bluuebunny/crossref_metadata_2025_split huggingface dataset:
2087-LB: Novel Human Islet Spheroid Models of Type 1 Diabetes
Authors: TITLE ALEXANDRA C. , RUFER CHANTAL , JAWUREK SAYRO , FORSCHLER FELIX , KODIYAN BEN , YESILDAG BURCAK 
Published: June 2024

Recently, it has emerged that β-cells co-orchestrate their own demise by increasing their visibility to the immune system, stressing the importance of studying both the immune and the β-cell components of T1D simultaneously. To enable accurate, high-throughput and longer-term studies of these mechanisms in vitro, we established three islet-immune injury models by culturing spheroids derived from primary human islets with proinflammatory cytokines, activated peripheral blood mononuclear cells or HLA-A2-restricted preproinsulin-specific cytotoxic T lymphocytes. We developed a wide variety of biochemical and 3D microscopy based-methods to assess dynamic and static insulin release, insulin production, β-cell survival, cytokine secretion and T cell infiltration, at a single islet resolution. In all models, the decline in β-cell health manifested as increased basal and decreased glucose-stimulated insulin release (GSIS), and decreased intracellular insulin content. Additional hallmarks of T1D progression such as loss of the first-phase insulin response (FFIR), increased proinsulin-to-insulin ratios, HLA-class I expression, and inflammatory cytokine release were also observed. Using these models, we evaluated the effects of multiple intervention strategies including a glucagon-like peptide 1 (GLP-1) receptor agonists and HLA Class I inhibition, on human islets under T1D-relevant stress. We were able to successfully prevent the loss of GSIS under cytokine- and immune-mediated stress, notably by increasing β-cell survival, preserving the FFIR, decreasing immune cell infiltration and cytokine secretion. Our results support the anti-inflammatory effect of GLP-1R agonists and importance of HLA Class I modulation for protecting β-cells from the immune-mediated attack that leads to T1D. The described biomimetic islet-immune assays represent ideal in vitro models for long-term, high-throughput and precise study of early-stage T1D immunopathology and β-cell dysfunction.

Disclosure
A.C. Title: Other Relationship; Biomea Fusion, Inc., Boehringer-Ingelheim, Novo Nordisk, Merck & Co., Inc. C. Rufer: Other Relationship; Boehringer-Ingelheim, Novo Nordisk, Merck & Co., Inc., Biomea Fusion, Inc. S. Jawurek: Other Relationship; Boehringer-Ingelheim, Merck & Co., Inc., Biomea Fusion, Inc., Novo Nordisk. F. Forschler: Other Relationship; Biomea Fusion, Inc., Boehringer-Ingelheim, Merck & Co., Inc., Novo Nordisk. B. Kodiyan: Other Relationship; Biomea Fusion, Inc., Novo Nordisk, Boehringer-Ingelheim, Merck & Co., Inc. B. Yesildag: Other Relationship; Novo Nordisk, Boehringer-Ingelheim, Biomea Fusion, Inc., Merck & Co., Inc.
2092-LB: Receptor-Mediated G12 Signaling Strongly Promotes Insulin Secretion from Pancreatic Beta Cells
Authors: PITTALA SRINIVAS , HASPULA DHANUSH , LIU LIU , CUI YINGHONG , WESS JÜRGEN 
Published: June 2024

