bluuebunny/crossref_metadata_2025_split huggingface dataset:
1996-LB: Relative Contributions of Glycosylated Hemoglobin and Glycemic Variability to the Development of Microvascular Outcomes from DCCT Study
Authors: ZHANG JIE , SONG ZHENQIANG , LI MINGZHEN , ZHU HONG , GAO SHAN , LIU YUFU 
Published: June 2024

Introduction and Object: As two key profiles for blood glucose control and complications, the relative contributions of glycosylated hemoglobin (HbA1c) and glycemic variability (GV) to microvascular outcomes are explored in type 1 diabetes mellitus in Diabetes Control and Complications Trial (DCCT) study.
Methods: Measures of GVs included the SD, mean amplitude of glycemic excursions (MAGE), coefficient of variation (CV) within and between quarterly 7-point glucose profiles and HbA1c.The generalized boosted models were used to analysis the relative effects of HbA1c and GV.
Results: Mean HbA1c accounted for 57%-62% /55%-57%for the development/progression of retinopathy, and GV accounted for 38%-43%/43%-57%, even similar in subgroup analysis including gender, age, duration, triglyceride and changes of nephropathy. HbA1c accounted for 46.5% for the occurrence of microalbuminuria, as for GV, 53.5% respectively. MAGE and SD accounted more in patients with duration of more than 10years. GV contributed more in subgroup with serum triglyceride≤55mg/dL and non-deteriorated retinopathy (58%-61%, 59%-67% respectively) in microalbuminuria.
Conclusion: Our finding showed that GV contributed equally as HbA1c, or more for microvascular outcomes in type 1 diabetes, especially for microalbuminuria. Refined and comprehensive blood glucose management would be needed for health care provides.

Disclosure
J. Zhang: None. Z. Song: None. M. Li: None. H. Zhu: None. S. Gao: None. Y. Liu: None.
20-PUB: Intensification of Human Insulin Therapy with SGLT2 Inhibitors in Patients with Type 2 Diabetes Mellitus
Authors: DZIDA GRZEGORZ J. , GAJDA PIOTR , MASIEREK MALGORZATA 
Published: June 2024

Treatment intensification with SGLT2 inhibitors added to insulin therapy is recommended in many T2DM guidelines as well as registered indications. The aim of the study was the assessment of the efficacy and safety of SGLT2 inhibitors with human insulin combinations in comparison with the general treatment options in T2DM patients onwith human insulins therapy. The data were collected in non-interventional multicenter observational open label study recruiting 4384 T2DM patients treated with human insulin (Gensulin®) injected with the same device (Gensupen®2 ). Combinations including SGLT2 inhibitors was observed in 1079 (24,6%). Triple combination of human insulin+metformin+SGLT2 inhibitors was the most commonly used (632 patients, 14,4%). SGLT2 inhibitors treatment intensification was found in 40,1% of patients with diagnosed HF (those with HFrEF in 59,65%), history of MI 40,4% and CKD in 30,25%. Patients age, gender, BMI nor history of stroke did not affect the frequency of SGLT2 inhibitors use. The decrease of A1c in 6 months observation period in SGLT2 group was 0,8% while 0,7% in the general treatment group. A1c after 6 months was similar in both groups (7,7% and 7,8%). The percentage of patients achieving target A1c less than 7% was also similar ( 5,6% and 21,3% after 6 months vs. 5,8% and 18,9% respectively). Median insulin daily dose didn’t differ between groups (35,2U vs 34U ) as well as insulin dose per kg (0,4U/kg vs.0,4U/kg) at the study end. The confirmed hypoglycaemia rate was very low and did not differ between groups as well as percentage of patients not reporting hypogylaemia in 6 month period ( 82,7% vs. 84%). The intensification of human insulin treatment in T2DM patients with SGLT2 inhibitors did not affect significantly glucose control efficacy and safety. HF, history of MI and CKD were the strongest indications for SGLT2 inhibitors choice. The optimalization of insulin dose seems to be a crucial intervention needed to achieve appropriate glucose control.

