bluuebunny/crossref_metadata_2025_split huggingface dataset:
211-OR: Massively Parallel Characterization of Thousands of Type 2 Diabetes–Associated Variants in Multiple Cell Types Nominates Candidate Causal Regulatory Alleles and Mechanisms
Authors: KOTHARI MADHAV , BHUIYAN REDWAN M. , ULIRSCH JACOB C. , DEWEY HANNAH , NERURKAR NIKETA , KURSAWE ROMY , KALES SUSAN , BERENZY DANIEL , TEWHEY RYAN , STITZEL MICHAEL L. 
Published: June 2024

Genome-wide association studies have identified &gt;500 loci associated with type 2 diabetes (T2D). Most of these variants are in non-coding regions and may play regulatory roles, but they have not been comprehensively tested for this role. Here, we assessed transcriptional regulatory activity of 6,109 fine-mapped variants in 204 T2D-associated loci from DIAGRAM, UKBB, and BBJ cohorts using massively parallel reporter assays (MPRA) in HepG2 hepatocytes, SGBS (pre)adipocytes, and EndoC-βH3 beta-cells, three cell types implicated in T2D pathophysiology, as well as K562, SK-N-SH, and A549 cells.
MPRA identified 946 expression-modulating variants (emVars) in a T2D-relevant cell type. 277 T2D-associated emVars in 81 loci overlapped an open chromatin region in T2D-relevant cell types; precision recall analysis revealed precision of 0.83 and recall of 0.3 for identifying high posterior inclusion probability (PIP &gt; 0.9) variants using this integrated information. Importantly, MPRA nominated as high priority candidate causal variants a single functional variant at 45 loci, the majority of which were incompletely resolved using fine-mapping (PIP&gt;0.9 (n=6), PIP 0.5-0.9 (n=3) and PIP&lt; 0.5 (n=36)). These include well-characterized variants such as rs11257655 (CDC123/CAMK1D locus), whose risk allele increases MPRA activity via enhanced FOXA2 binding, and new variants like rs528350911 (PIP 0.99), whose T2D risk allele decreases MPRA activity through predicted disruption of a GATA binding site, and rs917195 (PIP 0.96), whose T2D risk allele in the CRHR2 locus is predicted to disrupt binding of RFXs. Together, this study provides new insights into the regulatory effects of non-coding T2D-associated variants across multiple cell types and nominates a set of high-confidence candidate causal variants for 39.7% of T2D-associated loci tested.

Disclosure
M. Kothari: None. R.M. Bhuiyan: None. J.C. Ulirsch: Employee; Illumina Inc. H. Dewey: Stock/Shareholder; Moderna, Inc. N. Nerurkar: None. R. Kursawe: None. S. Kales: None. D. Berenzy: None. R. Tewhey: None. M.L. Stitzel: None.

Funding
Opportunity Pool Funding (UM1DK105554); NIDDK (R01DK118011); The Jackson Laboratory Director's Innovation Fund NHGRI (UM1HG009435)
220-OR: Reduced Apolipoprotein C-I Truncation Is Associated with Worsening of Insulin Resistance and Risk of Diabetes in Persons with Impaired Glucose Tolerance
Authors: KOSKA JURAJ , NEDELKOV DOBRIN , BILLHEIMER DEAN , SCHWENKE DAWN , REAVEN PETER 
Published: June 2024

