bluuebunny/crossref_metadata_2025 huggingface dataset:
1700-P: Optimizing Data Cleaning Methods for Remotely Collected Electronic Weight Measurements
Authors: DAREDIA SAHER , SMITH BRIDGETTE P. , SERRATO BANDERA HILLARY , MILLMAN ANDREA , QUESENBERRY CHARLES , FERRARA ASSIAMIRA , BROWN SUSAN D.
Published: June 2023

Background: Digital smart scales transmit weight measurements collected outside of clinical settings to medical and research teams. Statistical packages such as SAS %TPF (TrueProfileFinder) can ensure weights belong to persons of interest instead of others who may use the scale; however, validation is required.

Methods: We obtained 32,917 remotely collected weights from 222 postpartum participants using BodyTrace scales in the Pregnancy, Environment, and Lifestyle Study-2 (PETALS-2). To assess the SAS %TPF macro's performance, we applied 18 combinations of its three modifiable parameters (number of observations in a moving window, importance of weights from clinic visits, and prediction band) to a random subsample (N=16). In parallel, two manual reviewers independently classified weights as ‘belonging’ or ‘not belonging’ to participants; Cohen's κ ≥0.91 confirmed adequate interrater agreement. Outputs from the 18 models were then compared to the manual review.

Results: As hypothesized, adjusting parameters impacted the macro's ability to identify weights truly belonging to participants (percent true calls). Prediction band, the predicted range for new observations based on a linear regression model, was the only parameter associated with macro performance. A prediction band of 15 lbs. was associated with a 2.71% (95% CI: 0.16%, 5.25%) increase in percent true call and 5.57% (95% CI: 1.77%, 9.38%) increase in sensitivity vs. a band of 10 lbs. Percent true call for a prediction band of 15 lbs. was 2.95% higher, but non-significant (p=0.18) vs. a band of 20 lbs. A prediction band of 15 lbs. was highly accurate (>97% true call) and sensitive (>98%).

Conclusion: Weight is often assessed in interventions for glycemic control and diabetes prevention. Precise data cleaning can increase accuracy of unsupervised, remotely collected measurements. Clinicians and researchers using smart scales can apply this data cleaning method to identify SAS %TPF macro parameters that maximize accuracy.

Disclosure

S.Daredia: None. B.P.Smith: None. H.Serrato bandera: None. A.Millman: None. C.Quesenberry: None. A.Ferrara: None. S.D.Brown: None.

Funding

National Institutes of Health (R01HL142996)
1702-P: Altered Gene Expression Signatures following COVID-19 Infections and/or mRNA Vaccination in Human Adipose Tissue
Authors: KRUPKA SONTJE , BLÜHER MATTHIAS , KLÖTING NORA
Published: June 2023

Obesity is associated with an increase in morbidity and mortality from Coronavirus disease- 2019 (COVID-19). Adipose tissue (AT) demonstrates high expression of angiotensin-converting enzyme 2 (ACE2), a host cell entry factor of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 has been shown to infect AT, where it continues to proliferate, thereby contributing to local inflammation. Since visceral AT inflammation may play a role in the pathogenesis of type 2 diabetes, we investigated the molecular and histological AT signatures after a Covid-19 infection and/or mRNA vaccination. We investigated 54 paired subcutaneous (SC) and visceral adipose tissue (VAT) biopsies from the Leipzig Obesity BioBank in subgroups based on positive versus negative SARS-CoV-2 infection as well as mRNA vaccination status in comparison to BMI and age matched controls obtained before appearance of Covid-19 (N=27). We compared gene expression signatures by RNA-Sequencing analyses, morphology changes in AT as well as circulating inflammation markers across the groups. In relation to a Covid-19 infection, we find a distinct gene expression signature and morphology in AT suggesting that SARS-CoV-2 affects AT and may mediate the disease course. In this context, inflammation of AT, but also an altered secretion profile from AT could link obesity to its metabolic complications, but also to long Covid-19 disease courses.

Disclosure

S.Krupka: None. M.Blüher: Advisory Panel; Boehringer Ingelheim Inc., Lilly, Novo Nordisk, Consultant; Novo Nordisk Foundation, Speaker's Bureau; Amgen Inc., AstraZeneca, Bayer Inc., Daiichi Sankyo, Novartis, Sanofi-Aventis Deutschland GmbH. N.Klöting: None.
1704-P: Efficacy and Safety of Tirzepatide for Weight Loss—A Meta-analysis of Randomized Controlled Trials
Authors: MESQUITA YASMIN L.L. , CALVI IZABELA P. , MARQUES ISABELA R. , CRUZ SARA A. , PADRAO EDUARDO M.H. , CARDOSO RHANDERSON , MOURA FILIPE , RAFALSKIY VLADIMIR V.
Published: June 2023

Tirzepatide (TZP) is a new dual glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1 receptor agonist approved for type 2 diabetes and shown to have substantial efficacy on weight loss. We performed a meta-analysis to assess the efficacy and safety of TZP for weight loss. We searched PubMed, Cochrane and Embase up until July 2022 for RCTs comparing TZP with placebo for the co-primary endpoints of absolute and percent change in weight. Mean difference (MD) and odds ratios (OR) were utilized, respectively, for continuous and binary outcomes. Of 397 search results, 6 studies were included, with 4036 participants. Pooled analysis showed that TZP 5 mg, 10 mg, and 15 mg were more effective than placebo, with MD in body weight change ranging from -7.7 kg (95% CI -11.0, -4.4; p < 0.001) to -11.8 kg (95% CI -17.4, -6.2; p < 0.001) - Figure 1A. The MD in percent change in weight ranged from -8.1% (95% CI -11.0, -5.2; p < 0.001) to -12.4% (95% CI -17.2, -7.5; p < 0.001) - Figure 1B. Adverse events were more frequent with TZP compared with placebo, especially with the 15 mg dose, with respect to nausea (OR 4.2; 95% CI 2.4, 7.5; p < 0.001), vomiting (OR 7.0; 95% CI 4.3, 11.4; p < 0.001), and diarrhea (OR 2.8; 95% CI 1.6, 4.9; p < 0.001). The results support that TZP leads to a significant reduction in body weight and may be a valuable therapeutic option for weight management, despite an increase in gastrointestinal side effects.

