bluuebunny/crossref_metadata_2025_split huggingface dataset:
224-OR: Effect of PD-L1 Overexpression on Autoreactive T-Cell Responses toward Stem Cell–Derived Beta Cells
Authors: BARRA JESSIE , CASTRO-GUTIERREZ ROBERTO , SANTINI-GONZALEZ JORGE , MANSON CAMERON , WIDENER ADRIENNE , LANDRY LAURIE , NAKAYAMA MAKI , PHELPS EDWARD , RUSS HOLGER A. 
Published: June 2024

Introduction & Objective: The recent successful large-scale generation of human stem cell (hPSC)-derived β-like cells (sBC) provides an abundant cell source for replacement therapy efforts to treat patients suffering from autoimmune diabetes. However, even in the presence of systemic immune suppression, transplanted sBC, like primary human islets, are susceptible to allogeneic and recurrent autoimmune attack. Different strategies leveraging genome engineering technologies have been tested to achieve localized immune protection for β-cell grafts. Increased expression of the immune checkpoint inhibitor PD-L1 at the graft site has shown promise in reducing either xenogeneic or allogeneic immune responses in preclinical animal models. However, whether PD-L1 expression can protect sBC grafts from recurring autoimmunity is poorly understood, largely due to the absence of appropriate model systems.
Methods: Here we adoptively transferred diabetogenic mouse T cells from NOD mice expressing human HLA class I into immunodeficient NSG mice bearing HLA matched sBC grafts as a novel model of human autoimmunity in vivo.
Results: Using this novel autoimmune in vivo model, sBC grafts that were genetically modified to express high levels of PD-L1 exhibit greater survival compared to unmodified control grafts but were also lost within 3 weeks due to robust T cell infiltration. Interestingly, we also observed destruction of HLA class 1 knockout sBC grafts, suggesting a potential and important role for autoreactive CD4+ T cells. We are currently expanding our findings using HLA matched in vitro co-culture assays and performing additional transplantation studies with purified diabetogenic CD8+ T cells.
Conclusion: Collectively, our data suggest that PD-L1 expression on sBC alone might not provide sufficient protection from recurring autoimmunity, unlike allogenic responses, and highlights the need for better model systems to interrogate human autoimmunity in a matched manner.

Disclosure
J. Barra: None. R. Castro-Gutierrez: None. J. Santini-Gonzalez: None. C. Manson: None. A. Widener: None. L. Landry: None. M. Nakayama: None. E. Phelps: Research Support; Immunocore, Ltd, MESO SCALE DIAGNOSTICS, LLC. H.A. Russ: Advisory Panel; Sigilon Therapeutics, Inc. Consultant; Minutia. Advisory Panel; Prellis Biologics. Consultant; Eli Lilly and Company.

Funding
NIH/NIDDK (R01DK132387); JDRF (2-SRA-2023-1313-S-B); JDRF (2-SRA-2023-1367-S-B); Diabetes Research Connection (grant #49)
243-OR: Characterization of Behavioral Patterns Involving Hypoglycemic Episodes in Type 1 Diabetes—Results from the T1DEXI Study Cohort
Authors: CYPRIS LENA E. , GARCIA-TIRADO JOSE F. 
Published: June 2024

Hypoglycemia in T1D is considered the main factor limiting tight glycemic control, with well-documented consequences in morbidity, mortality, and economic cost. Hypoglycemia-related simultaneous glucose-insulin time series clustering has not been examined before. We analyzed data from 392 individuals (CSII and AID) from the T1DEXI dataset. A multivariate time series clustering algorithm on normalized CGM and IOB data was developed, using dynamic time warping and a threshold-based algorithm to discern postprandial (PP) glucose-insulin profiles related to hypoglycemic episodes. ANOVAs and independent t-tests were used to assess within- and between-profile differences of the AUC of IOB (total, basal rate, and bolus), respectively. 7500 (19.4%) out of 38744 3-hour PP periods had at least one hypoglycemic episode. Within this subset, we identified sixteen distinct glucose-insulin clusters, categorized into four behavioral profiles, P1-P4 (Fig. 1). Late PP hypoglycemia (P1), accounting for 34.8% of all episodes, is associated with a higher total basal rate and bolus than the desirable decision-making (P3) and late PP hyperglycemia (P4) profiles; all comparisons with p&lt;0.001.
In conclusion, new glucose-insulin behavioral profiles may provide actionable insight at the population and individual levels to minimize hypoglycemia risk.

