma antigen-2 (IA-2) was less common in 0602+ individuals (p=0.005), while the percentage of antibodies directed against GAD65, insulin and zinc transporter 8 were similar between 0602+ and 0602- individuals. Interestingly, the DQ allele on the other chromosome, DQB1*03:02 (DQ8) which confers significant T1D risk, was present in 0602+ individuals with islet autoantibodies and not in individuals without autoantibodies (p=0.002).
In conclusion, studying the clinical, immunologic and genetic features of individuals that failed genetic protection holds promise for understanding mechanisms that ultimately lead to beta cell destruction in T1D.

Disclosure
K. Simmons: None. A.M. Mitchell: None. A. Alkanani: None. K. McDaniel: None. E. Baschal: None. T. Armstrong: None. L. Pyle: None. L. Yu: None. A.W. Michels: Other Relationship; Self; ImmunoMolecular Therapeutics, LLC.

Funding
National Institutes of Health (DK095995, DK108868, DK110845, DK032083, K12DK094712-08); JDRF/Children’s Diabetes Foundation (2-SRA-2016-202-5-B); Colorado Clinical and Translational Science Institute (TR001082)
253-OR: Circulating Neutrophil Numbers Increase with Age, but Are Not a Biomarker of T1D Risk
Authors: LIN ANDREA , PERRY DANIEL J. , SCHATZ DESMOND , BRUSKO TODD M. , ATKINSON MARK A. , HALLER MICHAEL J. , MATHEWS CLAYTON E. 
Published: June 2020

Investigations into the role of the neutrophil in type 1 diabetes (T1D) have provided controversial results. It is thus essential to understand how neutrophils change with age and if these numbers are linked by T1D. To do this, we utilized whole blood count data from our database, which includes 2619 unique individuals, pancreata slices from nPOD donors, and data points from the Neulasta in T1D clinical trial, NCT00662519. We first examined changes in neutrophils, total white blood cells (WBC), and lymphocytes in individuals with T1D, T2D, controls, and first and second-degree relatives. One way ANOVA showed no significant differences in neutrophils or other WBCs between groups (p&gt;0.05). To account for age-specific effects, we built a quantile regression model using only the control subjects. With this model, a significant age association for neutrophil count was detected and repeated in each clinical group, indicating that age-matching is essential. Comparisons across groups showed that age-normalized neutrophil counts were no different between individuals with T1D, at-risk for T1D, and controls (p&gt;0.05, Mann-Whitney test). We also enumerated neutrophil numbers within the endocrine and exocrine regions of the pancreas. We assessed pancreata from 51 organ donors with and without T1D: there were no observed differences between the groups (p&gt;0.05, one-way ANOVA). Finally, data from the Neulasta trial demonstrated an increase in neutrophil numbers, but no change in the rate of C-peptide loss. We interrogated granulocytic, T cell, and dendritic cell profiles post-Neulasta to determine if changes in these subsets were reflective of treatment effects. While there was an increase in neutrophils immediately after Neulasta, changes did not persist outside of the treatment window. Other blood cell types were not responsive to Neulasta. Together, these data show that neutrophil numbers increase with age and when age correction is applied, the neutrophil is not a reliable biomarker of T1D.

Disclosure
A. Lin: None. D.J. Perry: None. D. Schatz: None. T.M. Brusko: Board Member; Self; OneVax, LLC. Consultant; Self; Caladrius Biosciences, Inc., Sanofi-Aventis. Speaker’s Bureau; Self; CSL Behring. M.A. Atkinson: None. M.J. Haller: Advisory Panel; Self; SAB Biotherapeutics, Inc. C.E. Mathews: None.