Disclosure

Y.L.L.Mesquita: None. I.P.Calvi: None. I.R.Marques: None. S.A.Cruz: None. E.M.H.Padrao: None. R.Cardoso: None. F.Moura: None. V.V.Rafalskiy: None.
171-OR: Diagnosing GDM in Pregnancy—Closing the Door after the Horse Has Bolted?
Authors: YAJNIK CHITTARANJAN S. , BANDYOPADHYAY SOUVIK , BHALERAO ABOLI A. , BHAT DATTATREY , PHATAK SANAT , WAGH RUCHA H. , YAJNIK PALLAVI C. , OTIV SUHAS R. , COYAJI KURUS J. , FALL CAROLINE
Published: June 2023

RCTs of GDM treatment reduces adverse pregnancy outcomes but not long-term offspring diabesity which is influenced by genetic and epigenetic programming. The vital window for programming is pre- and immediate post-conceptional, strongly influenced by maternal nutrition and metabolism. Diagnosing GDM later in the pregnancy systematically misses this window. Metabolic-endocrine abnormalities of ‘GDM’ are detectable years before pregnancy (Catalano 2013; Han 2016). There is little data to track the life course evolution of pregnancy hyperglycaemia. Pune Maternal Nutrition Study (1993) has serial glycemic and BMI data on girls born in the cohort at 6, 12 and 18 years, in pregnancy and post-delivery. By 2020, pregnancy data (28-wks gestation) was available on 170. They were 21-years old, BMI 22 kg/m2, twenty had GDM (IADPSG). We compared serial data of Q4 of fasting plasma glucose (FPG, N=44, ‘hyperglycemic’) with 126 ‘normoglycemic’. Figure shows that hyperglycemic girls had higher FPG and BMI at 6, 12 and 18-years and continued to remain so post-delivery. Our data shows that pregnancy glycemia reflects life course glycemia. Diagnosing ‘GDM’ in pregnancy ignores exposure of ova and young conceptus to an abnormal metabolic milieu, failing to prevent periconceptional programming of diabesity. There is an urgent need to reconsider strategies about GDM diagnosis and treatment to curtail the escalating epidemic of diabesity in the young.

Disclosure

C.S.Yajnik: None. C.Fall: None. S.Bandyopadhyay: None. A.A.Bhalerao: None. D.Bhat: None. S.Phatak: None. R.H.Wagh: None. P.C.Yajnik: None. S.R.Otiv: None. K.J.Coyaji: None.

Funding

UK Wellcome Trust (038128/Z/93, 059609/Z/99, 079877/Z/06/Z, 098575/B/12/Z, 083460/Z/07/Z); Medical Research Council, UK (MR/J000094/1); Department of Biotechnology, Government of India (BT/PR-6870/PID/20/268/2005); KEM Hospital Research Centre
1712-P: Serum miRNA-1 Levels as a Potential Marker for Myocardial Steatosis in Normal Weight and Obese/Overweight Diabetic Individuals—A Pilot Study
Authors: PALANISAMY SRIKANTH , PARLATAN UGUR , OZEN MEHMET , KARIM ASMA H. , AKIN DEMIR , DEMIRCI UTKAN , PALANIAPPAN LATHA
Published: June 2023

Of the 37 million Americans with type II diabetes, a substantial proportion have a normal body mass index (BMI). These populations suffer a disproportionately high burden of mortality; the development of diabetic cardiomyopathy (DCM) is likely a contributor. Myocardial steatosis, a known consequence of diabetes, is a significant part of DCM's pathogenesis; unfortunately, there is no feasible way to screen patients for steatosis. Exosomal microRNAs (miRNAs) are a promising biomarker for this purpose. However, no study has evaluated miRNA signatures in normal-weight patients with diabetes. Serum samples of obese/overweight and normal-weight diabetic subjects were obtained via IMPACT and STRONG-D clinical trial protocols respectively. Exosomes were extracted using the ExoTIC (exosome total isolation chip) platform. MiRNA-1 levels from extracellular vesicles were quantified using RT-qPCR and compared via unpaired student's two-tailed t-test. A total of 8 samples (4 normal-weight and 4 overweight/obese) were analyzed. Overall, the obese cohort had a mean age of 63.7 vs. 57.2 for the normal-weight cohort. The obese cohort had a mean BMI of 29.7 vs. 23.1 for the normal weight cohort. Both cohorts were composed of 75% men and 25% women. The obese cohort had a mean hemoglobin A1c of 7.2% vs. 8.0% in the normal-weight cohort. Serum miRNA-1 levels were significantly higher in normal-weight diabetic subjects (p = 0.0004). In this pilot study, patients with diabetes of a normal weight had significantly higher levels of serum miRNA-1, implying that they may have more myocardial steatosis and possibly, more cardiac dysfunction. This pathophysiologic difference could help explain the significant morbidity and mortality that diabetic patients of a normal weight suffer. Further directions for our study include the complete quantification of both miRNA-1 and miRNA-133a among our entire cohort, as well as assessment of echocardiographic parameters.

Disclosure

S.Palanisamy: None. U.Parlatan: None. M.Ozen: None. A.H.Karim: None. D.Akin: None. U.Demirci: None. L.Palaniappan: None.

Funding

National Institutes of Health (1R18DK096394-01A1, 2R01DK081371-06A1)
1716-P: Real-World Outcomes Using Medication and Intensive Lifestyle Intervention to Treat Obesity and Improve Diabetes at Scale
Authors: BOYD KIM , MEDEIROS KAELEN L. , KRAMER DANIEL , CELIO CHRISTINE , RYAN DONNA H. , KENYON ISABELLE
Published: June 2023

Introduction: Overweight and obesity impact >200 million Americans, driving rates of diabetes and cardiometabolic disease. To address this, we established a digital metabolic health company to provide medically-supervised obesity treatment at scale.

Methods: Treatment: GLP-1 receptor agonist medications combined with a proprietary intensive lifestyle intervention. Data analysis: Individuals who reached 12 months in the core program and a subset who continued to 18 months were analyzed for weight change, metabolic labs (baseline and 12mo) and patient-completed questionnaires (baseline, 6, and 12mo).

