bluuebunny/crossref_metadata_2025_split huggingface dataset:
2057-LB: Amino Acids Plasma Levels Do Not Correlate with Lean Mass Loss in Weight-Matched DIO Mice Treated with Survodutide or Semaglutide
Authors: KLEPAC-BARTL KATARINA , PEREIRA MAFALDA , ZIMMERMANN TINA , RIST WOLFGANG , BAJRAMI BESNIK , KLOECKENER TIM , KLEIN THOMAS 
Published: June 2024

Concomitant with beneficial fat mass loss, anti-obesity therapies lead to detrimental lean mass loss. It is known that amino acids are essential for muscle mass maintenance, growth, and repair. Moreover, it is recognized that glucagon promotes skeletal muscle wasting to supply amino acids for gluconeogenesis. Given that survodutide is a dual glucagon/glucagon like peptide 1 (GLP1) receptor agonist, and semaglutide acts solely as GLP1 receptor agonist, it could be hypothesized that survodutide leads to greater lean mass loss. Hence, we investigated plasma amino acids levels, body composition and energy expenditure (EE) of DIO (diet induced obesity) mice (8 mice/group) treated with either survodutide or semaglutide (both at 20 nmol/kg s.c. q.d. 3 weeks). Similar weight loss was achieved with survodutide (17.7±3.3%) and semaglutide (15.6±3.7%) vs. baseline, mean±SD, p&lt;0.0001. While the weight loss for semaglutide was due to its anorexigenic effect, survodutide additionally increased EE (p=0.0081, d21) due to its glucagon component. Both compounds showed ~40% decrease in fat mass (40.4±1.7% and 40.8±3.7%, mean±SD, survodutide and semaglutide respectively, p=0.0006 vs. vehicle). Importantly, lean mass loss was not significantly different when comparing survodutide (8.3±0.8%) and semaglutide (5.3±0.9%), mean±SD, p=0.9071. Interestingly, both essential and non-essential plasma amino acids were decreased when comparing semaglutide with survodutide: L-lysine decreased from 180.5±32.7 µmol/L to 90.3±16.6 µmol/L and L-alanine from 336.4±121.4 µmol/L to 117.8±32.2 µmol/L when comparing semaglutide and survodutide respectively, mean±SD. In summary, we showed that the glucagon component of survodutide increased EE and reduced plasma amino acids levels when compared to semaglutide. However, these results did not translate into a different lean mass loss, indicating that the reduction in plasma amino acids did not correlate with lean mass loss.

Disclosure
K. Klepac-Bartl: Employee; Boehringer-Ingelheim. M. Pereira: Employee; Boehringer-Ingelheim. T. Zimmermann: Employee; Boehringer-Ingelheim. W. Rist: Employee; Boehringer-Ingelheim. B. Bajrami: Employee; Boehringer-Ingelheim. T. Kloeckener: Employee; Boehringer-Ingelheim. T. Klein: Employee; Boehringer-Ingelheim.

Funding
Boehringer Ingelheim
2058-LB: DA-1726, a GLP1R/GCGR Dual Agonist, a Promising Approach in Obesity Treatment and Lipid Management
Authors: CHAE YUNA , JUNG IL-HUN , KIM TAE-HYOUNG , KIM MI-KYUNG , JIN LEE SU , HEON KIM HYUNG 
Published: June 2024

DA-1726 is being prepared for obesity Phase 1 clinical trials following IND approval. Our study focuses on the pharmacological effects of this novel oxyntomodulin analogue, which has been effective at improving lipid profiles and reducing weight in rodent models. In a prior study, DA-1726 showed a difference in improving lipid levels, despite similar weight loss to tirzepatide. Therefore, the direct lipid-regulating effect of DA-1726 was assessed in a hypercholesterolemia rat model compared with tirzepatide. As a result, DA-1726 was more effective than tirzepatide in suppressing the elevation of T-CHO ( -33.5%, -25.5% vs. control; P&lt;0.05) and LDL-C (-53.2%, -41.5% vs. control; P&lt;0.05). This differentiated impact is thought to arise from DA-1726's glucagon action, alongside its GLP-1 effect. We evaluated whether these differential effects could also be distinguished from drugs of the same class. In an obese mouse model, DA-1726 significantly lowered T-CHO (-67.7%, -49.6%; P&lt;0.05 vs. control) and TG (-49.5%, -41.2%; P&lt;0.05 vs. control) levels under conditions that resulted in similar weight loss effects (-31.9%, -25.4%; P&lt;0.05 vs. control) as survodutide. Besides, DA-1726 exhibited superior glucose lowering compared to survodutide (-54.7%, -30.4% vs. control; P&lt;0.05). Interestingly, despite the same mechanism of action, DA-1726-treated mice maintained similar levels of weight loss while consuming more food due to less dietary restriction in terms of appetite suppression. This effect might stem from DA-1726's GLP-1 and glucagon receptor activity ratio. DA-1726's glucagon action could further enhance energy expenditure (EE), and it is believed to have significantly increased the expression of EE-related genes in brown adipose tissue. In summary, it was confirmed that DA-1726 has differentiating characteristics in the competition of obesity treatment drugs with similar or identical mechanisms in its effect on improving cholesterol metabolism through glucagon action.