Type 2 diabetes (T2D) has emerged as a major threat to human health in most parts of the world. The inability of pancreatic β-cells to release enough insulin is key pathological sign. Glucose-induced insulin secretion is regulated by G protein-coupled receptors (GPCRs) located on the cell surface of pancreatic β-cells. Because of the lack of GPCRs that can selectively interact with G proteins of the G12 family (G12/G13), it remains unknown whether G12 signaling has a positive impact on β-cell function. To address this question, we generated a new mouse line that expressed a CNO (clozapine-N-oxide)-sensitive, G12-coupled designer GPCR (G12 DREADD; DREADD = designer receptor exclusively activated by a designer drug) selectively in β-cells (beta-G12D mice). CNO is a small synthetic molecule that can selectively activate the G12 DREADD but is otherwise pharmacologically inert. In addition, we generated a mouse model lacking the α-subunits of G12 and G13 selectively in β-cells (beta-G12/13 KO mice). We then subjected these mutant mice, together with their control littermates, to a series of metabolic tests. We found that selective activation of β-cell G12 signaling strongly enhanced glucose-stimulated insulin secretion, leading to improved glucose tolerance in both lean and obese mice. In obese mice, activation of β-cell G12 signaling counteracted the deficits in glucose homeostasis caused by increased adiposity. The G12D-mediated simulation of insulin release was virtually abolished in beta-G12/13 KO mice. Mechanistic studies with perifused pancreatic islets demonstrated that the increase in insulin release triggered by stimulation of β-cell G12 signaling required the activation of ROCK1, leading to structural changes in the actin cytoskeleton that promoted insulin secretion. Our data strongly suggest that strategies aimed at stimulating β-cell G12 signaling could become clinically useful for the treatment of T2D and related metabolic disorders.

Disclosure
S. Pittala: None. D. Haspula: None. L. Liu: None. Y. Cui: None. J. Wess: None.

Funding
NIH Intramural Research Program
21-PUB: Meaningful Improvements in Treatment Satisfaction with Gla-300 in Insulin-Naïve PwT2D—A Post Hoc Analysis of Real-World ATOS PRO Study
Authors: HARRIS STEWART B. , KHAN NIAZ , TIROSH AMIR , KESAVADEV JOTHYDEV , VARGAS-URICOECHEA HERNANDO , ROBOREL DE CLIMENS AUDE , BAGHOUS HOUSSEM , SNOEK FRANK J. , MABUNAY MARIA AILEEN N. , GRULOVIC NATASA , CORP DIT GENTI VALERIE , MSIHID JEROME , GALSTYAN GAGIK R. 
Published: June 2024

Introduction: Initiating Gla-300 has shown to improve treatment satisfaction in PwT2D in RCT and real-world studies. ATOS, a 12-month prospective observational global study showed that initiating Gla-300 in insulin-naïve PwT2D resulted in improved glycemic control and low hypoglycemia rates.
Methods: This post-hoc analysis evaluated meaningful improvements in Diabetes Treatment Satisfaction Questionnaire status (DTSQs) scores with glycemic parameters. Data was collected at baseline, months 3, 6 and 12, and based on distribution-based method (0.5 SD of baseline DTSQs score) for meaningfulness, percentage of responders was determined.
Results: Total, 3801 participants completed the questionnaire. At baseline, mean ± SD age was 57.7 ± 10.5 years, duration of diabetes was 10.2 ± 6.2 years and DTSQs total treatment satisfaction score 21.7 ± 7.4. Percentages of responders (improvement in DTSQs total score ≥4 points) were 60.2% at month 3 and 75.2% at month 12. A decrease in perceived frequency of hyper/hypoglycemia and higher reduction in mean HbA1c and FPG from baseline to month 12 were also observed for responders vs. non-responders (Table).
Conclusion: The majority of insulin naïve PwT2D initiating Gla-300 reported meaningful increase in treatment satisfaction and decrease in perceived frequency of hyper/hypoglycemia, associated with decreased HbA1c.