Results: The cohort included 2,643 patients. At baseline, 3.9% had type 2 diabetes, and 22.8% had prediabetes. Average weight loss at 12, 15 and 18 months was 15.58% (n=2,397), 15.97% (n=1,319) and 15.82% (n=569). Average weight loss for patients with diabetes or prediabetes at 12, 15 and 18 mo was 16.22% (n=299), 16.60% (n=233), and 15.57% (n=133). Patients with diabetes saw average improvements of -1.42 percentage points for HbA1c and -14.05 units for insulin; for patients with prediabetes, these were -0.45 reduction to HbA1c and -7.05 for insulin. 72.9% of patients with abnormal HbA1c levels at baseline were normal at 12 months. Positive changes were also seen for other metabolic markers and for patient-reported outcomes.

Discussion: Results from 2,643 enrollees in a new digital metabolic health program demonstrate robust and sustained weight loss through 18 months. For those with diabetes or prediabetes, there was significant weight loss, reduction in HbA1c and insulin with categorical improvement to diabetes status in 80.5%, improvements to other metabolic markers, and positive changes to health behaviors. These results offer great promise in addressing the obesity epidemic and reversing diabetes.

Disclosure

K.Boyd: Employee; Calibrate Health, Inc. K.L.Medeiros: Employee; Calibrate Inc. D.Kramer: Employee; Calibrate Health, Inc. C.Celio: None. D.H.Ryan: Advisory Panel; Altimmune, Wondr Health, YSOPIA, Zealand Pharma A/S, Consultant; Amgen Inc., Boehringer Ingelheim Inc., Carmot Therapeutics, Inc., Epitomee, Lilly, Other Relationship; Calibrate, Gila, IFA Celtic, Scientific Intake,, real appeal, Scientific Intake, Xeno Bioscience, Novo Nordisk. I.Kenyon: None.
1721-P: Patients with Metabolic Syndrome Are at Risk of Renal Outcome in CKD Stages 1-4 Regardless of Diabetes
Authors: LIN HUGO Y.H. , CHANG KAI-TING
Published: June 2023

Metabolic syndrome (MetS) is a cluster of risk factors predicting cariometabolic disease. MetS was associated with renal outcome in some studies. However, the effect of MetS seemed loss after the adjustment of proteinuria, late CKD stage or diabetes in advanced CKD. Whether MetS predicts renal outcome in established CKD is unknown. We investigated the prognostic effects of MetS in 2500 patients with CKD stage 1-4. 66.9% and 49.2% patients had MetS and DM, respectively. The MetS score was positively associated with proteinuria (p<0.001), inflammation (p=0.001), and nutrition markers (p<0.001). In fully-adjusted Cox regression model, hazard ratio (HR) (95% confidence interval) of MetS for composite renal outcome in DM and non-DM subgroup was 1.56 (1.15-2.12) (p<0.05) and 1.31 (1.02-1.70) (p<0.05), respectively, while that for all-cause mortality was 1.00 (0.71-1.40) and 1.27 (0.92-1.74). Blood pressure is the most important component of MetS for clinical outcomes (1.50 (1.08-2.08) (p<0.05)). In a 2 by 2 matrix, compared with non-MetS non-DM group, MetS DM group (HR: 1.62 (1.31-2.02) (p<0.05)) and MetS non-DM group (HR:1.33 (1.05-1.69) (p<0.05)) had higher risks for composite renal outcome, while MetS DM group had higher risk for all-cause mortality (HR: 1.43 (1.09-1.88) (p<0.05)). In conclusion, MetS could predict renal outcome in CKD stage 1-4 patients independent of diabetes.

Disclosure

H.Y.H.Lin: None. K.Chang: None.

Funding

Ministry of Science and Technology (110-2314-B-037-068-MY3); Municipal Ta-Tung Hospital (KMTTH-110-003)
173-LB: Circulating Calprotectin Is Increased in Children with T1D and Obesity—Implications for T1D Progression and Outcome
Authors: HASAN AMAL , KOCHUMON SHIHAB P. , ALBASIRI IMAN D. , ALBELOUSHI SHAIMA , ALOZAIRI EBAA S. , ALMULLA FAHD
Published: June 2023

Purpose: Circulating calprotectin is elevated in inflammatory disorders and is a biomarker of severity in inflammatory rheumatic and bowel diseases. Calprotectin has immunomodulatory properties that may influence the progression and outcome of inflammatory diseases. Calprotectin is elevated in children with obesity, but its levels in children with T1D co-morbid with obesity remain unexplored. We examined the levels of circulating calprotectin in children with T1D and obesity and compared these levels with those of children with normal weight as well as those with obesity and T2D or children with obesity only (no prediabetes or T2D).

Methods: Sixty-four children (aged 5-13 years) were included. Participants' characteristics included being of normal weight (body mass index, BMI <85th percentile; n = 12), having obesity (BMI ≥95th percentile) with T1D (n = 4), obesity with T2D (n = 8), or having obesity without prediabetes or diabetes (n = 40). Clinical markers were assessed using established procedures on peripheral blood. The levels of calprotectin were quantified using commercially available ELISA kits.

Results: Circulating calprotectin was significantly (P = 0.0044) higher in children with T1D and obesity, as well as in children with T2D and obesity (P = 0.02), compared to healthy children with normal weight. There was no difference in calprotectin levels between children with T1D and obesity compared to those with T2D and obesity or obesity without prediabetes or diabetes.

Conclusion: Increased calprotectin in children with T1D (as well as T2D) is due to the obese state and is not further increased by having diabetes. Given the immunomodulatory properties of calprotectin, obesity in children with T1D might hasten disease progression, resulting in a worse outcome. These findings highlight the need for tailored obesity prevention in children with high risk for T1D and emphasize the importance of treatment strategies for weight management in children with T1D.