Disclosure
L.E. Cypris: None. J.F. Garcia-Tirado: Research Support; Dexcom, Inc. Other Relationship; Dexcom, Inc.
248-OR: Pro-B-Type Natriuretic Peptide Strongly Predicts Major Cardiovascular Events and Cardiovascular Mortality in Cardiovascular Disease Patients with NAFLD as Well as in Those without NAFLD
Authors: PLATTNER THOMAS , LARCHER BARBARA , MADER ARTHUR , VONBANK ALEXANDER , LEIHERER ANDREAS , MUENDLEIN AXEL , DREXEL HEINZ , SAELY CHRISTOPH H. 
Published: June 2024

Elevated pro-B type natriuretic peptide (pro-BNP) has been found to be a marker of cardiovascular risk in several risk populations. However, its power to predict MACE and cardiovascular death in patients with the combination of established cardiovascular disease (CVD) and non-alcoholic fatty liver disease (NAFLD) is unclear. We therefore enrolled 639 patients with established CVD, 385 with angiographically proven stable CAD and 254 with sonographically proven peripheral artery disease. Prospectively, cardiovascular events were recorded over 9.6±4.3 years. At baseline, pro-BNP was not significantly different in patients with NAFLD (n=317) than in those who did not have NAFLD (827±2360 vs. 847±2754 pg/ml; p=0.236). During follow-up, 243 patients suffered MACE and 138 cardiovascular death; event rates of MACE and cardiovascular death were significantly higher in patients with NAFLD than in subjects without NAFLD (43.9 vs. 33.2%; p=0.006 and 26.3% vs. 17.6%; p=0.008, respectively). Pro-BNP predicted MACE and cardiovascular death in the total study population, with standardized adjusted hazard ratios (HR) of 1.55 [1.40-1.71]; p&lt;0.001 and 1.56 [1.41-1.74]; p&lt;0.001. Further, pro-BNP predicted MACE and cardiovascular death in patients with NAFLD (HRs 1.78 [1.54-2.07]; p&lt;0.001 and 1.80 [1.53-2.12]; p&lt;0.001, respectively) as well as in those who did not have NAFLD (HRs 1.47 [1.28-1.68]; p&lt;0.001 and 1.51 [1.30-1.75]; p&lt;0.001, respectively). Interaction terms pro-BNP x NAFLD were not significant for MACE and CD (p=0.151 and p=0.197, respectively), indicating that the power of pro-BNP to predict MACE and cardiovascular death did not differ significantly between CVD patients with NAFLD and those who did not have NAFLD. We conclude that pro-BNP strongly predicts major cardiovascular events and cardiovascular death in CVD patients with NAFLD as well as in those without NAFLD.

Disclosure
T. Plattner: None. B. Larcher: None. A. Mader: None. A. Vonbank: None. A. Leiherer: None. A. Muendlein: None. H. Drexel: None. C.H. Saely: None.
255-OR: The Relationship between Daytime Child Glycemic Levels and Parent Fear of Hypoglycemia
Authors: MONZON ALEXANDRA , MCDONOUGH RYAN , PATTON SUSANA R. 
Published: June 2024