Disclosure
G.J. Dzida: None. P. Gajda: None. M. Masierek: Employee; BIOTON.
2000-LB: Multiome Profiling of Human Islets Nominates Beta-Cell–Specific Cytokine Response Profiles Underlying Type 1 Diabetes
Authors: ROBERTSON CASSIE , WANG XIAOOU , ORCHARD PETER , MENG JERRY , DOLGACHEV VLADISLAV , MANICKAM NANDINI , NARISU NARISU , ERDOS MICHAEL , COLLINS FRANCIS S. , CHEN SHUIBING , PARKER STEPHEN C. 
Published: June 2024

Inflammation in the islet may be an early trigger of beta cell stress which can lead to islet autoimmunity and type 1 diabetes (T1D). We exposed human islets (N=8 donors) to proinflammatory cytokines (IL-1-beta and TNF-alpha) and used single nucleus multiomic sequencing to jointly profile RNA expression and chromatin accessibility (10X Genomics Multiome). We pooled cytokine-treated and untreated islets for single nucleus experiments using a randomized block design, which reduced sample-to-sample variability and eliminated batch confounding of treatment effects. We included 37,170 high quality nuclei in downstream analysis and identified transcriptional response to cytokines in three major islet cell types (887 genes in alpha cells, 2,998 genes in beta cells, 547 genes in ductal cells), including upregulation of MHC class I in all three cell types. Beta cells showed significant downregulation in marker gene expression (INS fold change = 0.55, p = 3 x 10-5), but marker gene expression did not change in alpha (GCG fold change in alpha cells = 0.68, p = 0.07) or ductal cells (KRT19 fold change in ductal cells = 0.82, p = 0.60). Pathway analyses indicated upregulation of proinflammatory pathways in all three cell types and downregulation of a hallmark beta cell pathway in beta cells only (p = 0.005). Finally, pathway analysis of cell type-specific gene expression changes, using residualized treatment effect sizes per cell type, indicated enhanced upregulation of the TGF-beta signaling pathway in beta cells as compared to alpha or ductal cells. In conclusion, using single nucleus profiling with a randomized study design, we found beta cell-specific responses to proinflammatory cytokines which may contribute to beta cell demise underlying type 1 diabetes. Analyses of cytokine effects on chromatin accessibility to nominate upstream transcriptional regulators are underway.

Disclosure
C. Robertson: None. X. Wang: None. P. Orchard: None. J. Meng: None. V. Dolgachev: None. N. Manickam: Employee; 10x Genomics. N. Narisu: None. M. Erdos: None. F.S. Collins: None. S. Chen: None. S.C. Parker: Research Support; Pfizer Inc.

Funding
NIH NIDDK (DK127777); NIH NIDDK (DK007245)
2031-LB: Serotonin Is a Novel Player in Selective Hepatic Insulin Resistance
Authors: EUN NAM JUNG , HWANG INSEON , CHOI WON-IL , LEE MINJU , CHOI WONGUN , CHOI WONSUK , MOON YOUNG-AH , KIM HAIL 
Published: June 2024