Apolipoprotein C-I (apoC-I) is key regulator of triglyceride and HDL metabolism. It appears as native and truncated posttranslational proteoforms. Higher truncated-to-native apoC-I proteoforms ratio (C-I’/C-I) is associated with favorable cardiometabolic risk profile. Whether apoC-I proteoform composition is associated with declines in insulin sensitivity and secretion, and incident diabetes is unknown. Plasma apoC-I proteoforms (by mass spectrometry immunoassay) were measured at baseline in 524 participants of the ACT NOW, a placebo-controlled study of pioglitazone in impaired glucose tolerance. Baseline and follow-up indices of insulin sensitivity (ISI, by Matsuda’s equation) and secretion (∆I/∆G calculated as AUCinsulin/AUCglucose) were derived from glucose and insulin concentrations during an OGTT. At baseline, C-I’/C-I correlated with both ISI and ∆I/∆G (r=0.26; r=-0.17, both p&lt;0.0001). Prospectively, higher baseline C-I’/C-I was associated with higher ISI (by 9.4% [95% CI: 4.1, 13.9] per 1 SD; follow-up adjusted for baseline, time of follow-up and treatment group), lower ∆I/∆G (-4.9% [-7.7, -1.0]) and reduced diabetes risk (n=59 events, HR 0.75 [95%CI: 0.57-0.98]). The C-I’/C-I associations with insulin sensitivity and diabetes were independent of diabetes risk factors (e.g., BMI) and plasma lipids (HR 0.72 [0.54-0.96]). The C-I’/C-I association with diabetes was attenuated after adjusting for change (follow-up adjusting for baseline) in ISI (HR 0.82 [0.61-1.11]) but not in ∆I/∆G (HR 0.67 [0.49-0.93]).
In conclusion, lower apoC-I truncation was associated, independently of typical diabetes risk factors, with decline in insulin sensitivity and increased risk of diabetes in individuals with impaired glucose tolerance. The association with incident diabetes appears to reflect favorable effects of apoC-I truncation on insulin sensitivity but not on insulin secretion.

Disclosure
J. Koska: None. D. Nedelkov: None. D. Billheimer: None. D. Schwenke: None. P. Reaven: Research Support; Dexcom, Inc.

Funding
National Institutes of Health (R24-DK090958); Takeda Pharmaceuticals
230-OR: Comparative Effectiveness of Semaglutide in Type 2 Diabetes—Year 2 Results of a Randomized Pragmatic Clinical Trial
Authors: BUSE JOHN B. , SOULE BENJAMIN P. , HARTY BRIAN J. , NORDAHL HELENE , CZIRAKY MARK J. , WILLEY VINCENT , SKIBSTED SIMON 
Published: June 2024

The objective of this study was to generate real-world evidence to support clinical decision-making by evaluating the long-term effects of once-weekly SC semaglutide (sema) in routine clinical practice. SEPRA was a 2-year open-label, randomized, pragmatic study (NCT03596450) comparing the effectiveness of sema vs physician’s choice standard of care (SOC; any antidiabetes drug excluding sema) when added to ≤2 OADs for treatment intensification in adults with T2D. Endpoints at 2 years included percentage of patients achieving HbA1c &lt;7%, %-point change in HbA1c from baseline, and change in body weight. Patients were randomized to receive sema (n=644) or SOC (n=634); 58% and 59% had recorded HbA1c levels at year 2, respectively. Baseline characteristics were comparable between groups (Table). Sema dosing at year 2 was 17.5% at 0.25 mg, 22.7% at 0.5 mg, and 27.5% at 1 mg (22.8% left the study, 8.9% were not on sema). Of patients receiving SOC at year 2, 37.4% were not receiving the treatment selected for intensification. At year 2, a significantly greater percentage of patients achieved HbA1c &lt;7% and numerically greater weight loss with sema vs SOC. A lower percentage of patients required further treatment intensification with sema vs SOC. No new safety concerns were identified. SEPRA demonstrated that sema is more effective and durable than SOC for T2D treatment intensification in routine clinical practice.

Disclosure
J.B. Buse: Other Relationship; Novo Nordisk. Consultant; Corcept Therapeutics. Research Support; Corcept Therapeutics, Dexcom, Inc., Insulet Corporation. Consultant; Alkahest, Anji Pharmaceuticals, Aqua Medical, Altimmune Inc., AstraZeneca, Boehringer-Ingelheim, CeQur, Eli Lilly and Company, embecta, GentiBio, Glyscend Inc., Mellitus Health, Metsera, Pendulum Therapeutics, Praetego, LLC, Stability Health, Terns Pharmaceuticals, Insulet Corporation, Vertex Pharmaceuticals Incorporated, vTv Therapeutics. Other Relationship; Medtronic. Stock/Shareholder; Glyscend Inc., Mellitus Health, Pendulum Therapeutics, Praetego, LLC, Stability Health. B.P. Soule: Employee; Novo Nordisk. B.J. Harty: Research Support; Novo Nordisk. H. Nordahl: Employee; Novo Nordisk. M.J. Cziraky: Other Relationship; Carelon Research. V. Willey: Other Relationship; Carelon Research. S. Skibsted: Employee; Novo Nordisk A/S.