Introduction: Hypoglycemia (i.e., &lt;70mg/dL) is a common and dangerous complication for youth with type 1 diabetes (T1D). Many parents report significant fear related to potential hypoglycemia and may engage in maladaptive avoidance behaviors to maintain higher glucose levels. Previous studies have mainly relied on HbA1C when examining the relationship between glucose patterns and parent fear of hypoglycemia (FoH). This study, however, examined if daytime child glucose levels, measured continuously, predicted parental FoH.
Methods: We recruited 165 caregivers of youth with T1D. We administered the Hypoglycemia Fear Survey for Parents and collected 14 days of glucose data from their child’s personal continuous glucose monitor. We conducted regression models with youth daytime glucose metrics as the independent variable and the parents’ FoH score as the dependent variable.
Results: Among caregivers, 89% self-identified as mothers with Mage= 41.76+6.52 years. For youth, 49% were female, Mage= 13.41+2.59 years, MT1D duration= 3.97+3.14 years, and MHbA1C= 7.81 + 1.43. Further, the average percent of glucose values below range= 2.5+3.2%, in range= 51.9+22.7%, and above range= 45.5+24.0%. Results revealed that youth’s percent of glucose values in the hypoglycemic range did not predict parents’ FoH score (p= 0.32). However, average daily glucose (β= 0.32, p&lt; 0.01) and glucose SD, or glycemic variability (β= 0.30, p&lt; 0.01), were significant predictors of parents’ FoH score.
Conclusions: Our results indicate that parents of youth with T1D may report higher FoH if they observe increased fluctuations in their child’s glucose levels, regardless of how often their child’s glucose levels are in the hypoglycemic range. The findings are important to clinicians who should consider screening for parental FoH to determine if glucose management should be a target for diabetes education or to provide strategies to help parents manage their specific worries related to hypoglycemia.

Disclosure
A. Monzon: None. R. McDonough: Speaker's Bureau; Provention Bio, Inc. S.R. Patton: None.
259-OR: Real-Time CGM Achieves Higher TIR among Pregnant Persons with GDM
Authors: VALENT AMY M. , HUERTAS-PAGAN CHRISTIAN , WARD LUCY , RICKERT MICHAELA C. , RINCON MONICA 
Published: June 2024

Objective: Determine if real-time CGM increases glucose TIR compared to current standard care of GDM using self-monitoring blood glucose (SMBG; ~4 times per day).
Methods: Randomized (2:1), open label trial comparing real-time CGM to SMBG alone (with blinded CGM) to manage persons w/ GDM. Subjects with &lt;7 days of CGM data were excluded. Primary outcome: glucose TIR (60-140mg/dL) from randomization to delivery admission, comparing the CGM to SMBG group using two-sample t-tests.
Results: After exclusions, CGM n=74 & SMBG n=37 were randomized. There were no differences in demographics or gestational age at enrollment (265/7±55/7 vs 282/7±45/7 weeks, P=0.154). Significantly more glucose TIR in CGM than the SMBG arm; driven by higher daytime TIR & lower time &gt;140 mg/dL.
Conclusion: Persons with GDM achieve higher TIR using real-time CGM compared to SMBG alone.

Disclosure
A.M. Valent: None. C. Huertas-Pagan: None. L. Ward: None. M.C. Rickert: None. M. Rincon: None.
26-OR: METTL3 Is Essential for Exercise-Induced Cardiac Benefits in Diabetic Cardiomyopathy
Authors: WANG CHUNYAN , KANG JIAYI , SUGAI-MUNSON AYA , LIU ZIPENG , YAO WEIFENG , LI HAOBO 
Published: June 2024

Introduction & Objective: Exercise is an effective strategy in improving functional outcome in patients with diabetic cardiomyopathy (DiaCM). Methyltransferase like 3 (METTL3) is a N⁶-Methyladenosine (m6A) methyltransferase that has been implicated in cardiac development and disease. However, the role of METTL3 in DiaCM and in cardiac benefits of exercise on DiaCM, are unclear.
Methods: METTL3 floxed mice were injected with AAV9 encoding cardiac troponin-T (cTnT)-driven Cre recombinase to knockout METTL3 in cardiomyocytes (CMs). CM-specific METTL3 knockout mice, METTL3 floxed mice, and their wild-type littermates were subjected to either chow diet or high-fat (HFD) diet feeding and injection of streptozotocin (STZ) to induce DiaCM, followed by an eight-week exercise training. Some of the HFD+STZ mice were injected with AAV9 encoding cTnT-driven METTL3 to overexpress METTL3 in CMs.
Results: Mice treated with HFD+STZ displayed increased E/e’ and reduced global longitudinal strain, fractional shortening (FS) and cardiac METTL3 protein expression (p&lt;0.05 for all). Exercise improved diastolic (E/e’: Sedentary: -34.69 ± 1.25 vs Exercise: -29.48 ± 1.92, p=0.04, n=10), systolic function (FS: Sedentary: 54.24% ± 2.94 vs Exercise: 62.33% ± 2.12, p=0.04, n=10) and cardiac METTL3 protein expression. These effects of exercise were canceled by CM-specific METTL3 knockout (FS: Control: 57.17% ± 1.85 vs Knockout: 28.49% ± 3.98, p&lt;0.001, n=8). In HFD+STZ mice, CM-specific METTL3 overexpression improves diastolic and systolic (FS: Control: 44.31% ± 2.2 vs Overexpression: 50.56% ± 1.64, p=0.03, n=8) functions. METTL3 functioned by increasing m6A deposition in YBX-1 and promoted its transcription. CM-specific YBX-1 overexpression improved diastolic and systolic functions as well as reduced cardiac oxidative stress.
Conclusion: METTL3 regulates DiaCM through YBX-1 and is essential for cardiac benefits of exercise in DiaCM. METTL3 is a promising therapeutic target for combating DiaCM.