Insulin is known to induce lipogenesis and inhibit gluconeogenesis in the liver. Therefore, hepatic lipogenesis is supposed to be downregulated in insulin resistance, but, in reality, hepatic lipogenesis is paradoxically upregulated. Recently, Serotonin (5-HT) has been implicated in both hepatic lipogenesis and gluconeogenesis. However, the molecular mechanism and the potential crosstalk between 5-HT and insulin signaling pathways have not been explored yet. Here, we investigated the multifaceted roles of 5-HT in the liver which may provide new insights into the complex regulatory networks governing hepatic glucose and lipid homeostasis.
AML-12 cells were incubated with 5-HT in the presence or absence of various inhibitors. Signaling pathways were explored in liver specific Htr2a (HTR2ALKO), Htr2b (HTR2BLKO), Insr (LIRKO) and both Htr2a and Insr (HTR2A/IRLKO) knockout mice after injection of 5-HT or insulin via portal vein.
In AML-12 cells, 5-HT activates AKT phosphorylation through PI3K and intracellular calcium flux resulting in the activation of SREBP-1c. Direct injection of 5-HT through portal vein induced activation of AKT and SREBP-1c, which were diminished specifically in HTR2ALKO mice. When the hepatic insulin signaling was inhibited by LIRKO or S961, 5-HT induced AKT phosphorylation but insulin could not. According to HFD fed condition, weight and size of liver and hepatic steatosis are decreased in HTR2A/IRLKO mice, comparing LIRKO mice. In terms of gluconeogenesis, 5-HT phosphorylates CREB via HTR2B which was mediated by PKG, not by PKA and 5-HT injection could not induce CREB activation in HTR2BLKO mice.
These results suggest that during insulin resistance 5-HT can increase both lipogenesis and gluconeogenesis in the liver via HTR2A-PI3K-AKT-SREBP1c signaling and HTR2B-PKG-CREB signaling, respectively. Therefore, the selective hepatic insulin resistance could be attributed to the 5-HT induced activation of lipogenesis and gluconeogenesis, independently from insulin and glucagon.

Disclosure
J. Nam: None. I. Hwang: None. W. Choi: None. W. Choi: None. W. Choi: None. Y. Moon: None. H. Kim: None.

Funding
NRF-2021R1A2C3005108; NRF-2020M3A9E4038695
2036-LB: Identification of a Hepatocyte Subpopulation That Simultaneously Expresses Gluconeogenic and Lipogenic Genes in the Fed State
Authors: OKADA JUNICHI , LANDGRAF AUSTIN , XIAOLI ALUS , QIU YUNPING , LIU LI , SCHUSTER VICTOR L. , KURLAND IRWIN J. , YANG FAJUN , ELISCOVICH CAROLINA , SHINODA KOSAKU , PESSIN JEFFREY 
Published: June 2024

Introduction & Objective: Pathway-selective insulin resistance in the liver is a concept where gluconeogenesis is selectively insulin resistant compared to de novo lipogenesis. Thus, treatment with large doses of insulin to control gluconeogenesis might lead to excessive triglyceride synthesis. To understand the discrepancy in the pathogenesis between the two pathways, we examined the zonation of hepatocyte specificity of gluconeogenic and lipogenic gene expression under normal physiological conditions in mice.
Methods: Using a combination of single cell RNA sequencing (scRNA-seq) and single molecule Fluorescence In Situ Hybridization (smFISH), we examined the normal regulation of gluconeogenic genes (such as Pck1) and lipogenic genes (such as Fasn) under various feeding/fasting conditions. Mass spectrometry with stable isotope using isolated pericentral and periportal hepatocytes confirmed gene expression results. PrimeFlow RNA Assay was performed to further understand the relationship between these genes.
Results: scRNA-seq showed a subset of periportal hepatocytes in the fed state that expressed both Pck1 and Fasn. smFISH not only confirmed the scRNA-seq data but also showed a subset of periportal hepatocytes that co-transcribed Pck1 and Fasn in the fed state. Mass spectrometry with stable isotope confirmed basal gluconeogenesis in the periportal hepatocytes in the fed state, suggesting that the subset of hepatocytes was undergoing gluconeogenesis in the fed state. PrimeFlow RNA assay followed by FACS enabled us to isolate these hepatocytes for further analysis.
Conclusion: We have discovered a new subset of hepatocytes that express both lipogenic and glucogenic genes under normal physiological conditions. These hepatocytes are by default pathway-selectively insulin resistance. Analyzing and characterizing as well as possibly targeting these hepatocytes might help us understand the pathogenesis of insulin resistance.