Disclosure

A. Hasan: None. S. P. Kochumon: None. I. D. Albasiri: None. S. Albeloushi: None. E. S. Alozairi: None. F. Almulla: None.

Funding

Kuwait Foundation for the Advancement of Sciences
1735-P: The Dichotomic and Timely Regulation of SerpinB13 Plays a Role in Beta-Cell Neogenesis and Pancreatic Islet Expansion
Authors: KRYVALAP YURY , MENG SHAWN Z. , HABTE RIM , CZYZYK JAN
Published: June 2023

There has been recent emphasis on the potential role of a functional crosstalk between the exocrine and endocrine pancreas in the development of type 1 diabetes (T1D). Our interest in this was sparked by our observations that, (i) serpinB13, a protease inhibitor of cathepsin L, is expressed in mouse pancreatic ductal cells, and (ii) an antibody response to serpinB13 is associated with better clinical outcomes in mice and humans with T1D. The exact pattern of serpinB13 expression in the human pancreas has, to date, not been examined. Similarly, the exact role of serpinB13 in beta-cell biology and diabetes remains unclear. We aimed to determine the distribution of serpinB13 expression using immunochemistry on pancreatic sections obtained from nPOD. In addition, we developed a mouse serpinB13 genetic knockout model to examine the role of this serpin in regulating pancreatic islet morphology. Visiopharm software was used for an unbiased quantitative image analysis of islet size and cellularity. We found that in healthy humans, serpinB13 expression was confined to the pancreatic ducts, mimicking the pattern we observed previously in rodents. Studies of serpinB13 genetic deficiency revealed that early inhibition of serpinB13 increased the small pancreatic islet population while with time this trend reversed, and a drop in the number of pancreatic islets was observed. Our results demonstrate that serpinB13 is expressed in the exocrine epithelial compartment of the human pancreas. In addition, quite unexpectedly, serpinB13 appears to play a dual role in regulating pancreatic islet mass. More specifically, we postulate that serpinB13 initially hampers the development of additional insulin-producing cells, but with time it promotes expansion of the already formed beta cells. Thus, we propose an approach that first impedes serpinB13, followed by later administration of this serpin, which may lead to an increase in beta-cell mass and improved resistance to T1D.

Disclosure

Y.Kryvalap: None. S.Z.Meng: None. R.Habte: None. J.Czyzyk: None.
174-LB: Autoantibodies against PTM-IA2 Extracellular Domain Are Only Associated with Late Progression of Type 1 Diabetes
Authors: JIA XIAOFAN , WENZLAU JANET M. , REWERS MARIAN , MICHELS AARON W. , YU LIPING
Published: June 2023

Background: Autoantibodies directed against the extracellular domain of IA2 (IA2ecA) with post-translationally modified (PTM) (deamidated) epitopes have been identified in new-onset T1D patients. The prevalence and timing of appearance of PTM-IA2ecA in pre-T1D individuals is evaluated in the current study.

Methods: A PTM-IA2ec protein antigen probe containing four T-cell responsive deamidated Q>E peptides (aa 198-216, aa 467-482, aa 523-536 and aa 545-562) was produced. The study subjects included 269 children and 260 adults with new-onset T1D, 271 adults with LADA, 274 children with new-onset diabetes but negative for all islet autoantibodies (IAbs) (IAA, GADA, IA-2A and ZnT8A), and 222 pre-T1D children who were longitudinally followed from birth to stage 1-3 T1D. The longitudinal follow-up serum samples included samples prior to seroconversion, samples at the first appearance on an IAb and the last follow-up samples. All serum samples were tested for PTM-IA2ecA and the wide type (WT-IA2ecA) with a radio binding assay.

Results: In new-onset T1D, positivity for PTM-IA2ecA and WT-IA2ecA were 60.6% (163/269) and 19.3% (52/269), respectively in children, while significantly less in adults, 14.2% (37/260, P<0.0001) and 8.8% (23/260, P=0.0007), respectively. The positivity for PTM-IA2ecA and WT-IA2ecA were only 4.1% (11/271) and 1.5% (4/271) in LADA patients, 1.5% (4/274) and 1.1% (3/274) in diabetic children who were otherwise IAb negative. In 222 longitudinally followed children with pre-T1D, only 13 were tested positive for PTM-IA2ecA and 8 positives for WT-IA2ecA, with most detected at diagnosis of stage 3 T1D.

Conclusion: Autoantibodies against either WT-IA2ec or PTM-IA2ec were not frequently seen in pre-T1D as shown in new onset T1D. These autoantibodies develop in the late stage of T1D progression with overt clinical disease.

Disclosure

X. Jia: None. J. M. Wenzlau: None. M. Rewers: Research Support; Provention Bio, Juvenile Diabetes Research Foundation (JDRF), Hemsley Charitable Trust, Dexcom, Inc., Janssen Research & Development, LLC. A. W. Michels: Stock/Shareholder; Immnomolecular Therapeutics. L. Yu: None.

Funding

National Institute of Allergy and Infectious Diseases (AI53665); National Institute of Diabetes and Digestive and Kidney Diseases (DK032083)
1746-P: The Role of Cell–Cell Junction Associated Mediator in Glucose-Stimulated Insulin Secretion of Pancreatic Islets
Authors: WANG HAO
Published: June 2023

Glucose-stimulated insulin secretion from β cells is essential in maintaining glucose homeostasis. Cell-cell communication within the pancreatic islets, mediated by cell-cell junctions and their mediators, regulates insulin secretion dynamics and glucose homeostasis. It has been reported that Nephrin-mediated intercellular junction between β cells is implicated in the regulation of insulin secretion, however, the underlying mechanisms are not fully understood. Here, we investigated the effects of Girdin, a cell-cell junction associated mediator, on insulin release. We found that Girdin was expressed in mouse pancreatic β cells and mediated glucose-stimulated insulin secretion, especially for the second phase. Next, we demonstrated that glucose-stimulated tyrosine phosphorylation of Girdin mediated Nephrin phosphorylation by recruiting Src kinase, which leads to the endocytosis of Nephrin. Furthermore, we observed the glucose-induced Girdin/Nephrin/Src signaling induced downstream Akt phosphorylation and activated Rac1-mediated cytoskeleton reorganization, allowing insulin secretory granules to access the plasma membrane for the second-phase secretion. Finally, we found that Girdin is downregulated in the diabetic islets, and rescue of Girdin restored impaired glucose-stimulated insulin secretion. Taken together, these data provide evidence that the cell-cell junction associated mediator play important roles in fusion of insulin secretory vesicles within the cell and establish a role for Girdin in regulating this process.

Disclosure

H.Wang: None.