Funding
Novo Nordisk Inc.
249-OR: Cystatin C Predicts Cardiovascular Events in Patients with Coronary Artery Disease Both among Patients with and among Those without Type 2 Diabetes
Authors: PLATTNER THOMAS , LARCHER BARBARA , MADER ARTHUR , VONBANK ALEXANDER , LEIHERER ANDREAS , MUENDLEIN AXEL , DREXEL HEINZ , SAELY CHRISTOPH H. 
Published: June 2024

Cystatin C is an established biomarker for renal function, and, given the close association of chronic kidney disease and cardiovascular disease might indicate new-onset or deteriorating cardiovascular disease. However, evidence for cystatin C as a predictor of cardiovascular events is limited and controversial. We therefore aimed at investigating the role of cystatin C as a predictor of future cardiovascular events in a high risk-cohort of patients with established coronary artery disease (CAD). We prospectively recorded cardiovascular events in 1053 patients with angiographically verified CAD over a mean follow-up of 9.5±5.3 years. At baseline, 352 patients had type 2 diabetes mellitus (T2DM) and 701 did not have T2DM. During follow-up, 560 (53.2%) of our patients suffered cardiovascular events. Cystatin C proved to be a strong and independent predictor for cardiovascular events in the total study cohort (standardized adjusted HR 1.22 [1.12-1.33], p&lt;0.001). When T2DM status was taken into account, cystatin C significantly predicted cardiovascular events both in patients with T2DM (HR 1.18 [1.04-1.35], p=0.011) and in nondiabetic patients (HR 1.23 [1.09-1.38], p&lt;0.001). We conclude that cystatin C predicts cardiovascular events in CAD patients both among those with and among those without T2DM.

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.
250-OR: Ceramides Surpass LDL-Cholesterol in Predicting Cardiovascular Mortality in Diabetic and Nondiabetic Patients
Authors: LEIHERER ANDREAS , MUENDLEIN AXEL , SAELY CHRISTOPH H. , PLATTNER THOMAS , LARCHER BARBARA , MADER ARTHUR , VONBANK ALEXANDER , LAAKSONEN REIJO , FRAUNBERGER PETER , DREXEL HEINZ 
Published: June 2024

The power of cardiovascular risk prediction is biased by comorbidities (including T2DM), medical treatment, and age, which impact cholesterol levels. Consequently, when applying LDL-C, the true cardiovascular risk of such confounded patients is often underestimated. However, it is not clear whether this constraint for risk prediction may likewise apply to other lipid risk markers in particular ceramides. In this observational cohort study, we recorded cardiovascular mortality for 16 years in 1195 patients. Their median age was 67 years, all of them had a high preexisting cardiovascular risk, 51% were taking statins, and 30% had T2DM. We found a U-shaped association between cardiovascular mortality and LDL-C. We thus stratified the population according to the vertex into high (≥150mg/dL) and lower LDL-C (&lt;150mg/dL) and also according to the T2DM status. A Cox regression analysis revealed that LDL-C failed to predict cardiovascular mortality in any subgroup. In contrast, no U-shaped but a linear association was found for ceramide-based markers. They were able to significantly predict cardiovascular mortality after stratification and even after multivariate adjustment. We thus propose that ceramide-based predictors rather than LDL-C should be applied for a more accurate cardiovascular risk prediction in diabetic and other high-risk patients.

Disclosure
A. Leiherer: None. A. Muendlein: None. C.H. Saely: None. T. Plattner: None. B. Larcher: None. A. Mader: None. A. Vonbank: None. R. Laaksonen: None. P. Fraunberger: None. H. Drexel: None.
257-OR: ADA Presidents' Select Abstract: The Effect of a Reduced-Energy Diet upon Maternal Weight and Pregnancy Outcomes in Women with Gestational Diabetes—The DiGest Trial
Authors: KUSINSKI LAURA C. , JONES DANIELLE , ATTA NOORIA , TURNER ELIZABETH , OUDE GRIEP LINDA M. , RENNIE KIRSTEN L. , DE LUCIA ROLFE EMANUELLA , SHARP STEPHEN , MURPHY HELEN R. , TAYLOR ROY , MEEK CLAIRE L. 
Published: June 2024