Disclosure
S.B. Harris: Consultant; Abbott. Research Support; Boehringer-Ingelheim. Consultant; Dexcom, Inc. Advisory Panel; Eli Lilly and Company. Consultant; Eli Lilly and Company, Novo Nordisk, Sanofi. Research Support; Novartis AG. Consultant; Bayer Inc. N. Khan: None. A. Tirosh: Advisory Panel; Novo Nordisk. Speaker's Bureau; Eli Lilly and Company. Advisory Panel; Sanofi, GlaxoSmithKline plc, Medtronic. Consultant; Radella Pharmaceuticals. J. Kesavadev: None. H. Vargas-Uricoechea: Speaker's Bureau; Abbott, Sanofi-Aventis U.S. A. Roborel de Climens: Stock/Shareholder; Sanofi. H. Baghous: Other Relationship; Sanofi, Novo Nordisk. F.J. Snoek: Advisory Panel; Abbott, Sanofi, Eli Lilly and Company, Roche Diabetes Care. M.N. Mabunay: None. N. Grulovic: Employee; Sanofi. V. Corp dit Genti: Employee; Sanofi. J. Msihid: Employee; Sanofi. Stock/Shareholder; Sanofi. G.R. Galstyan: None.

Funding
Sanofi
210-OR: ADA Presidents' Select Abstract: Discovering Stimulatory State-Specific Type 2 Diabetes GWAS Mechanisms with Single-Cell Multiomics on iPSC-Derived Fibro-adipogenic Progenitor Cell Villages
Authors: VENTRESCA CHRISTA , VARSHNEY ARUSHI , ORCHARD PETER , TSAN YAO-CHANG , MONTEIRO DA ROCHA ANDRE , LAAKSO MARKKU , TUOMILEHTO JAAKKO , LAKKA TIMO A. , MOHLKE KAREN L. , BOEHNKE MICHAEL , SCOTT LAURA , KOISTINEN HEIKKI A. , COLLINS FRANCIS S. , HERRON TODD J. , BIELAS STEPHANIE , PARKER STEPHEN 
Published: June 2024

Variants at type 2 diabetes (T2D) and related trait genome wide association study (GWAS) signals may impact T2D risk via skeletal muscle, a primary insulin responsive tissue. We previously generated in vivo single nucleus (sn-)multi-omics (RNA+ATAC) profiling of 286 skeletal muscle biopsies from the FUSION Tissue Biopsy Study and found that a subset of GWAS signals colocalize with cell-type specific e/caQTL active in fibro-adipogenic progenitors (FAPs). In a subset of 50 of these individuals, we derived induced pluripotent stem cell (iPSC) lines from fibroblasts. We hypothesize that FAPs can be differentiated from iPSC lines to investigate the impact of FAP-specific molecular mechanisms on T2D risk and T2D-related traits. Here, we demonstrate that FAPs can be derived from iPSC lines for deep molecular phenotyping. These iPSC-derived FAPs display FAP morphology, expression of FAP marker genes, and loss of expression of pluripotency markers. Over the course of the differentiation, PODXL, a pluripotency marker, drops from 99.1% of cells expressing this down to 0.19%, based on flow cytometry analysis. Meanwhile, expression of NT5E, a FAP marker, increases from 0.095% of cells up to 98.9%. We performed a time course analysis with 10 independent iPSC lines, all multiplexed into a cell village, to explore the trajectory from iPSCs to FAPs. A logistic regression between cell type specific chromatin accessibility peaks of the iPSC-FAPs and skeletal muscle cell types reveals the chromatin accessibility profile of iPSC-FAPs to be very similar to in vivo FAPs, indicating that the differentiated FAPs can be used as a model system to characterize FAPs. We are currently upscaling the experiment to 40 iPSC-derived FAP samples to investigate stimulatory state specific genetic regulatory effects (e/caQTL) within T2D pathways. We will validate our results by performing CRISPRi knockdown and CRISPRa activation on a subset of nominated loci.

Disclosure
C. Ventresca: None. A. Varshney: None. P. Orchard: None. Y. Tsan: None. A. Monteiro da rocha: None. M. Laakso: None. J. Tuomilehto: Stock/Shareholder; Orion Pharma, Aktivolabs, Digostics. T.A. Lakka: None. K.L. Mohlke: None. M. Boehnke: None. L. Scott: None. H.A. Koistinen: Other Relationship; AstraZeneca, Novo Nordisk. F.S. Collins: None. T.J. Herron: None. S. Bielas: None. S. Parker: Research Support; Pfizer Inc.

