bluuebunny/crossref_metadata_2025 huggingface dataset:
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<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 <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 <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>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>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>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.
254-LB: Characterization of Ghrelin Receptor Expression in Mouse Islets Reveals Pancreatic Polypeptide Cells as a Key Ghrelin Target
Authors: GUPTA DEEPALI , MANI BHARATH K. , SHANKAR KRIPA , OSBORNE-LAWRENCE SHERRI , METZGER NATHAN , ZIGMAN JEFFREY M.
Published: June 2020

Objective: While expression of the ghrelin receptor, GHSR (growth hormone secretagogue receptor), by pancreatic islets has been investigated, data regarding the specific islet cell types that express GHSR are inconsistent and incomplete. For instance, transcriptomic studies indicate marked GHSR expression by somatostatin (SST)-secreting delta cells vs. little to no GHSR expression by alpha cells and beta cells, which contrasts studies suggesting direct actions of ghrelin on GHSRs expressed by alpha cells and beta cells. Also, to our knowledge, GHSR expression by pancreatic polypeptide (PP) cells has not previously been directly investigated. The aims of this study were to determine GHSR expression within the four principal islet endocrine cell types and the effects of ghrelin on circulating PP.

Methods: Ghsr-IRES-Cre mice, which express Cre recombinase directed by the GHSR promoter, crossed to Cre-dependent ROSA26-YFP reporter mice were used to histologically identify GHSR expression in islets. Double and triple labeling for YFP-immunoreactivity (IR) with insulin-IR, glucagon-IR, SST-IR, and/or PP-IR was performed to determine GHSR co-expression with islet hormones. Pharmacological manipulations were used to examine in vivo effects of acyl-ghrelin on plasma PP.

Results: Eighty-five % of YFP-IR cells expressed PP-IR, 50% expressed SST-IR, and none expressed insulin-IR or glucagon-IR. GHSR antagonist raised plasma PP in overnight-fasted C57BL/6N mice. However, acyl-ghrelin administration had no effect on plasma PP in C57BL/6N mice.

Conclusions: These results suggest that within mouse pancreatic islets, GHSR expression occurs most abundantly in PP cells and that pharmacologic blockade of ghrelin action raises plasma PP.

Disclosure

D. Gupta: None. B.K. Mani: None. K. Shankar: None. S. Osborne-Lawrence: None. N. Metzger: None. J.M. Zigman: Stock/Shareholder; Self; Medtronic.

Funding

Diana and Richard C. Strauss Professorship in Biomedical Research; Mr. and Mrs. Bruce G. Brookshire Professorship in Medicine; Kent and Jodi Foster Distinguished Chair in Endocrinology; University of Texas Southwestern Medical Center (to J.M.Z.)
256-OR: Using Human Induced Pluripotent Stem Cell Derived Organoids to Identify New Pathologies in Patients with Pdx1 Mutations
Authors: KRISHNAMURTHY MANSA , BRODA TAYLOR R. , KECHELE DANIEL O. , ZHANG XINGHAO , PALERMO JOSEPH J. , COLLINS MARGARET H. , THOMAS INAS H. , HEIDER AMER , DAUBER ANDREW , WELLS JAMES
Published: June 2020

Pancreas and gastrointestinal (GI) tract development is guided by several transcription factors, including pancreatic and duodenal homeobox gene-1 (PDX1). While it is well known that PDX1is essential for pancreatic development and beta cell function, its role in GI development and function is not fully understood. We identified a patient with a homozygous point mutation in PDX1 causing total loss of PDX1 protein resulting in pancreatic agenesis. We differentiated induced pluripotent stem cells from the patient into human antral gastric (HAGO) and intestinal (HIO) organoids as a novel diagnostic approach to identify new GI pathologies. Immunofluorescence staining revealed that portions of the PDX1-null HAGOs lost the gastric marker claudin18 and increased intestinal villin1 expression, signifying that regions of the antrum are converting to an intestinal phenotype. Similarly, PDX1-null HIOs lost the intestinal marker cadherin17 and increased claudin18, indicating that regions of the intestine are undergoing gastric metaplasia. Examination of functional markers revealed an increase in acid producing parietal cells, complete lack of gastrin producing endocrine cells in HAGOs, and an increase in gastric surface mucin in HIOs. These in vitro results were further validated using endoscopy biopsies from two patients with PDX1 mutations, which showed a similar disruption in gastric and intestinal identity. CRISPR/Cas9-mediated correction of the PDX1 mutation reversed the observed PDX1-null metaplasic phenotypes in HAGOs and HIOs and also allowed for the successful generation of pancreatic tissue indicating unambiguously that the PDX1 mutation is responsible for these pathologies. Based on these findings, these PDX1-null patients will have additional screening to monitor the progression of metaplasia into gastric and intestinal cancer. This study further demonstrates the utility of organoids in improving patient diagnoses and treatment.