Background: Reduced-energy diets improve clinical outcomes in type 2 diabetes but are untested in gestational diabetes.
Methods: In a randomized, controlled, double-blind, whole-diet replacement trial (ISRCTN; 37866), women with gestational diabetes and BMI &lt;u&gt;&gt;&lt;/u&gt;25 kg/m2 were assigned (1:1) to receive a standard-energy (2000 kcal/day) (control) or a reduced-energy diet (1200 kcal/day) (intervention) until delivery, provided as weekly dietboxes containing 40% carbohydrate, 35% fat and 25% protein. Diagnosis used the National Institute of Health and Care Excellence (NICE) and interim UK Covid-19 criteria. Primary outcomes were maternal weight change (enrolment to 36wks) and offspring standardized birthweight. Secondary outcomes included maternal glycemia, treatment requirements and neonatal outcomes.
Results: 423 women were randomized at 29wks gestation; (n=211 control; n=212 intervention). Outcome data were available for 386 (91.3%) participants at 36wks and 358 (84.6%) at delivery. Intervention and control groups had similar maternal weight change to 36wks (mean 0.41(SD 4.25) vs 0.50(4.15) kg; baseline-adjusted difference intervention vs control -0.148 (95%CI -0.95 to 0.68); p=0.741). The intervention reduced requirements for short-acting (odds ratio 0.37 (0.14 to 0.97); p=0.044) and long-acting (0.38 (0.19 to 0.75); p=0.006) insulin therapy at 36wks. Offspring had similar standardized birthweight (0.44(1.03) vs 0.44(0.85) SDS; 0.004(-0.20 to 0.19) p=0.967) and comparable rates of large-for-gestational-age and neonatal intensive care admission. Serious adverse events and withdrawals were similar between groups. Weight loss in late pregnancy appeared safe.
Conclusions: The protocol did not result in differences in weight gain or primary outcomes between groups but the provision of a reduced energy diet was associated with a reduced requirement for insulin therapy.

Disclosure
L.C. Kusinski: None. D. Jones: None. N. Atta: None. E. Turner: None. L.M. Oude Griep: None. K.L. Rennie: None. E. De Lucia Rolfe: None. S. Sharp: Consultant; Shionogi & Co., Ltd. H.R. Murphy: Advisory Panel; Medtronic. Research Support; Abbott, Dexcom, Inc. Speaker's Bureau; Ypsomed AG, Eli Lilly and Company, Dexcom, Inc. R. Taylor: Advisory Panel; Fast800. C.L. Meek: Research Support; Dexcom, Inc.

Funding
Diabetes UK 17/0005712EFSD - NNF NNF19SA058974
282-OR: Nod1 Plays a Causal Role in Western-Diet–Induced Insulin Resistance and Glucose Intolerance in Microbiota-Deficient Conditions
Authors: M NIAZUR RAHMAN S , TSAKIRIDIS NICOLE , GARCIA MIGUEL A. , PARK JUNYOUNG , TAN YAO , GIACCA ADRIA 
Published: June 2024

Introduction & Objective: Recent studies revealed that nucleotide-binding oligomerization domain 1 (Nod1), an innate immunity receptor, mediates high fat diet (HFD) effects to induce insulin resistance and glucose intolerance. Circulating free fatty acids, acting as damage-associated molecular patterns (DAMPs), activate Nod1, which also senses gut microbiota-derived pathogen-associated molecular patterns (PAMPs), which are thought to be absorbed into the circulation as HFD changes gut permeability. While Nod1-null mice showed protection from HFD-induced insulin resistance, it remains unclear if this is driven by PAMPs or DAMPs. To investigate whether the protective effect of Nod1 deficiency is abolished without PAMPs, we studied Nod1 knockout (KO) in germ-free (GF) conditions, minimizing PAMP influence to reveal DAMP-induced Nod1 activation's role in insulin resistance and T2D.
Methods: We subjected GF or conventionally maintained C57BL/6 Nod1 KO and wild type (WT) mice to an 8-week high-fat, high-sucrose diet (HFSD) starting at 5 weeks of age to mimic Western diet-induced glucose intolerance and early diabetes. At 13 weeks, mice underwent intraperitoneal insulin tolerance tests (IPITT) and glucose tolerance tests (IPGTT) three days apart to assess insulin sensitivity and glucose tolerance.
Results: HFSD-induced insulin resistance in WT mice was alleviated by Nod1 deficiency in both non-GF and GF KO groups, as demonstrated by IPITT. In IPGTT, calculated insulin sensitivity and disposition index revealed HFSD-induced insulin resistance and attenuated β-cell function in WT mice, which was improved in Nod1KO mice, regardless of GF conditions.
Conclusion: Our data suggest DAMP mediated Nod1 activation as displayed by protection from HFSD induced insulin resistance and glucose intolerance in the absence of microbial PAMPs. These findings indicate that Western diet exerts its diabetogenic effects via Nod1 independent of gut microbiota.