Disclosure
J. Okada: None. A. Landgraf: None. A. Xiaoli: None. Y. Qiu: None. L. Liu: None. I.J. Kurland: None. F. Yang: None. C. Eliscovich: None. K. Shinoda: None. J. Pessin: None.
2046-LB: Selective, Acute Stimulation of Sympathetic Activity in Brown Adipose Tissue Improves Glycemic and Lipid Profiles in Mice
Authors: NERI DANIELE , FOHN ALEXIS , BOZEC DOMINIQUE , HEINE MARKUS , WORTHMANN ANNA , CHEN XINHONG , HEEREN JOERG , ZELTSER LORI M. 
Published: June 2024

Brown adipose tissue (BAT) activity is linked to improved cardiometabolic health, and acute stimulation of BAT has been shown to reduce glycemia and triglyceridemia, which is protective against metabolic diseases such as type 2 diabetes. However, most BAT-stimulating protocols involve deliberate cold exposure or sympathomimetic drugs. Although effective, these methods can increase cardiovascular risk by increasing general sympathetic tone. Our lab has developed a model to selectively activate the sympathetic nerves controlling BAT activity in mice, and we are now dissecting the contribution of BAT activity to systemic metabolism without affecting other peripheral organs. We have used a combination of genetically engineered mouse lines, neurotrophic viruses, chemogenetics, metabolic recordings, and next generation sequencing to carefully study how the sympathetic nervous system controls different aspects of BAT activity. Our results show that the selective acute activation of BAT-projecting sympathetic nerves improves glucose tolerance in mice. This acute stimulation also increases total energy expenditure and lowers the respiratory exchange ratio, which is consistent with increased BAT activity. Finally, we show that the transcriptomic profile of BAT in these mice is in line with expected changes associated with increased BAT activity. BAT represents one of the most promising targets for new therapeutics to restore euglycemia. Our novel approach to selective sympathetic regulation pinpoints the undisputed role of BAT to whole-body glucose and energy management in mice. This targeted activation holds promises for novel therapies against type 2 diabetes while circumventing potential cardiovascular risks associated with traditional BAT stimulation methods.

Disclosure
D. Neri: None. A. Fohn: None. M. Heine: None. A. Worthmann: None. X. Chen: None. J. Heeren: None. L.M. Zeltser: None.

Funding
R01DK125094; P01AG032959
215-OR: Polygenic Scores for Metabolic Diseases Are Associated with Gene Expression Networks across Tissues
Authors: BRYAN CHRISTOPHER , SMITH KIRK , MANNING ALISA , MERCADER JOSEP M. , UDLER MIRIAM 
Published: June 2024

In type 2 diabetes (T2D), coronary artery disease (CAD) and obesity, disease risk across thousands of genetic variants can be captured with global extended polygenic scores (gePS), yet knowledge of underlying disease pathways remains limited. Expression Quantitative Trait Scores (eQTS) connecting gePS to gene expression could help prioritize disease-causing genes and pathways in specific tissues.
We generated gePS for T2D, CAD, and body mass index (BMI) in the Genotype-Tissue Expression (GTEx) database of 838 individuals and 49 tissues. Associations between gePS and phenotypes were confirmed in a hospital-based biobank. eQTS were calculated using a linear model of association between gePS and tissue transcript expression level, adjusting for age, sex, and 5 PCs. Gene set enrichment was performed using Enrichr with Gene Ontology pathway sets and Fisher’s Exact Test (Q &lt; 0.05). Each gePS was strongly associated with &gt;3,000 strong transcript expression levels (eQTS) across multiple tissues at Q&lt;0.05. T2D and BMI eQTS in multiple tissues were each significantly enriched for pathways related to mitochondrial function (Q&lt;0.01), with the most significant T2D signal in the heart atrial appendage and BMI signal in subcutaneous (SC) and visceral adipose. 3,960 eQTS were shared between T2D and BMI in the heart left ventricle (LV), and 68 were shared in SC adipose (all Q&lt;0.05). Shared pathways in the heart were enriched for processes related to cellular respiration and heart contraction (Q&lt;10-8). For CAD, cholesterol metabolism pathways were enriched in the coronary artery and visceral adipose (Q&lt;10-2). In summary, T2D and BMI eQTS share mitochondrial and metabolic transcription mechanisms across several tissues, including SC adipose. Cholesterol metabolism was enriched in multiple tissues in CAD eQTS and also observed in T2D and BMI eQTS in visceral adipose. This study provides novel insight into the shared genetic and transcriptomic architecture of T2D, CAD, and obesity.