Disclosure

M. Krishnamurthy: None. T.R. Broda: None. D.O. Kechele: None. X. Zhang: None. J.J. Palermo: None. M.H. Collins: None. I.H. Thomas: None. A. Heider: None. A. Dauber: None. J. Wells: None.

Funding

American Diabetes Association (1-19-PDF-036 to M.K.)
26-LB: Exenatide and Renal Outcomes in Patients with Type 2 Diabetes and Diabetic Kidney Disease: A Multicenter, Randomized, Parallel Study
Authors: WANG XIANGYU , ZHANG QIAN , GUAN MEIPING , SHENG SHUYUE , MO WEI , ZOU MENGCHEN , LI JIMIN , BI JIANLU , TANG XIANYU , HE JIALI , XU GUGEN , LI PING , ZHANG HUIJIE , XUE YAOMING
Published: June 2020

Aim: This trial aimed to assess the effects of exenatide, a GLP-1RA, on renal outcomes in patients with T2DM and DKD.

Methods: We performed a randomized, parallel study conducted in 4 general hospitals. T2DM patients with an eGFR≥30mL/min/1.73m2 and macroalbuminuria, defined as 24h-urinary albumin-creatinine ratio (UAER) >0.3g/24h, were randomized 1:1 to receive exenatide twice daily plus glargine or lispro insulin plus glargine for 24 weeks. The primary outcome was percentage change in 24h-UAER after 24 weeks of intervention comparing to baseline measurement. The secondary outcomes included the percentage change in urinary ACR, change in 24h-UAER, ACR, HbA1c, FPG, body weight and blood pressure. The safety outcomes include rates of hypoglycemia, adverse events and change in eGFR during the follow up.

Results: Between March 2016 and April 2019, 92 patients were randomized and took at least one dose of study drug. The mean age of the participants was 56 years. At baseline, the median 24h-UAER was 1512.0 mg/24h (IQR 782.5-2818.0), and mean eGFR was 70.4 mL/min/1.73 m2 (SD 23.4). After 24 weeks, exenatide reduced the percentage change of the 24h-UAER (difference between groups was 29.7%, p=0.0255), the percentage change of ACR (difference between groups was 28.4%, p=0.0395) and the absolute change of ACR (difference between groups was 607.8 mg/g, p=0.0209) but had no significant difference on the absolute change of 24h-UAER than the control group. Meanwhile, the body weight declined by 1.3kg with exenatide (difference between groups was 2.7kg, p=0.0001). There was no difference between two groups on the change in HbA1c, FPG, blood pressure or eGFR. Comparing to the control group, lower frequency of hypoglycemia as well as more gastrointestinal adverse events were in intervention group.

Conclusions: Exenatide twice daily plus glargine for 24 weeks resulted in significant reduction of albuminuria in T2DM patients with DKD.