Funding

National Natural Science Foundation of China
175-LB: Altered Glycogen Cell Number in Placental mTOR Signaling Deficiency Model in Mice
Authors: CLIFTON BRIANA , BEETCH MEGAN , ALEJANDRO EMILYN
Published: June 2023

Fetal Growth Restriction (FGR) is a determinant for the development of chronic diseases, such as type II diabetes in adulthood. FGR is caused by the failure of the placenta to provide adequate nutrients and oxygen for fetal development. mTOR regulates cell metabolism and growth, and it is reduced in the FGR placenta. We previously reported that placental mTOR ablation induces insulin resistance and glucose intolerance in adult mice. We hypothesize that mTOR signaling regulates glucose utilization in the placenta, which stores glucose reservoirs as glycogen. Appropriate localization and breakdown of glycogen cells is crucial for normal fetal growth. We leveraged our mouse model lacking placental mTOR signaling to ask whether glycogen cell number was altered. First, we found that critical glycogenesis and glycogenolysis enzymes, as well as the glucagon receptor, were expressed in placental tissue, indicating the capability for glycogen cell synthesis and breakdown, like glycogen-rich tissues such as the liver. Next, we show increased glycogen cell counts in the junctional zone of the mTORKO placenta compared to littermate controls by IHC. However, whole placental glycogen content, measured by ELISA, demonstrates no changes between KO and control. These data suggest that mTOR signaling modulates the localization and migration of glycogen cells during high metabolic needs. Glucagon mRNA is expressed in the placenta, with significant increases in the male KO placenta, indicating the placenta may produce its own glucagon. However, IHC staining indicates increased glucagon levels in female KO placenta in the labyrinth zone. Together, these data suggest that the placenta may have the capability to regulate its own glycogen breakdown by altering glucagon expression. Our results indicate that the FGR placenta induced by mTOR deletion has altered glycogen cell content and mislocalization, which mimics the phenotypes of placenta impacted by gestational diabetes, suggesting a shared mechanistic pathway.

Disclosure

B. Clifton: None. M. Beetch: None. E. Alejandro: None.

Funding

National Institutes of Health (R01DK115720)
175-OR: Pregnancy Intervention with a Closed-Loop System (PICLS) Study
Authors: BUSCHUR ELIZABETH , SNELL-BERGEON JANET K. , PYLE LAURA , GARCETTI RACHEL , DUNGAN KATHLEEN M. , THUNG STEPHEN F. , BARTHOLOMEW ANNA , SUMMERFIELD TARYN , BLOKS TANNER , NEASE EMILY R. , POLSKY SARIT
Published: June 2023

We performed a parallel, two-site, randomized controlled trial of a commercially available hybrid closed-loop (HCL) system compared to sensor augmented pump therapy (SAPT) in pregnant women with type 1 diabetes (T1D). Participants started SAPT (Medtronic 670G) by 11 weeks gestation, were randomized at 14-18 weeks, and continued therapy until 6 weeks postpartum. The HCL group used assistive techniques, “fake carbohydrate” boluses, to optimize therapy. We performed linear mixed model regression to compare outcomes between groups, adjusted for clinic. There were 24 women randomized and 23 completers. At baseline there were no significant differences between groups (mean baseline A1C 6.8% at 8w5d [SAPT]/9w0d [HCL] enrollment). There were no episodes of severe hypoglycemia or diabetic ketoacidosis after randomization. Adverse skin reactions occurred in 6 participants (5 SAPT vs 1 HCL, p=0.1550) with a median (IQR) number of 1 (1-2) SAPT vs 4 (4-4) HCL (p=0.8774). A1C significantly decreased from baseline to 2nd trimester, and was different between groups in the 3rd trimester (Table 1). There were no differences in time below, within, or above range between groups during the study (Table 1) or in gestational health outcomes (Table 2). In a pilot cohort of pregnant women with T1D, assisted HCL therapy did not increase risk to mothers or fetuses compared to SAPT despite lack of a pregnancy-specific algorithm in the HCL system.

Disclosure

E.Buschur: Consultant; Beta Bionics, Inc., Research Support; Dexcom, Inc., JDRF, Leona M. and Harry B. Helmsley Charitable Trust. E.R.Nease: None. S.Polsky: Other Relationship; diaTribe, Research Support; Dexcom, Inc., Medtronic, Hemsley Charitable Trust, JDRF, NIH - National Institutes of Health. J.K.Snell-bergeon: None. L.Pyle: None. R.Garcetti: None. K.M.Dungan: Board Member; Elsevier, Consultant; Eli Lilly and Company, Dexcom, Inc., Other Relationship; UpToDate, Research Support; Dexcom, Inc., Abbott, ViaCyte, Inc., Sanofi, Speaker's Bureau; Academy for Continued Healthcare Learning, Cardiometabolic Health Congress, Medscape, Integritas. S.F.Thung: None. A.Bartholomew: None. T.Summerfield: None. T.Bloks: None.

Funding

JDRF (2-SRA-2018-666-M-R); Colorado Clinical and Translational Sciences Institute; Children’s Diabetes Foundation; National Center for Research Resources (UL1RR02578)
1756-P: Extracellular Matrix Stiffness Mediates Insulin Secretion in Pancreatic Islets via Phosphofructokinase Activity
Authors: GARCIA CHELSEA , HOLCOMB KEIFER , FARNSWORTH NIKKI L.
Published: June 2023

Pancreatic islets are surrounded by extracellular matrix (ECM) which provides mechanical and biochemical cues to the β-cells that regulate survival and insulin secretion. Changes to ECM stiffness have been correlated to dysfunction in insulin secretion that is directly coupled to glucose metabolism; however, the mechanisms of mechanotransduction regulating insulin secretion have not been studied in the islet. We hypothesize that increasing matrix stiffness will increase islet glucose sensitivity by increasing phosphofructokinase (PFK) activity. To determine the effect of changes in ECM stiffness on islet function we encapsulated mouse islets in a reverse thermal gel (RTG) scaffold with increasing stiffness as determined by rheological analysis. Glucose-stimulated insulin secretion, PFK activity, and PFK expression was measured after 24 hours of culture. Proliferation was analyzed by immunohistochemistry of Ki-67. Reactive oxygen species (ROS) were measured via confocal microscopy of islets transfected with an adenoviral ROS sensor. We found that increasing RTG wt% yielded increasing stiffness at 40°C. Insulin secretion increased as the matrix stiffness increased in basal (2mM) and high glucose (20mM) conditions. Insulin secretion at 20mM glucose normalized to 2mM glucose (stimulation index) decreases with matrix stiffness indicating dysfunction to insulin secretion. PFK activity increased in islets encapsulated in stiffer RTGs. No changes in proliferation were observed in any samples compared to controls. Our results support a role for ECM stiffness in regulating insulin secretion via PFK activity. This work provides insight into how changes in ECM stiffness contribute to islet dysfunction, as in pancreatitis and T2D. Our results will inform future development of mechanically tuned microenvironments for differentiation of iPSCs into functional β-cells and provide novel targets to protect against islet dysfunction in T2D.