Disclosure
S. Rahman: None. N. Tsakiridis: None. M.A. Garcia: None. J. Park: None. Y. Tan: None. A. Giacca: None.

Funding
Canadian Institutes of Health Research (CIHR 507485)
284-OR: Androgen-Dependent Tissue Factor Pathway Inhibitor-Regulating Protein (ADTRP) Regulates Macrophage Polarization and Migration to Adipocytes
Authors: MIN ONG SZE , HENG CHEW-KIAT , SENG LEE YUNG , SHU QIN OOI DELICIA 
Published: June 2024

Introduction: Macrophage infiltration into adipose tissue and their polarization from an anti-inflammatory M2 to pro-inflammatory M1 phenotype contributes to adipose tissue inflammation and the development of insulin resistance. Androgen-dependent tissue factor pathway inhibitor-regulating protein (ADTRP) is a recently discovered enzyme that breaks down a class of anti-inflammatory and anti-diabetic lipids in adipose tissues. Our study aims to investigate the effect of ADTRP on macrophage polarization and migration to adipocytes.
Methods: Mouse 3T3-L1 mature adipocytes were either transduced with AAV-ADTRP viral vectors or transfected with ADTRP siRNA to overexpress or knockdown ADTRP respectively. Conditioned media from the ADTRP-overexpressed and ADTRP-knockdown adipocytes were collected and treated to mouse RAW 264.7 macrophages. Lipopolysaccharide (LPS) was used to induce a pro-inflammatory state in the macrophages. Gene expression of macrophage M1 and M2 markers, including IL-1β, IL-6, TNF-α, iNOS, CD206 and IL-10 were measured using RT-qPCR. Macrophage migration assay was performed by co-culturing RAW 264.7 macrophages and 3T3-L1 adipocytes in a transwell system.
Results: Conditioned media from ADTRP-overexpressed adipocytes reduced the expression of anti-inflammatory M2 macrophage markers, CD206 and IL-10. On the contrary, conditioned media from ADTRP-knockdown adipocytes increased M2 markers in basal-state macrophages and decreased pro-inflammatory M1 markers, including IL-1β, TNF-α and iNOS, in LPS-stimulated macrophages. We also observed reduced macrophage migration towards ADTRP-knockdown adipocytes.
Conclusion: ADTRP regulates macrophage polarization and migration to adipocytes. Inhibition of ADTRP can be further studied as a possible strategy to reduce adipose tissue inflammation.

Disclosure
S. Ong: None. C. Heng: None. Y. Lee: None. D. Ooi: None.

Funding
Khoo Teck Puat - National University Children's Medical Institute research grant
288-OR: Inflammation-Induced Repression of Insulin Production by Targeting Its mRNA to Stress Granules via ADP-Ribosylation Machinery
Authors: SARKAR SOUMYADEEP , LI FANGJIA , ELLIOTT EMILY , RAMOS-RODRÍGUEZ MIREIA , YOU YOUNGKI , PASQUALI LORENZO , SUSSEL LORI , METZ THOMAS , MIRMIRA RAGHAVENDRA G. , NAKAYASU ERNESTO 
Published: June 2024