Disclosure
C. Bryan: None. K. Smith: None. A. Manning: None. J.M. Mercader: None. M. Udler: Other Relationship; Up-To-Date.

Funding
Doris Duke Clinical Scientist Development Award (2022063)
218-OR: Disrupted Circadian Glucocorticoid Rhythms and High-Fat Diet Drive Obesity and Insulin Resistance through Distinct Mechanisms
Authors: TERUEL MARY 
Published: June 2024

Disrupted glucocorticoid (GC) rhythms, a hallmark of chronic stress and sleep dysregulation, are increasingly recognized as potential contributors to obesity and insulin resistance [1-3]. However, their impact compared to dietary factors remains unclear. Here we investigated the combined effects of high-fat diet (HFD) and disrupted GC rhythms on fat accumulation, metabolism, and lean mass in mice. Strikingly, we found that disrupted GC rhythms alone led to a doubling of adipose tissue mass within three weeks, comparable to HFD-fed mice, but without increased lipids in the liver or in circulating lipid and glucose levels. Mice with disrupted GC rhythms achieved a unique metabolic state of insulin resistance but surprisingly improved glucose tolerance. Mechanistically, we found this unique state was driven by persistent hyperinsulinemia to levels 10-fold higher than induced by HFD, despite persistently low glucose levels. The increase in insulin was driven by a rapid expansion of pancreatic beta cell mass and a shift towards cAMP-mediated insulin secretion. EchoMRI body composition measurements revealed a shift from lean-to-fat mass in GC-disrupted mice, resembling sarcopenic obesity. Our results demonstrate that disrupted GC rhythms can be just as potent as dietary excess in promoting fat mass gain. Furthermore, we found that disruption of GC rhythms and HFD increase fat mass through distinct and separable mechanisms, suggesting ways for effective personalized anti-obesity strategies that go beyond calorie restriction.
References:1. Bahrami-Nejad et al (2018). Cell Metabolism 27, 854-868.e8, 2. Luijten et al (2019). Cell Reports 27, 1686-1698. 3. Tholen et al (2022). Cell Reports 39, 111018.

Disclosure
M. Teruel: None.

Funding
National Institutes of Health (1-R01-DK131432)
225-OR: Sequential Islet Transplants for Type 1 Diabetes Do Not Cumulatively Increase Hepatic Portal Venous Pressure
Authors: CARR ALICE L.J. , SHAH RAHI , MARFIL-GARZA BRAULIO A. , LAM ANNA , ANDERSON BLAIRE L. , O'GORMAN DOUG , KIN TATSUYA , BIGAM DAVID , JAMES SHAPIRO A.M. , SENIOR PETER A. 
Published: June 2024

The future of islet transplantation (ITx) is rapidly moving towards stem-cell based therapies, which have led to short term insulin independence when delivered into the hepatic portal vein. It is likely that multiple infusions will be needed to maintain insulin independence long term. We examined the effects of sequential ITx on portal pressures and if features of the transplanted preparation influence acute changes in portal pressure. We include data from adults undergoing intraportal ITx at the University of Alberta Hospital between May 1999 to April 2023. We assessed, by pairwise comparison, acute changes in portal pressure at each ITx infusion and used linear mixed-effects models to: 1) compare sequential pre-infusion portal pressure with reference to pre-first ITx infusion and 2) assess the relationship between change in portal pressure and packed cell volume (PCV), purity and islet equivalents per kilogram (IEQ/kg). We include 299 adults (44% M) who received up to 5 (median of 2 [IQR 2-3]) ITx infusions and a median total 15,220 IEQ/kg [IQR 11,324, 19,700]. The median purity across all ITx infusions was 55% [IQR 45, 70] with a median PCV of 3ml [IQR 2.5, 4]. The total time to final ITx was a median 11m [IQR 2-50]. Portal pressure acutely increased after each ITX infusion (p&lt;0.001) by a median 2.0 mmHg [IQR 1.0-4.0]. This acute rise was not sustained however, with sequential pre-infusion portal pressure returning to similar levels (p&gt;0.1) as that pre-first ITx infusion (median 11.0 mmHg [IQR 8.0-14.0]). PCV had the strongest correlation with post-infusion change in portal pressure, with a 1 mmHg increase in portal pressure per 1 ml of PCV (Slope: 1.09 [95% CI0.93,1.26]; p&lt;0.001). Sequential ITx do not increase portal pressure or amplify changes during supplementary infusions. Higher PCV may exacerbate acute rises in portal pressures. Insights from deceased-donor intraportal ITx can inform safe implementation of future stem-cell based therapies in people living with diabetes.