Funding
National Institutes of Health (1F31DK135342-01A1); National Institutes of Health (R01DK117960); University of Michigan (UM1DK126185)
2101-LB: Restoring ß-Cell Mass and Identity in KATP-Induced Neonatal Diabetes—The Synergy of Autophagy and Intermittent Fasting
Authors: CASTELBLANCO ESMERALDA , REMEDI MARIA S. , RAMIREZ SOTERO IRVING 
Published: June 2024

Pancreatic β-cell dysfunction is central to diabetes pathogenesis. Autophagy, which degrades damaged organelles and proteins, has emerged as a key player in β-cell function, but its role in preserving β-cell mass and identity remains poorly understood. Here, we investigated the interplay between autophagy and β-cell identity in the tamoxifen-inducible KATP gain-of-function (GOF) mouse model of neonatal diabetes. As expected due to the expression of overactive channels, non-obese KATP-GOF mice developed severe diabetes due to a lack of insulin secretion. As diabetes progressed, they showed increased β-cell endoplasmic reticulum (ER) stress and a decline in β-cell mass and identity, accompanied by dysregulated autophagy with impaired autophagic flux and autophagosome accumulation. KATP-GOF mice showed altered liver architecture with hepatocytes containing lipid droplets, but no changes in liver autophagy markers were observed. Strikingly, KATP-GOF mice exposed to alternate-day intermittent fasting (IF) showed improved glycemic control and glucose tolerance, mitigation of β-cell ER stress and mitochondrial abnormalities, and restored β-cell mass and identity. IF KATP-GOF mice demonstrated normalization of β-cell autophagic flux and reduction of autophagosome accumulation, accompanied by restored liver architecture and enhanced expression of genes involved in gluconeogenesis and lipolysis. These studies demonstrate, for the first time, impaired autophagy in non-obese KATP-induced diabetes and suggest that IF preserved β-cell identity via improved autophagy in the absence of body-weight loss or insulin secretion, implying similar mechanisms occurring in different forms of diabetes. These findings not only increase our understanding of the mechanisms underlying β-cell demise in diabetes but also offer insights into therapeutic strategies aimed at preserving functional β-cell mass.

Disclosure
E. Castelblanco: None. M.S. Remedi: None. I. Ramirez Sotero: None.

Funding
National Institutes of Health (R01DK123163)
2102-LB: Dysregulation of Cholesterol Homeostasis Is an Early Signal of Beta-Cell Proteotoxicity Characteristic of Type 2 Diabetes
Authors: GURLO TATYANA , LIU RUOSHUI , YANG XIA , BLENCOWE MONTGOMERY , BUTLER PETER C. 
Published: June 2024

Type 2 diabetes (T2D) is a common metabolic disease due to insufficient insulin secretion by pancreatic beta cells in the context of insulin resistance. Islet molecular pathology reveals a role for protein misfolding in beta cell dysfunction and loss with islet amyloid derived from islet amyloid polypeptide (IAPP), a protein co-expressed and co-secreted with insulin. The most toxic form of misfolded IAPP is intracellular membrane disruptive toxic oligomers present in beta cells in T2D and in beta cells of mice transgenic for human IAPP (hIAPP). Prior work revealed a high degree of overlap of transcriptional changes in islets from T2D and pre-diabetic 9-10-week-old mice transgenic for hIAPP with most changes being pro-survival adaptations and therefore of limited therapeutic guidance. The aim of the study was to investigate islets from hIAPP transgenic mice at an earlier age (6 weeks) to screen for potential mediators of hIAPP toxicity that precede predominance of pro-survival signaling. Islets from 6-week-old hIAPP mice and wild-type mice were subjected to bulk RNA sequencing analysis followed by differential expression analysis, and then compared to prediabetic 9-10 weeks-old mouse islets. Analysis of intracellular lipid distribution in islets was performed in pancreas sections and isolated islets. hIAPP misfolding induced early suppression of cholesterol synthesis and trafficking along with aberrant intra-beta cell cholesterol and lipid deposits. The abnormality in lipid homeostasis mediated by hIAPP misfolding align with comparable lipid deposits present in beta cells in T2D and increased vulnerability to develop T2D in individuals taking medications that suppress cholesterol synthesis.