Disclosure
C. Wang: None. J. Kang: None. A. Sugai-Munson: None. Z. Liu: None. W. Yao: None. H. Li: None.

Funding
National Institutes of Health (R21AG077040); American Heart Association (20CDA35310184)
260-OR: Differences in Adipocyte Expandability and Function between Healthy Pregnancies and Pregnancies Complicated by Gestational Diabetes Mellitus and Preeclampsia
Authors: LEES ZOE , SALT IAN , FREEMAN DILYS J. 
Published: June 2024

In obesity, failure of appropriate adipocyte expansion leads to ectopic fat deposition and systemic insulin resistance. Maternal obesity leads to higher incidence of gestational diabetes mellitus (GDM) and pre-eclampsia (PE), however adipocyte expansion is poorly characterised in pregnancy. This study aimed to compare adipocyte size, adipose tissue (AT) fibrosis, mRNA expression and insulin signaling in third trimester subcutaneous AT (SAT) and visceral AT (VAT) biopsies collected at Caesarean section from non-labouring GDM, PE and BMI- and age-matched healthy normotensive pregnant women (NT). SAT adipocyte diameter did not differ in complicated pregnancies compared to NT, however VAT diameter was higher in GDM (NT (n=30): 66.1±12.4 µm, GDM (n=30): 77.6±11.7 µm, p=0.001) but not PE (NT (n=7): 69.8±11.2 µm, PE (n=7) 72.4±8.9 µm, p=0.65). There was VAT hypertrophy in GDM (NT: 8.7±14.1% vs. GDM: 22.0±21.9% very large adipocytes, p=0.002) but not PE (NT: 13.7±21.3% vs. PE: 16.0±15.4%, p=0.67). Total fibrosis, assessed by picrosirius red staining of paraffin-embedded AT, was lower in GDM (NT (n=19): 45±13% area, GDM (n=12): 22±8%, PE (n=18): 45±12%, p&lt;0.001); whereas pericellular fibrosis was higher in GDM (NT (n=18): 16±7%, GDM (n=12): 23±7%, PE (n=16): 15±4%, p=0.02). Higher HIF1A mRNA expression relative to PPIA was observed in GDM VAT (NT (n=11): 18.1±3.9%, GDM (n=11): 46.4±29.8%, PE (n=10): 25.3±3.6%, p=0.03). Lower insulin-stimulated phosphorylation of Akt was observed in GDM and PE SAT compared to NT by immunoblotting, yet did not reach significance (fold change in phospho-Akt:Akt NT (n=9): 43.1±70.1, GDM (n=7): 10.5±35.4, PE (n=5): 5.8±26.9 (p=0.19). These data suggest AT dysfunction in GDM with a phenotype typical of T2DM, whereas AT dysfunction in PE requires further elucidation.

Disclosure
Z. Lees: None. I. Salt: None. D.J. Freeman: None.