Disclosure

X. Wang: None. Q. Zhang: None. M. Guan: None. S. Sheng: None. W. Mo: None. M. Zou: None. J. Li: None. J. Bi: None. X. Tang: None. J. He: None. G. Xu: None. P. Li: None. H. Zhang: None. Y. Xue: None.
261-OR: CDK4-R24C Compensates for IRS2 Deficiency by Enhancing Beta-Cell Proliferation and Restoring Signaling through the AKT-Foxo1-PDX1 Pathway
Authors: STAMATERIS RACHEL E. , SHARMA ROHIT B. , GABLASKI BRIAN , RANE SUSHIL G. , ALONSO LAURA C.
Published: June 2020

Expanding functional beta cell mass remains a prime goal of diabetes research. We previously demonstrated that IRS2 deficient mice have reduced islet expression of cyclin D2, and that overexpressing cyclin D2 rescues proliferation in IRS2 deficient beta cells in vitro. Since cyclin D2 partners with CDK4 to drive cell cycle progression, we hypothesized that an activated form of CDK4, CDK4-R24C (resistant to inhibition by Ink4A cell cycle inhibitors) might rescue the in vivo proliferation defect in IRS2 deficient mice. Mice heterozygous for both whole body Irs2 deletion and whole body Cdk4-R24C knock-in were mated, yielding 6 genotypes of interest: Irs2+/+; Cdk4-WT, Irs2+/+; Cdk4-R24C/WT, Irs2+/+; Cdk4-R24C/R24C, Irs2-/-; Cdk4-WT, Irs2-/-; Cdk4-R24C/WT, Irs2-/-; Cdk4-R24C/R24C. Two copies, but not one, of Cdk4-R24C completely rescued fasting and non-fasting blood glucose, glucose intolerance, and beta cell mass and proliferation, without rescuing peripheral insulin resistance. Interestingly, Irs2-/-; Cdk4-WT islets contained dedifferentiated (reduced PDX1, nuclear FOXO1, ALDH1A3+) beta cells, and this was completely reversed in Irs2-/-; Cdk4-R24C/R24C islets. An in vitro primary beta cell model of increased nuclear FOXO1 activity (short-term starvation) resulted in mRNA changes consistent with nuclear FOXO1 activity (including reduced Pdx1) and mimicked IRS2 deficiency in vivo. Intriguingly, overexpressing cell cycle activators CDK4 and cyclin D2 partially rescued the mRNA changes indicative of nuclear FOXO1 activity, while expressing an inhibitor (CDKN2A/p16) did not. Overexpression of CDK4 increased FOXO1 phosphorylation. We conclude that CDK4 may play additional roles outside of the cell cycle in beta cells, by activating the insulin signaling pathway downstream of IRS2 and exerting antidiabetic action by maintaining beta cell differentiation.

Disclosure

R.E. Stamateris: None. R.B. Sharma: None. B. Gablaski: None. S.G. Rane: None. L.C. Alonso: Consultant; Self; Fairbanks Pharmaceuticals Inc. Research Support; Self; American Diabetes Association.

Funding

American Diabetes Association (1-18-IBS-233 to L.C.A.)
265-OR: Identifying Adults at Risk of Unintentional Severe Hypoglycemia in Hospital Using Artificial Intelligence (RUSHH-AI)
Authors: FRALICK MICHAEL , DAI DAVID , POU-PROM CHLOE , VERMA AMOL A. , MAMDANI MUHAMMAD
Published: June 2020

Background: Machine learning carries great promise to improve healthcare delivery. Clinical outcomes that are routinely and objectively measured, and have serious consequences that can be prevented, are ideal targets for prediction and intervention. Hypoglycemia, defined as a blood glucose less than 3.9 mmol/L (70 mg/dL), meets these criteria. The purpose of this study was to predict hypoglycemia using artificial intelligence models in patients hospitalized to general internal medicine (GIM) and cardiovascular surgery (CV) at a tertiary-care teaching hospital in Toronto, Ontario.

Methods: Models were built using routinely-collected clinical data from the hospital’s electronic health record. Models were trained using data from Jan 2013-Apr 2017, tested using data from Apr 2017-Mar 2018, and validated using held-out test data from Apr 2018-Mar 2019. Three models were generated using supervised machine learning: LASSO regression, gradient boosted trees, and a recurrent neural network. Each model included baseline patient data and time-varying data. Natural language processing was used to incorporate text data from physician and nursing notes.