Disclosure

C.Garcia: None. K.Holcomb: None. N.L.Farnsworth: None.

Funding

American Diabetes Association (7-21-JDF-020 to N.L.F.); National Institutes of Health (1F31DK132926-01A1); JDRF (3-APF-2019-749-A-N, 1-FAC-2020-891-A-N)
1774-P: Verapamil Prevents Decline of IGF-1 in Subjects with T1D
Authors: XU GUANLAN , CHEN JUNQIN , LU BRIAN , QIAN WEIJUN , SHALEV ANATH
Published: June 2023

We have previously found that the calcium channel blocker and anti-hypertensive, verapamil, promotes functional beta cell mass and improves glucose homeostasis in diabetic mice and in humans with recent-onset type 1 diabetes (T1D). Surprisingly, we also discovered that verapamil reversed the T1D-associated elevation in T-follicular-helper cell markers in peripheral blood monocytes, suggesting that it also has immunomodulatory effects. Now, a global proteomics analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) of serum samples from T1D subjects at baseline and after 1 year of receiving verapamil or placebo identified insulin-like growth factor 1 (IGF-1) as one of the top proteins with significantly changed abundance over time. Specifically, IGF-1 abundance decreased during year 1 after diagnosis in subjects with T1D receiving placebo, which is consistent with the lower IGF-1 levels reported previously in individuals with T1D as compared to age- and sex-matched healthy subjects. Interestingly, this decline was blunted in subjects receiving verapamil. To further confirm the LC-MS/MS results, we also measured patient serum levels of IGF-1 by ELISA. The results were very much in alignment and again revealed significantly decreased IGF-1 levels in the placebo group a year after diagnosis, but no such effect in the verapamil treatment group, suggesting that verapamil prevents the decline of IGF-1 in subjects with T1D. Of note, IGF-1, which is primarily produced in liver, acts on many tissues, including pancreatic islets promoting pathways involved in cell growth, differentiation and glucose metabolism. In beta cells, IGF-1 has also been shown to control apoptosis. Thus, our results reveal yet another previously unappreciated pathway affected by verapamil that may contribute to the beneficial effects observed with this treatment in the context of T1D.

Disclosure

G.Xu: None. J.Chen: None. B.Lu: None. W.Qian: None. A.Shalev: Other Relationship; TIXiMED.

Funding

National Institutes of Health (R01DK078752, U01DK120379)
1776-P: Glucolipotoxic Conditions Affect Membrane Potential Wave Velocity Determined in Intact Islets Using Microelectrode Arrays
Authors: GRESCH ANNE K. , DEICKERT THOMAS , HÜWEL JAN D. , BRIGGS JENNIFER , BEECKS CHRISTIAN , BENNINGER RICHARD K. , DÜFER MARTINA
Published: June 2023

Pancreatic β-cells form highly connected networks within pancreatic islets. Each cell shows upon stimulation an oscillatory depolarization of the membrane potential and subsequent changes in [Ca2+]c. These signals propagate through the islet. The extent to which this coordinated islet cell activity is determined by a subset of β-cells is currently under discussion. In order to determine factors that influence this propagation, we have chosen a new method to identify wave propagation. We used high-resolution microelectrode arrays (CMOS-MEA) to measure voltage changes in the membrane potential of islet cells within the intact mouse islet. For network analysis, the individual electrode traces were compared with each other to gain insight into connectivity. The node degree of an islet (treated with 10 mM glucose) appears to be higher in the middle, but on average it was 8±2 degrees. Furthermore, algorithms were established to track the wave propagation from electrode to electrode, i.e. approximately from cell to cell. Treatment of isolated islets from C57BL6 mice with increasing glucose concentrations revealed that 15 mM glucose significantly reduced the wave duration compared to 6, 8 and 10 mM glucose (6 mM glucose: 10±4 s, n=32, vs. 15 mM glucose 7±2 s, n=26, p=0.0007). Additionally, the wave velocity was increased with rising glucose concentrations (6 mM glucose: 72±33 µm/s, n=20, vs. 8 mM glucose 118±69 µm/s, n=25, p=0.009). However, when the islets were pre-incubated with glucolipotoxic medium (25 mM glucose, 100 µM palmitate) for 72 h to mimic pre-diabetic conditions the wave velocity (waves induced by 8 mM glucose) was reduced (control: 87±40 µm/s, n=19, vs. GLT 64±32 µm/s, n=25, p=0.04). The results show that propagating waves of membrane depolarization are likely an important aspect for a functioning islet. The established technique proves to be suitable for tracking signal propagation and opens a wide field for further studies on islet network signaling.

Disclosure

A.K.Gresch: None. T.Deickert: None. J.D.Hüwel: None. J.Briggs: None. C.Beecks: None. R.K.Benninger: None. M.Düfer: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (R01DK102950R01, DK106412)
1779-P: Lipotoxicity-Induced Beta-Cell Small Extracellular Vesicles Promote Beta-Cell Functional Failure through Activation of the TGFß/Smad3 Pathway
Authors: ROY ABHISHEK , HOFF ALEXANDRA , HER TRACY K. , SANKARAN RAJAGOPALAN KAMALNATH , MALTAIS DANIELA D. , MATVEYENKO ALEKSEY , JAVEED NAUREEN
Published: June 2023