Funding
The University of Glasgow MVLS Doctoral Training Programme.
276-OR: Common Variant Polygenic Risk Score Influences Penetrance of Rare Variants in Monogenic Diabetes Genes
Authors: HUERTA ALICIA , MANDLA RAVI , NAGY DORKA , SZCZERBINSKI LUKASZ , MADSEN JESPER G.S. , COLE JOANNE B. , PORNEALA BIANCA , WESTERMAN KENNETH E. , LI JOSEPHINE H. , POLLIN TONI I. , FLOREZ JOSE C. , GLOYN ANNA L. , CEBOLA INES , MANNING ALISA , LEONG AARON , UDLER MIRIAM , MERCADER JOSEP M. 
Published: June 2024

Monogenic diabetes (MD) accounts for 0.4% of diabetes cases and is a paradigm for precision medicine. However, a high fraction of variants in MD genes reported in ClinVar have uncertain significance (VUS) or conflicting interpretations of pathogenicity (CIP), making potential MD diagnosis a challenge.We hypothesized that using large-scale rare variant association data for type 2 diabetes (T2D) risk could inform variant pathogenicity in the MD genes. We tested the association with T2D of 1,157 VUS/CIP in 52 established MD genes, in the largest rare-variant T2D genome-wide association study (51,256 T2D, 370,487 controls), using rare variant imputation and whole-genome sequencing. We identified 9 variants with intermediate penetrance (VIP) (odds ratio [OR] &gt; 5) in HNF1A, HNF4A, GCK, KCNJ11, WFS1, and POLD1, including a missense variant in HNF4A (p.Arg114Trp) associated with an 8-fold increased risk of diabetes (OR=8.2, [4.6-14], p=1.1×10-13). The penetrance of this variant was influenced by a T2D polygenic risk score (PRS), with carriers in the highest tertile of the PRS showing a risk (OR=18.3 [7.2-46.9], p=1.2×10-9) comparable to the effect of confirmed pathogenic HNF4A MODY variants (OR=19.3 [6.1-60.6], p=4.1×10-7), and carriers within the lower tertile of the PRS showing a much smaller risk (OR=2.6 [0.9,7.1], p=0.06). We also provided support for 21% of rare variants in the MD genes as benign based on their sufficient statistical power but null association with T2D. We show that large-scale association data can inform variant pathogenicity and that rare variant penetrance is influenced by PRS. Our findings support the existence of a spectrum between common and monogenic forms of diabetes rather than a categorical distinction.

Disclosure
A. Huerta: None. R. Mandla: None. D. Nagy: None. L. Szczerbinski: None. J.G.S. Madsen: None. J.B. Cole: None. B. Porneala: None. K.E. Westerman: None. J.H. Li: None. T.I. Pollin: None. J.C. Florez: Research Support; Novo Nordisk. Other Relationship; Novo Nordisk, AstraZeneca. A.L. Gloyn: Other Relationship; Genentech, Inc., Roche Pharmaceuticals. I. Cebola: None. A. Manning: None. A. Leong: Other Relationship; Merck & Co., Inc. M. Udler: Other Relationship; Up-To-Date. J.M. Mercader: None.

Funding
A.H.C. is supported by American Diabetes Association 11-23-PDF-35 grant. J.M.M. is supported by American Diabetes Association Innovative and Clinical Translational Award 1-19-ICTS-068
278-OR: Single-Nucleus Multiomics Profiling across 49 Individuals of Hispanic Ancestry Reveals Cell-Specific Skeletal Muscle Insulin-Responsive Gene Regulatory Signatures
Authors: LI BENJAMIN , VARSHNEY ARUSHI , TOVAR ADELAIDE , MANICKAM NANDINI , ORCHARD PETER , DEFRONZO RALPH A. , NORTON LUKE , PARKER STEPHEN 
Published: June 2024