Disclosure
T. Gurlo: None. R. Liu: None. X. Yang: None. M. Blencowe: None. P.C. Butler: Consultant; Vertex Pharmaceuticals Incorporated.

Funding
Larry L. Hillblom Foundation (2014-D-001-NET, 2021-D-004-SUP)
223-OR: Characterization of Regenerative Adult Islet Progenitor Cells from Human Pancreatic Tissue
Authors: DARDEN CARLY , KUNCHA JAYACHANDRA , MATTKE JORDAN D. , LAWRENCE MICHAEL C. , NAZIRUDDIN BASHOO 
Published: June 2024

Introduction & Objective: To identify and characterize expandable islet progenitor cells in human pancreatic tissue capable of regenerating islet organoids in vitro and in vivo.
Methods: Single cell RNA-sequencing was performed on dissociated pancreatic tissue from normal human pancreas and pancreatitis to identify islet progenitor cell populations, gene signatures, and unique markers expressed in vitro and in vivo. Islet progenitors were validated by QPCR and flow cytometry. Islet progenitor cells were isolated by flow-sorting and expanded to produce functional islet-like organoids. Undifferentiated islet progenitor cells and differentiated islet-like organoids were transplanted into STZ-induced diabetic nude mice to assess islet function. Grafts and native pancreas were assessed for expression of islet organoids.
Results: A total of 9807 cells were analyzed and multiple cell types were identified and matched to pancreatic cell subtypes, including activated stellate cells (86%), ductal cell (9.2%), beta cell (3.8%), and alpha cell (0.5%). Human islet progenitors were identified expressing stem-cell marker Procr, immature beta-cell marker CD9, and islet endocrine progenitor marker RGS16. These Procr+, CD9+, RGS16+ cells also expressed ductal epithelial, stellate, and mesenchymal cell markers indicative of epithelial-mesenchymal transition. Undifferentiated islet progenitor cells could migrate to the damaged mouse pancreas to produce human islet organoids in vivo within 28 days post-transplantation. Islet progenitor cells could be differentiated to produce and release insulin and glucagon in vitro and restore islet cell function in vivo.
Conclusion: Procr+, CD9+, RGS16+ islet progenitor cells expanded from human pancreatic tissue retain regenerative properties to produce functional islet cell organoids in vitro and in vivo for potential use in islet cell replacement therapy or strategies to regenerate islet cells in the native pancreas.

Disclosure
C. Darden: None. J. Kuncha: None. J.D. Mattke: None. M.C. Lawrence: None. B. Naziruddin: None.
241-OR: Health-Related Quality of Life and Burden of Type 1 Diabetes and Severe Hypoglycemia in Adult Continuous Glucose Monitor Users
Authors: BOATENG-KUFFOUR ADRIANA , CALLAHAN PATRICK , CHANDARANA KEVAL , BARRY DIANNE , CHEN LIANG , KELLY CAITLIN S. , NGUYEN HUYEN , CHAPMAN KATHERINE S.M. , CORNELIUS EMILEE M. , WOLF WENDY , POLONSKY WILLIAM H. 
Published: June 2024