Results: We included 8492 GIM admissions and 8044 CV admissions. The average age of patients was 68 years, 35% were women, the baseline creatinine was 90 μmol/L (1.0mg/dL) and the baseline A1C was 7%. Hypoglycemia occurred in 15% of GIM admissions and 13% of CV admissions. The area under the curve for the model in the held-out validation set was approximately 0.80 on the GIM ward and 0.82 on the CV ward. When the threshold for hypoglycemia was lowered to 2.9 mmol/L (52 mg/dL), similar results were observed. Among the patients at the highest decile of risk, the positive predictive value was approximately 50% and the sensitivity was 99%.

Conclusion: Using natural language processing and machine learning we were able to accurately identify patients at high risk of hypoglycemia in hospital.

Disclosure

M. Fralick: None. D. Dai: None. C. Pou-Prom: None. A.A. Verma: None. M. Mamdani: None.
273-OR: Comparative Effectiveness of Lifestyle Intervention on Fasting Plasma Glucose in Normal Weight vs. Overweight/Obese Adults with Prediabetes
Authors: SALMON MANDY K. , WRIGHT BRENDA S. , KRIDL TERRI L. , FAIRCLOTH GEORGE C. , SALMON RICHARD D. , GORDON NEIL F.
Published: June 2020

Almost 1 in 5 U.S. adults with prediabetes have a normal BMI. It is hypothesized that such individuals develop diabetes through different pathophysiological mechanisms and that methods of prevention may differ in the absence of overweight/obesity. Normal BMI is an exclusion criterion for many diabetes prevention programs and scarce data are available on the effectiveness of lifestyle intervention in normal weight adults with prediabetes. In this study, we compared the effect of technology-enabled lifestyle health coaching (LHC) on fasting plasma glucose (FPG) in normal weight (BMI 18.5-24.9 kg/m2; n = 188), overweight (BMI 25.0-29.9 kg/m2; n = 495) and obese (BMI >30.0 kg/m2; n = 1,214) adults with prediabetes. Subjects were 1,897 consecutive adults with prediabetes (FPG = 100-125 mg/dl and not taking antidiabetic medication) who completed a baseline and follow-up evaluation as part of an LHC program (mean follow-up = 145 days). LHC included 1-on-1 behaviorally-oriented coaching, predominantly via the telephone and Internet, on exercise, nutrition and weight management. BMI decreased significantly (p <0.001) in the overweight (-0.6±1.1 kg/m2) and obese (-1.3±1.8 kg/m2) participants but was unchanged (0.0±1.5 kg/m2) in the normal weight participants. In contrast, weekly aerobic exercise increased (normal weight = 77±167 min/week increase; overweight = 98±191 min/week increase; and obese = 95±146 min/week increase) and FPG decreased (normal weight = -9±9 mg/dl decrease; overweight = -7±9 mg/dl decrease; and obese = -6±10 mg/dl decrease) significantly (p <0.001) in all 3 groups (p <0.05 for magnitude of decrease in FPG in normal weight versus obese participants). FPG was reduced to <100 mg/dl in 62.4%, 51.7% and 43.6% of the normal weight, overweight and obese participants, respectively. These data suggest that LHC is at least as effective in managing FPG in normal weight adults with prediabetes versus those who are overweight/obese.

Disclosure

M.K. Salmon: Other Relationship; Self; INTERVENT, International, LLC. B.S. Wright: Stock/Shareholder; Self; INTERVENT International, LLC. T.L. Kridl: Employee; Self; INTERVENT International, LLC. G.C. Faircloth: Stock/Shareholder; Self; INTERVENT International, LLC. R.D. Salmon: Stock/Shareholder; Self; INTERVENT International, LLC. N.F. Gordon: Board Member; Self; PrevCan. Stock/Shareholder; Self; INTERVENT International.
279-OR: Cognition in Young Adults with Youth-Onset Type 1 and Type 2 Diabetes: The SEARCH for Diabetes in Youth Study
Authors: SHAPIRO ALLISON , DABELEA DANA , STAFFORD JEANETTE M. , DAGOSTINO RALPH , PIHOKER CATHERINE , LIESE ANGELA D. , SHAH AMY S. , BELLATORRE ANNA , LAWRENCE JEAN M. , HENKIN LEORA , WILKENING GRETA
Published: June 2020