More than 1,000 genome-wide association study (GWAS) signals have been identified for type 2 diabetes (T2D) and related metabolic traits. 90% of these map to non-coding regions. This has obfuscated efforts to establish mechanisms, partly because some variants may impact disease risk in stimulatory rather than basal contexts. To identify these context-specific mechanisms, we performed single-nucleus RNA and ATAC (snRNA, snATAC) profiling on paired vastus lateralis skeletal muscle biopsies before and after four-hour euglycemic hyperinsulinemic clamp in 49 individuals of Hispanic ancestry.
After rigorous quality control, we jointly clustered 62,642 RNA and 91,734 ATAC nuclei (154,376 total) with LIGER from 47 individuals. We identified 15 cell types ranging from type 1 fibers (30.4% of nuclei) to T cells (0.4% of nuclei). We analyzed type 2a fibers (27.1% of nuclei)-a representative, intermediate muscle fiber type. Of 11,228 tested genes, 31.4% (n=3,526) were significantly differentially expressed (FDR &lt;5%), using DESeq2 and adjusting for batch and individual factors. This dramatic transcriptomic response was significantly enriched (FDR &lt;5%) for response to hormone stimulus. Further, of 619,445 tested ATAC peaks, 7.3% (n=45,474) were significantly differentially accessible (FDR &lt;5%). Curiously, these differentially accessible regions (DARs) were enriched for T2D-related GWAS signals such as Stumvoll insulin sensitivity index (ISI) and waist-to-hip ratio, adjusted for BMI-but only in type 2a, and not other muscle fiber types. As an example, the rs12303671-T-G ISI GWAS signal lies in a type 2a-specific DAR, suggesting cell-type-specific regulation under stimulatory contexts at a disease-relevant GWAS signal. We are currently performing expression and chromatin accessibility QTL mapping. These analyses may nominate testable cell- and context-specific mechanisms of T2D risk in skeletal muscle.

Disclosure
B. Li: None. A. Varshney: None. A. Tovar: None. N. Manickam: Employee; 10x Genomics. P. Orchard: None. R.A. DeFronzo: Advisory Panel; AstraZeneca, Novo Nordisk, Boehringer-Ingelheim, Intarcia Therapeutics, Inc., Aardvark, Renalytix, Corcept Therapeutics, Alnylam Pharmaceuticals, Inc. Research Support; Boehringer-Ingelheim, AstraZeneca, 89bio, Inc., Amgen Inc., Medality, Corcept Therapeutics. Speaker's Bureau; AstraZeneca, Corcept Therapeutics, Renalytix. L. Norton: None. S. Parker: Research Support; Pfizer Inc.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK117960); National Institute of Diabetes and Digestive and Kidney Diseases (UM1DK126185); National Institute of General Medical Sciences (T32GM007863); National Human Genome Research Institute (T32HG000040)
283-OR: Adipose Stromal Cells Increase Insulin Sensitivity and Decrease Liver Gluconeogenesis in a Type 1 DM Mouse Model
Authors: CHEN LEE-WEI 
Published: June 2024

Diabetic ketoacidosis (DKA) is a serious complication of hyperglycemic emergency due to insulin deficiency through accelerated liver gluconeogenesis and glycogenolysis. DKA is most common in T1D. Islet cells or parncreas transplantation are alternatives to insulin injection in treating T1D but only suitble for a small number of patients. This study aimed to explore the effects and mechanism of adipose stromal cells on liver gluconeogenesis and insulin sensitivity in an insulin-depended type 1 diabetic mouse model. We obtained stromal vascular fractions from wild-type inguinal adipose tissue and transplanted into the peritoneal cavity of type I diabetic Akita (Ins2Akita) mice. We found that injection of 5x106 SVF cells from wild type adipose tissue significantly decreased the pro-inflammatory genes of TNF-α, IL-1β, IL-33, iNOS and DPP4 expression in liver and increase anti-inflammatory factor of IL-10 and Foxp3 expression in blood serum and liver tissue seven days after injection. Moreover, mRNA expression of g6pc and pck1 was significantly decreased in liver of Akita mice. Furthermore, administration of 5x106 or 1x107 SVF cells from wild type adipose tissue significantly increased insulin sensitivity, reduced fasting blood glucose, and restored glucose-responsive c-peptide expression when compared to the untreated Akita group. Together, these findings suggest that adipose-derived SVF cells suppress liver inflammation, regulate liver gluconeogenesis, and improve insulin sensitivity in a type 1 diabetes mellitus animal model. Therefore, adipose SVF cells might be a novel cellular therapeutic strategy to maintain liver gluconeogenesis steady in T1D.

Disclosure
L. Chen: None.

Funding
National Science Council Taiwan MOST 110-2314-B-A49A-540-MY3