Introduction & Objective: Continuous glucose monitors (CGM) help manage glycemic control, but severe hypoglycemic events (SHE) remain challenging for adults with T1D. SHE may also reduce health-related quality of life (HRQoL) and increase humanistic and economic burden. Published literature on HRQoL in adult CGM users with T1D is limited.
Methods: Adult (≥18 years) CGM users from the T1D Exchange Registry completed an online self-report survey with SHE experiences, impaired awareness of hypoglycemia (IAH, Modified Gold Score), and patient-reported outcome measures (PROM). SHE cohorts were created based on SHE frequency in the past 12 months and presence/absence of IAH: problematic SHE, single SHE/no IAH, undetermined IAH, and no SHE (defined in the Table). Descriptive results (mean, SD) of PROM are reported overall and by SHE cohort.
Results: Overall, participants (N=1,847) were 45.9 years old, had T1D for 29 years (SD 15.1), and reported 1.8 SHE (SD 13.3) in the past year. Variability in patient-reported burden and HRQoL impairment was observed across cohorts. The problematic SHE cohort reported the highest mean burden and impairment (Table).
Conclusion: Despite advanced diabetes technology utilization, individuals with T1D and SHE experience substantial negative impacts of HRQoL, highlighting the significant unmet need in this patient population.

Disclosure
A. Boateng-Kuffour: Employee; Vertex Pharmaceuticals Incorporated. P. Callahan: Employee; Vertex Pharmaceuticals Incorporated. K. Chandarana: Employee; Vertex Pharmaceuticals Incorporated. Stock/Shareholder; Vertex Pharmaceuticals Incorporated, Novo Nordisk, Eli Lilly and Company. D. Barry: Employee; Vertex Pharmaceuticals Incorporated. L. Chen: None. C.S. Kelly: None. H. Nguyen: None. K.S.M. Chapman: None. E.M. Cornelius: None. W. Wolf: None. W.H. Polonsky: Research Support; Abbott Diagnostics, Dexcom, Inc. Consultant; Eli Lilly and Company, Sanofi. Advisory Panel; embecta. Consultant; Vertex Pharmaceuticals Incorporated. Advisory Panel; 9am Health.

Funding
Vertex Pharmaceuticals Incorporated
247-OR: Blood Monocyte Phenotype Predicts Cardiovascular Risk in Type 2 Diabetes
Authors: VENTECLEF NICOLAS , JULLA JEAN-BAPTISTE , GAUTIER JEAN-FRANÇOIS , POTIER LOUIS 
Published: June 2024

Background: Diabetes is a major risk factor for atherosclerotic cardiovascular diseases with a 2-fold higher risk of cardiovascular events in people with diabetes compared with those without. Circulating monocytes are inflammatory effector cells involved in both type 2 diabetes (T2D) and atherogenesis.
Methods: We investigated the relationship between circulating monocytes and cardiovascular risk progression in people with T2D, using phenotypic, transcriptomic, and metabolomic analyses. cardiovascular risk progression was estimated with coronary artery calcium score in a cohort of 672 people with T2D.
Results: Coronary artery calcium score was positively correlated with blood monocyte count and frequency of the classical monocyte subtype. Unsupervised k-means clustering based on monocyte subtype profiles revealed 3 main endotypes of people with T2D at varying risk of cardiovascular events. These observations were confirmed in a validation cohort of 279 T2D participants. The predictive association between monocyte count and major adverse cardiovascular events was validated through an independent prospective cohort of 757 patients with T2D. Integration of monocyte transcriptome analyses and plasma metabolomes showed a disruption of mitochondrial pathways (tricarboxylic acid cycle, oxidative phosphorylation pathway) that underlined a proatherogenic phenotype.
Conclusions: In this study, we provide evidence that frequency and monocyte phenotypic profile are closely linked to cardiovascular risk in patients with T2D. The assessment of monocyte frequency and count is a valuable predictive marker for risk of cardiovascular events in patients with T2D.

Disclosure
N. Venteclef: None. J. Julla: Speaker's Bureau; Lilly Diabetes, Novo Nordisk. Board Member; Sanofi. J. Gautier: Board Member; Sanofi. Other Relationship; Sanofi. Speaker's Bureau; AstraZeneca, Novo Nordisk. Research Support; Novo Nordisk. Advisory Panel; Pfizer Inc. L. Potier: Research Support; Abbott. Consultant; AstraZeneca, Bayer Inc., Eli Lilly and Company, Novo Nordisk, Sanofi, Boehringer-Ingelheim.