Poor cognition has been observed in children and adolescents with type 1 (T1D) and type 2 (T2D) diabetes. Differences in cognition between young adults with youth-onset T1D and T2D, however, have not been explored, nor is the effect of shared etiologic processes, e.g., poor glycemic control, on cognition fully understood. Using data from SEARCH for Diabetes in Youth, a multicenter, observational cohort study, we compared cognition between young adults with youth-onset T1D and T2D, testing whether glycemic control modified the relationship between diabetes type and cognition. Cognition was assessed via the NIH Toolbox Cognition Battery and age-corrected composite Fluid Cognition scores used. Glycemic control was estimated from repeated measures of HbA1c. Linear regression modeling was used to test the interaction effect of diabetes type and glycemic control on Fluid Cognition score while controlling for sex, race/ethnicity, waist-height ratio, duration of diabetes, depression, alcohol and drug use, any hypoglycemic episode in the past year, parental highest level of education, household income, and clinic site. The T2D group (n = 254) had significantly lower Fluid Cognition scores, compared to the T1D group (n = 953) (mean [SD] 84.7 [17.3] vs. 95.4 [16.8], p<0.001). Glycemic control modified the relationship between diabetes type and Fluid Cognition scores (interaction p=0.16). Using the model’s interaction estimates, at a HbA1c of 6.5%, the T2D group demonstrated significantly lower Fluid Cognition scores, compared to the T1D group (β = -6.54, 95% CI: -11.52, -1.57, p=0.01). At a HbA1c of 9.5%, Fluid Cognition scores in the T2D group differed to a lesser extent from the T1D group (β = -3.61, 95% CI: -7.16, -0.06, p=0.05). Youth with T2D had poorer cognition then youth with T1D. This effect is more pronounced at lower HbA1c levels. Our findings suggest that factors unique to youth-onset T2D may impact cognition in this specific diabetes group, warranting further investigation.

Disclosure

A. Shapiro: None. D. Dabelea: None. J.M. Stafford: None. R. Dagostino: Consultant; Self; Amgen, AstraZeneca, Celgene, Daiichi Sankyo. C. Pihoker: None. A.D. Liese: None. A.S. Shah: None. A. Bellatorre: None. J.M. Lawrence: None. L. Henkin: None. G. Wilkening: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (1UC4DK108173-01)
282-OR: The Association of Age of Onset of Type 2 Diabetes with Long-Term Risk of End Stage Kidney Disease: A National Registry Study
Authors: MORTON JEDIDIAH I. , LIEW DANNY , MCDONALD STEPHEN , SHAW JONATHAN E. , MAGLIANO DIANNA J.
Published: June 2020

This study aimed to estimate the long-term risk of End-Stage Kidney Disease (ESKD) in type 2 diabetes. We included 1,113,201 people with type 2 diabetes registered on the Australian National Diabetes Services Scheme (NDSS) followed from 2002 until 2013. The NDSS was linked to the Australia and New Zealand Dialysis and Transplant Registry and the National Death Index to obtain incident renal replacement therapy (RRT) and death, respectively. Between 2002 and 2013, 7,592 NDSS registrants initiated RRT over 7,839,075 person-years of follow up. The incidence of RRT increased with increasing duration of diabetes. In the first 10-15 years after onset of diabetes, incidence of RRT was highest in those with older age at onset of diabetes. However, over longer durations of diabetes, the cumulative incidence of RRT became higher in those with younger onset diabetes. When “total” incidence of ESKD (RRT and deaths attributed to ESKD without RRT) was considered, the predicted lifetime risks of renal failure (until age 100) were 20.1%, 16.1%, 11.6%, 7.7%, 5.4%, and 4.2% in those with diabetes onset between ages 10-29, 30-39, 40-49, 50-59, 60-69, and 70-79, respectively. Thus, the long-term risk of ESKD in type 2 diabetes is high, which disproportionately affects those with younger-onset type 2 diabetes as they are more likely to survive to longer diabetes durations.