Disclosure
A.L.J. Carr: None. R. Shah: None. B.A. Marfil-Garza: None. A. Lam: None. B.L. Anderson: None. D. O'Gorman: None. T. Kin: None. D. Bigam: None. A. Shapiro: Consultant; Vertex Pharmaceuticals Incorporated, Betalin Therapeutics, Hemostemix Inc, ViaCyte, Inc., Aspect Biosystems. P.A. Senior: Consultant; Abbott, Bayer Inc., Dexcom, Inc., Eli Lilly and Company, GlaxoSmithKline plc, Insulet Corporation, Novo Nordisk Canada Inc., Sanofi, Vertex Pharmaceuticals Incorporated. Speaker's Bureau; Abbott, Dexcom, Inc., Insulet Corporation, Novo Nordisk Canada Inc. Research Support; Novo Nordisk Canada Inc.
273-OR: Initial eQTL Discovery in Individual Pancreatic Cell Types from the Human Pancreas Analysis Program
Authors: WEIDEKAMP MARY A. , ELGAMAL RUTH , LORENZ KIMBERLY , KAESTNER KLAUS H. , GAULTON KYLE J. , GRANT STRUAN F. , VOIGHT BENJAMIN F. 
Published: June 2024

The pancreas is composed of multiple, distinct cell types with etiologically relevant functions for diabetes. Pancreatic studies have collected data largely from bulk tissue, limiting the elucidation of how each pancreatic cell type contributes to disease. Single-cell technologies coupled to tissue collection efforts have expanded for the pancreas, facilitating novel characterization of genetics, expression, and regulation of these cell types at a population scale and across disease states. A leading effort generating this data, the Human Pancreas Analysis Program, aims to comprehensively study the human pancreas in diabetes and integrate data with the clinical history from de-identified donors. Data from this program provides an opportunity to investigate how each of the pancreas’ cell types uniquely contributes to metabolic traits. Here, we report an initial discovery effort to identify genetic variation associated with gene transcript abundance - i.e., expression quantitative trait loci (eQTL) - using 394,151 cells collected from single-cell RNA sequencing of 80 pancreatic donors. Clustering analysis of this data identified pancreatic islets, pancreatic exocrine cells and immune cell types. We initially focused on populations of alpha, beta and delta cells, which had sufficient cell numbers across donors. Using standard analysis pipelines, we observed 1,696, 1,744 and 946 significant eGenes in alpha, beta and delta cells (permutation p-value &lt;0.05 and q-value &lt;0.05), respectively. Formal colocalization with signals identified from a recent type 2 diabetes GWAS (Vujkovic et al, 2020) and our eQTL data identified 19 SNP-to-gene connections, all of which were unique to one of our identified cell types. One eGene, NCK1, has been previously investigated in beta cells, but our colocalization has implicated it in an alpha cell context. Future colocalizations and meta-analyses should improve nomination of putative effector transcripts underlying pancreatic disease.

Disclosure
M.A. Weidekamp: None. R. Elgamal: None. K. Lorenz: None. K.H. Kaestner: None. K.J. Gaulton: Consultant; Pfizer Inc. Stock/Shareholder; Neurocrine. S.F. Grant: None. B.F. Voight: None.

Funding
UM1 DK126194; T2D FGPU01 DK123594; HPAP-T2DU01 DK112217; HPAP-T1D