Lipotoxicity is a pathological process that occurs in several metabolic disorders including obesity-driven Type 2 diabetes (T2D). Chronically elevated circulating free fatty acids have been shown to exert detrimental effects on pancreatic islet β-cells through reduced glucose-stimulated insulin secretion (GSIS), alterations in β-cell transcriptional identity, and apoptosis. β-cell-derived small extracellular vesicles (sEV) have been shown to contribute to β-cell failure in T2D, however their role in lipotoxicity-mediated β-cell failure has yet to be elucidated. We hypothesize that lipotoxicity-induced β-cell functional failure is mediated in part by aberrant release of altered β-cell-derived sEV. To test this, MIN6 β-cells were exposed to free fatty acid, palmitate (PAL) (0.5 mM, 24h) and EV were isolated using differential ultracentrifugation to yield PAL EV (vs. control (CTL) EV). β-cell functional assessment using GSIS on mouse islets exposed to PAL EV (48h) resulted in significant suppression of GSIS (~80% decrease in stimulation index). RNA-Seq was conducted on islets exposed to PAL EV (vs. untreated (UT)). De novo motif analysis from this data identified enriched transcription factor binding motifs associated with several SMAD genes in TGFβ signaling. Gene Set Enrichment Analysis revealed significant enrichment of genes associated with TGFβ receptor binding suggesting its potential role in lipotoxic β-cell EV-mediated β-cell failure. To confirm, analysis of p-Smad3 expression was conducted by immunofluorescence staining on mouse islets exposed to PAL EV (48h; vs. UT). The results showed an ~60-fold increase in β-cell p-Smad3 expression (p<.05). Moreover, mouse islets were treated with a TGFβ receptor inhibitor in the presence of PAL EV (48h; vs. UT). GSIS results showed a significant improvement in β-cell function (p<.05) suggesting the role of the TGFβ/Smad3 pathway in lipotoxic β-cell sEV induced β-cell failure.

Disclosure

A.Roy: None. A.Hoff: None. T.K.Her: None. K.Sankaran rajagopalan: None. D.D.Maltais: None. A.Matveyenko: None. N.Javeed: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (DK129208-01); JDRF (2-SRA-2022-1272-S-B)
1780-P: Assessment of Toll-Like Receptor Expression and Signaling in Islets under Metabolic and Inflammatory Conditions
Authors: MATTKE JORDAN D. , NAZIRUDDIN BASHOO , LAWRENCE MICHAEL C.
Published: June 2023

Pancreatic beta cells undergoing metabolic and inflammatory stress express islet-derived cytokines or “isletokines” which can induce immune cell infiltration and cellular damage leading to loss of islet cell function. We and others have shown that toll-like receptor (TLR) 4 in islets can mediate metabolic and inflammatory response to DAMPs under diabetic conditions and in islet cell transplantation. Here we show that metabolic and inflammatory conditions result in induction and suppression of expression of multiple members of the TLR family in both MIN6 beta-cell line and mouse islets. Among upregulated TLRs, the endosomal nucleic acid-sensing TLR3 was most highly induced (up to 5 fold) in response to cytokine cocktail IL-1beta, TNF-alpha, and IFN-gamma in the presence of high glucose. Induction of TLR3 expression was blocked by PI3 kinase inhibitor wortmannin and unaffected by mTOR inhibitor rampamycin, calcineurin inhibitor FK506, and NF-κB inhibitor BMS345541. Immunoblot analyses confirmed that TLR3 but not TLR4 protein expression was wortmannin sensitive. These data indicate that TLR3 is upregulated by PI3 kinase signaling in islet beta cells in response to inflammatory conditions. As the endosomal TLR3 signaling is known to sense nucleic acids as well as influence glucose homeostasis, we propose that beta cells may sense DNA damage to induce isletokines to mediate local inflammatory responses during metabolic and inflammatory stress. Further investigations are warranted to decipher signaling pathways and molecular targets of TLR3 signaling to alleviate islet inflammation and improve beta-cell function.

Disclosure

J.D.Mattke: None. B.Naziruddin: None. M.C.Lawrence: None.
180-LB: Associations of Phthalate Metabolites with Glycemic Parameters and Fetal Growth in Type 1 Diabetes (T1D) Pregnancies
Authors: PRESTON EMMA V. , HANSBURY GEORGIA A. , O'CONNELL JULIANNE C. , WOLFS DANIELLE , DREYFUSS JONATHAN , WOLPOWITZ SHIRA , WANG ZIFAN , JAMES-TODD TAMARRA , ISGANAITIS ELVIRA M. , BROWN FLORENCE M.
Published: June 2023

Background: The risk of delivering a large for gestational age (LGA) infant is higher in pregnant individuals with T1D compared to individuals without diabetes, despite advances in managing T1D in pregnancy. Exposure to endocrine disrupting chemicals (e.g., phthalates) can affect both glucose homeostasis and fetal growth.

Objective: To estimate associations of urinary phthalate metabolite levels with continuous glucose monitoring (CGM) data in pregnancy and birthweight.

Methods: We analyzed urinary levels of 14 phthalate metabolites and derived trimester-specific CGM summary measures in 32 pregnant women with T1D. We estimated associations of log2-transformed urinary metabolite levels with continuous CGM outcomes and infant weight (GROW percentile) using linear regression, and estimated associations between metabolite levels and odds of LGA (GROW percentile ≥90) vs. AGA (<90) using logistic regression. Models were adjusted for maternal age and gestational week of study visit.

Results: Participants were primarily white (85%), nulliparous (53%), with a mean age of 33 years, and 50% had LGA infants. Concentrations of mono-benzyl phthalate (mBzP) and mono-carboxy octyl phthalate (mCNP), were associated with glycemia in the 3rd trimester. Specifically, each doubling of mBzP and mCNP concentration was associated with 8.5 mg/dL (95% CI: 1.4, 15.6) and 8.9 mg/dL (-1.3, 19.0) higher average glucose, -6.0% (-10.9, -1.0) and -8.9% (-15.1, -2.7) less time spent in range (63-140mg/dL), and 5.9% (0.7, 11.2) and 7.4% (0.3, 14.5) more time spent above range (>140 mg/dL). Phthalates were not associated with continuous GROW percentile.

Conclusion: Exposure to certain phthalates in later pregnancy may affect glycemic control in pregnant individuals with T1D. However, effects of phthalate exposure on birthweight are unclear. Future studies may disentangle the extent to which phthalate exposure and its effects on glycemic control may impact fetal growth.