Disclosure

J.I. Morton: None. D. Liew: None. S. McDonald: None. J.E. Shaw: Advisory Panel; Self; AstraZeneca, Merck Sharp & Dohme Corp., Mylan, Sanofi. Research Support; Self; AstraZeneca. Speaker’s Bureau; Self; Eli Lilly and Company, Mylan. D.J. Magliano: None.

Funding

Australian Government Research Training Program (to J.I.M.); Monash University (to J.I.M.)
284-OR: Characterization of Cognitive Changes in Type 1 Diabetes of Extreme Duration by Retinal Neural Structure and Vasculature
Authors: WOLFSON EMILY , FICKWEILER WARD , SAMPANI KONSTANTINA , SHAH HETAL , AIELLO LLOYD P. , SUN JENNIFER , KING GEORGE L.
Published: June 2020

The risk of cognitive dysfunction in aging populations with type 1 diabetes (T1D) may be similar to that of people with type 2 diabetes, but it has not been well-characterized clinically. In the Joslin Medalist Study, a cohort of people with ≥ 50 years of T1D, cognitive dysfunction was previously shown to correlate with cardiovascular disease (CVD) and proliferative diabetic retinopathy (PDR). In a Medalist subset (n=129), we extended these findings by assessing the relationships of cognitive changes and other complications with vascular abnormalities in the retina, a developmental extension of the central nervous system. Medalists were evaluated with validated cognitive tests to assess psychomotor speed (PS), immediate memory (IM), delayed memory (DM), working memory (WM), and executive function (EF). Retinal optical coherence tomography (OCT) and OCT angiography were performed to obtain neural retinal layer thickness and vascular density for the superficial (SCP) and deep (DCP) retinal capillary plexuses. CVD and PDR were associated with worse IM (p=0.03) and decline in PS (p=0.01), respectively, adjusting for age, A1c, BMI, triglycerides and hypertension. Thinning of the retinal outer nuclear layer (ONL) was associated with PDR (p=0.04), CVD (p=0.02), worse IM (p=0.05) and PS (p=0.01), adjusted for sex, diabetes duration, PDR and visual acuity. CVD, PDR and DM severity were associated with decreased DCP density (p=0.04, p=0.0004, and p=0.002, respectively). Decreased density of the SCP was associated with worse dominant hand PS (p=0.01), adjusted for PDR, T1D duration, and visual acuity.

In summary, cognitive decline in people with long duration of T1D is associated with PDR and CVD, which is supported by shared specific retinal vascular imaging abnormalities, suggesting a potential common pathogenesis underlying these complications in T1D.

Disclosure

E. Wolfson: None. W. Fickweiler: None. K. Sampani: None. H. Shah: None. L.P. Aiello: Advisory Panel; Self; Novo Nordisk Inc. Consultant; Self; KalVista Pharmaceuticals, Inc., Novo Nordisk Inc. Stock/Shareholder; Self; KalVista Pharmaceuticals, Inc. Other Relationship; Self; Optos. J. Sun: Research Support; Self; Adaptive Sensory Technology, Boehringer Ingelheim Pharmaceuticals, Inc., KalVista Pharmaceuticals, Inc., Novo Nordisk Inc., Optovue, Inc., Roche Pharma. Other Relationship; Self; Merck Foundation, Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Roche Pharma. G.L. King: Research Support; Self; Janssen Pharmaceuticals, Inc.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (P30DK036836, UL1RR025758-03, R24283, DK083957-01DP3, DK094333-01, T32DK007260); JDRF (17-2013-310)
286-OR: Biomedical Predictors of Cognitive Decline in Older Participants with T1DM: Longitudinal Follow-Up of the DCCT/EDIC Cohort
Authors: JACOBSON ALAN M. , RYAN CHRISTOPHER M. , BRAFFETT BARBARA H. , GUBITOSI-KLUG ROSE , LORENZI GAYLE M. , LUCHSINGER JOSE A. , TRAPANI VICTORIA R. , BEBU IONUT , CHAYTOR NAOMI , HITT SUSAN M. , FARRELL KALEIGH , LACHIN JOHN M.
Published: June 2020