ADP-ribosylation has been shown to contribute to the development of Type 1 diabetes (T1D); however, the mechanism through which each component of the ADP ribosylation machinery contributes to the disease is poorly understood. Here, we aimed to investigate the contribution of specific ADP-ribosyltransferases in the development of T1D. We conducted comprehensive bioinformatics analyses on our previously published RNA-Seq, ATAC-Seq, H3K27ac-Seq, and proteomic data, followed by qPCR to investigate the regulation of ADP-ribosyltransferases in islets and β cells. We found that PARP12 expression was induced by both cytokines and omega-3 fatty acids, which are inducers and reducers of insulitis, respectively. RNAi depletion of PARP12, followed by proteomics analysis, revealed that a variety of stress granule proteins are regulated by PARP12. We performed fluorescence in situ hybridization, ADP-targeted RNA pulldown and pulse-chase proteomics to demonstrate that PARP12 ADP-ribosylates Insulin2 mRNA, targeting it to stress granules and blocking its translation. Stress granules are a characteristic feature of the integrative stress response and unfolded protein response in T1D. Here, we showed that PARP12 directs insulin mRNA to stress granules, impeding translation in response to cytokines. This may have implications for insulitis and T1D, such that an abrupt inhibition of insulin translation may amplify the stress and immune responses. These insights also provide avenues for therapeutic strategies aiming to improve insulin production.

Disclosure
S. Sarkar: None. F. Li: None. E. Elliott: None. M. Ramos-Rodríguez: None. Y. You: None. L. Pasquali: None. L. Sussel: None. T. Metz: None. R.G. Mirmira: None. E. Nakayasu: None.

Funding
National Institutes of Diabetes and Digestion and Kidney Disease (U01 DK127505, U01 DK127786 and R01 DK105588); HIRN Catalyst Award.
303-OR: Notch Signaling Inhibitor ADPO-002 Promotes Browning in Human Adipose Tissue Explants
Authors: RAISUL ABEDIN MUHAMMAD , FREIJE WILSON , OSWALT MALLORY A. , ACTON ANTHONY J. , CRAWFORD CHRISTOPHER B. , INMAN MARGARET M. , CONSIDINE ROBERT V. , DENG MENG 
Published: June 2024

Introduction: Transformation of white adipocytes into beige adipocytes, known as adipocyte browning, holds great promise for development of new therapeutics. Our previous studies have demonstrated nanoparticle-based delivery of dibenzazepine (ADPO-002), a γ-secretase inhibitor, induces browning of white adipose tissue in obese mice and in pigs. The current study aims to investigate the browning efficacy of ADPO-002 in human adipose tissue organ culture.
Methods: Abdominal subcutaneous and omental adipose tissue samples were collected from 24 subjects undergoing bariatric surgery (BMI 47.5±3.2 kg/m2, age 44.8±3.0 yr). The study was approved by the Ascension St. Vincent hospital IRB and all subjects provided informed consent. Tissues were cut into small pieces (~5-10 mg) using surgical scissors and cultured in medium M199 supplemented with 0.7 nM insulin and 10 nM dexamethasone. The treatment groups were DMSO (negative control), 50 µM forskolin (positive control), and 3, 10 and 30 µM ADPO-002. After 7-days treatment, total RNA was isolated, and RT-qPCR performed for the browning marker genes PRDM16 and PGC1A. Statistical significance was determined using Wilcoxon non-parametric paired t-test.
Results: In omental adipose tissue, 30 µM ADPO-002 significantly increased PRDM16 (429.1±149.3 vs 10.6±4.9 relative units, p=0.0020) and PGC1A (117.4±58.7 vs 58.5±30.2 relative units, p=0.0273) expression compared to the control group. In subcutaneous adipose tissue, 30 µM ADPO-002 significantly upregulated PRDM16 expression compared to control group (398.9±198.6 vs 22.06±6.28 relative units, p=0.0039).
Conclusion: These findings suggest that ADPO-002 treatment increases the expression of genes associated with increased mitochondrial expression. This finding supports the hypothesis that inhibition of γ-secretase can increase browning in human adipose tissue.

Disclosure
M. Abedin: Employee; Adipo Therapeutics LLC. W. Freije: None. M.A. Oswalt: None. A.J. Acton: None. C.B. Crawford: None. M.M. Inman: None. R.V. Considine: Research Support; Eli Lilly and Company, Adipo Therapeutics. M. Deng: Other Relationship; Adipo Therapeutics.

Funding
Indiana Innovation Voucher