Disclosure

E. V. Preston: None. F. M. Brown: Other Relationship; Dexcom, Inc. G. A. Hansbury: None. J. C. O'connell: None. D. Wolfs: None. J. Dreyfuss: None. S. Wolpowitz: None. Z. Wang: None. T. James-todd: None. E. M. Isganaitis: None.

Funding

National Institute of Environmental Health Sciences; National Institute of Diabetes and Digestive and Kidney Diseases
180-OR: Baseline Predictors of Remission of Type 2 Diabetes in Response to Short-Term Insulin-Based Therapy—The Pivotal Role of Beta-Cell Function
Authors: RETNAKARAN RAVI , PU JIAJIE , EMERY ALEXANDRA , KRAMER CAROLINE K. , ZINMAN BERNARD
Published: June 2023

Short-term treatment with insulin can induce remission of type 2 diabetes (T2DM) in some patients but not others. We thus sought to identify baseline predictors of diabetes remission in response to short-term insulin-based therapy. In this study, adult patients with T2DM of <7 years duration were randomized to 8-weeks treatment with (i) insulin glargine, (ii) glargine + thrice-daily lispro, or (iii) glargine + twice-daily exenatide, followed by 12-weeks washout that enabled assessment of remission (defined as A1c<6.5% after ≥3 months without glucose-lowering therapy). At baseline, 8-weeks, and washout, beta-cell function was assessed with 4 measures: Insulin Secretion-Sensitivity Index-2 (ISSI-2), insulinogenic index/HOMA-IR, ΔCpeptide0-120/Δglucose0-120×Matsuda, and ΔISR0-120/Δgluc0-120×Matsuda. Diabetes remission was achieved in 31 of 90 participants (34.4%). Compared to their peers, those who went on to remission had lower A1c (p<0.001) and better beta-cell function at baseline (all 4 measures p≤0.01). The non-remission and remission groups did not otherwise differ in baseline insulin sensitivity/resistance (Matsuda, HOMA-IR), BMI, duration of diabetes, pre-trial diabetes medications, or allocated insulin-based therapy during the trial. On logistic regression analyses, each baseline measure of beta-cell function emerged as a significant predictor of remission (log ISSI-2: adjusted OR 4.41 (95%CI: 1.71-11.34); log insulinogenic index/HOMA-IR: 2.21 (1.26-3.89); log ΔCpeptide0-120/Δglucose0-120×Matsuda: 1.62 (1.00-2.64); log ΔISR0-120/Δgluc0-120×Matsuda: 1.87 (1.09-3.23). Similarly, higher baseline ISSI-2 tertile predicted longer time to glycemic relapse after cessation of the insulin-based therapy (log-rank P=0.029). In conclusion, beta-cell function is the dominant baseline determinant of the likelihood of achieving remission of diabetes in response to short-term insulin-based therapy.

Disclosure

R.Retnakaran: Other Relationship; Sanofi, Eli Lilly and Company, Research Support; Boehringer Ingelheim/Mount Sinai Hospital, Novo Nordisk. J.Pu: None. A.Emery: None. C.K.Kramer: Research Support; Boehringer Ingelheim Inc. B.Zinman: Consultant; Abbott Diabetes, Eli Lilly and Company, Novo Nordisk A/S, Novo Nordisk Canada Inc., Boehringer Ingelheim Inc., Merck & Co., Inc.

Funding

Canadian Institutes of Health Research (MOP136938)
181-OR: Pancreatic Gene Therapy Durably Improves Glycemia and Delays Disease Progression in a Murine Model of Type 2 Diabetes
Authors: FITZPATRICK ALICE L. , COZZI EMILY , WAINER JACOB , REESE REBECCA , KHANNA NIDHI , CAPLAN JAY , RAJAGOPALAN HARITH , WEST JASON A.
Published: June 2023

Beta-cell (β-cell) failure is a root cause of the progression of type 2 diabetes (T2D). GLP-1 receptor agonists (GLP1RA) have proven clinical efficacy and improve β-cell function. However, systemic side effects limit patient adherence and long-term efficacy. Localized pancreatic GLP1RA production may engender durable islet cell benefit while diminishing side effects. We developed a novel adeno-associated virus (AAV) gene therapy delivery platform to enable local and durable production of therapeutic proteins by the pancreas. We assessed to what degree a β-cell restricted GLP1RA gene therapy impacts disease progression in the db/db mouse compared to semaglutide (Sema).

To establish a baseline for chronically administered systemic GLP1RA, Sema (10 nmol/kg) or vehicle were given daily to 5-week-old mice for 10 weeks (n=8/group). Sema treated mice had a 64% decrease in fasting blood glucose (FBG; p<0.0001) and a 3.5-fold increase in fasting insulin (FI; p=0.01) across the time course. Glucose tolerance was significantly improved during IPGTT at weeks 3, 6, and 9 (p<0.001).

To determine the impact of a single dose GLP1RA gene therapy, 1) β-cell restricted AAV-GLP1RA, 2) AAV control, or 3) vehicle were given to 5-week-old mice (n=8/group). Remarkably, AAV-GLP1RA treatment decreased FBG levels by 70% (p<0.0001) and elevated FI by 1.9-fold (p<0.01) compared to controls for 10 weeks. Glucose tolerance was improved during IPGTT at weeks 3, 5, 7, and 9 (p<0.01). AAV-GLP1RA protein was undetectable in serum and restricted to the islet, suggesting improved glycemic control was due to intra-islet GLP1RA production and signaling.

These results demonstrate that a single dose β-cell restricted GLP1RA gene therapy can comparably improve glycemia and delay disease progression in a murine model of T2D when compared to chronic systemic GLP1RA administration. These data provide proof of concept that a gene therapy has the potential to durably treat metabolic diseases, including T2D.

Disclosure

A.L.Fitzpatrick: Employee; Fractyl Health, Inc. E.Cozzi: Employee; Fractyl Health, Inc. J.Wainer: Employee; Fractyl Health, Inc. R.Reese: Employee; Fractyl Health, Inc. N.Khanna: Employee; Fractyl Health, Inc. J.Caplan: Employee; Fractyl Health, Inc. H.Rajagopalan: Employee; Fractyl Health, Inc. J.A.West: Employee; Fractyl Health, Inc.

Funding

Fractyl Health, Inc.