DCCT/EDIC participants have been followed for an average of 32 years (N=1,054; mean age 60±7 years; duration T1DM 38±5 years). To evaluate cognitive function in T1DM, mental efficiency and memory were assessed five times during the study. Standardized z-scores were calculated relative to the DCCT baseline evaluation and averaged to obtain three summary domain scores (immediate recall, delayed recall, mental efficiency). Generalized linear mixed models were used. In 2018, mean HbA1c levels were 7.9±0.9% and 46% of participants had experienced ≥1 hypoglycemic coma/seizure. Adjusted for age, sex, and education, cognition declined over time (p<0.0001 each domain). In multivariable analysis, higher mean HbA1c (β±SE, -0.105±0.017 per 1%, p<0.0001), higher mean systolic blood pressure (-0.083±0.013 per 5 mmHg, p<0.0001), and history of coma/seizure (-0.175±0.041 ≥1 vs. none, p<0.0001) were independently associated with declining mental efficiency. Decreased mental efficiency was also associated with microalbuminuria (AER ≥30 mg/hr), lower levels of HDL cholesterol, and higher daily insulin dose (u/kg/day) (each p<0.0001). Biomedical predictors did not influence recall. These findings demonstrate the effects of increasing age, glycemic excursions (chronic hyperglycemia and clinically-significant hypoglycemia), and blood pressure on mental efficiency.

Disclosure

A.M. Jacobson: None. C.M. Ryan: None. B.H. Braffett: None. R. Gubitosi-Klug: None. G.M. Lorenzi: None. J.A. Luchsinger: Consultant; Self; vTv Therapeutics. V.R. Trapani: None. I. Bebu: None. N. Chaytor: Consultant; Self; Eli Lilly and Company. S.M. Hitt: None. K. Farrell: Stock/Shareholder; Spouse/Partner; Dexcom, Inc., Tandem Diabetes Care. J.M. Lachin: Board Member; Self; Tolerion, Inc.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases
287-OR: TCR/HLA Humanized Mice Reveal Reduced Tolerance and Increased Immunogenicity of Posttranslationally Modified GAD65 Epitope
Authors: JING YI , KONG YUELIN , MCGINTY JOHN , BLAHNIK-FAGAN GABRIELE , BETTINI MATTHEW , JAMES EDDIE A. , BETTINI MARIA
Published: June 2020

T cells specific for post-translationally modified (PTM) islet antigens have been implicated as critical players in the development of type 1 diabetes. T cells reactive to the deamidated version of glutamic acid decarboxylase 65 (GAD65) 115-127 epitope accumulate in peripheral blood of patients as they progress to disease. However, knowledge regarding the development, activation, differentiation and islet recruitment of PTM reactive T cells is limited. Using retroviral gene delivery, we have developed a humanized TCR/HLA mouse model to assess the in vivo function of human TCRs reactive to either the wild type (GADWT) or the deamidated (GAD120E) GAD65115-127 epitope. Both TCRs supported efficient thymic selection; however, fewer regulatory T cells developed in mice expressing the PTM reactive TCR, which was partially rescued by the thymic delivery of GAD120E peptide. Immunization of immune sufficient polyclonal DR4.NOD mice with GAD120E/CFA elicited a stronger response compared to GADWT/CFA, confirming that tolerance to the wild type autoantigen is more robust. Interestingly, while islet infiltration in GAD120E single TCR mice was dependent on priming with the cognate PTM peptide, GADWT-TCR T cells could be induced to infiltrate islets after priming with either GADWT or GAD120E peptide, despite the vastly different functional affinity (EC50, GADWT: 0.047μM vs. GAD120E: ≥1.4μM). These findings suggest that the development and expansion of GAD65 PTM reactive CD4+ T cells are less restricted in peripheral lymphoid organs, but their recruitment to the tissue site is dependent on the availability of PTM antigen.

Disclosure

Y. Jing: None. Y. Kong: None. J. McGinty: None. G. Blahnik-Fagan: None. M. Bettini: None. E.A. James: None. M. Bettini: None.

Funding

National Institutes of Health (R01AI125301)