Disclosure
Y. Chae: None. I. Jung: None. T. Kim: None. M. Kim: None. S. Lee: None. H. Kim: Employee; NeuroBo Pharamceuticals Inc., Dong-A ST Co., Ltd.
2059-LB: NA-931, a Novel Triple IGF-1/GLP-1/GIP Incretin Receptor Agonist Reduces Body Weight and Improves Metabolic Profile in DIO Mice
Authors: TRAN LLOYD L. 
Published: June 2024

Background: Insulin-like Growth Factor 1 (IGF-1) plays a major part in fuel metabolism and regulation of body composition. GIP and GLP-1 have been shown to be effective for weight loss in non-diabetic patients with obesity when given as adjunctive therapy to diet and exercise. IGF-1 contributes to modulate glucagon secretion in which IGF-1 inhibits the ability of low glucose concentration to stimulate glucagon expression and secretion. NA-931 and its analogs, NA-932 and NA-933 (“NA-931 Compounds”) are metabolites of IGF-1. Acting as a receptor agonists targeting IGF-1, GLP-1 and GIP, NA-931 Compounds may provide enhanced therapeutic benefits.
Methods: Male diet-induced obese (DIO) mice were treated with daily subcutaneous injections of vehicle or one of novel triple IGF-1/GLP-1/GIP receptor agonists, NA-931, NA-932 and NA-933 (10 nmol/kg), for 14 days. Tirzepatide (10 nmol/ kg) as used as positive controls. Cohorts were then assessed for changes in BW, glucose, and lipids.
Results: Treatment with NA-931 Compounds resulted in reductions to BW (up to 26%, p&lt;0.0001), plasma glucose, plasma triglycerides, (up to 23% and 34%, respectively, p&lt;0.003 for each), and liver triglycerides (up to 46%, p&lt;0.05) compared to vehicle treatment. Weight loss effects in cohorts treated with NA-931 Compounds were comparable to those observed in tirzepatide-treated animals. In addition, liver lipid reductions were numerically greater among animals treated with the NA-931Compounds.
Conclusions: NA-931 and its analogs produced significant reductions in BW in DIO mice. Effect sizes were comparable to those observed in the tirzepatide control group. The NA-931 Compounds have been shown to produce desirable changes to lipid profile, suggesting global cardiometabolic benefit, represent a promising therapeutic approach to metabolic disorders such as obesity, type 2 diabetes, and non-alcoholic steatohepatitis. Additional research on these drug candidates is ongoing.
2081-LB: Efficacy and Safety of Semaglutide and Tirzepatide—A Comparative Analysis of Clinical Trials
Authors: TRAN LLOYD L. , TRAN ZUNG V. 
Published: June 2024

Introduction & Objective: In the past years, many obesity drugs were approved by the FDA and withdrawn. Semaglutide and tirzepatide are dominating the obesity market. We analyzed published phase 2/3 clinical trials to evaluate their efficacy and safety.
Methods: We selected 17 randomized phase 2/3 trials. Participants had obesity, diabetes, or both. Outcomes were baseline - post treatment differences: efficacy [body wt (kg/%), fasting glucose (mg/dL), HbA1c (%)]; safety [nausea, diarrhea, vomiting, constipation]; also age, %women, %white, sample size, and duration, To estimate pooled differences, we used bootstrapping, a machine learning/AI methodology to resample a dataset, generating a sampling distribution for statistical calculations. We estimated means, 95%CI of the differences by performing 10,000 resamples.
Results: Table 1: Across the 17 trials, semaglutide and tirzepatide lowered body by 8.5 and 10.7 %; fasting glucose by 29.8 and 51.8 mg/dL; HbA1c by 1.1 and 1.8%, respectively.
Conclusion: Semaglutide and tirzepatide are comparable in their efficacy and safety profile. Tirzepatide is better at reducing fasting glucose. Dual agonism at GIP and GLP-1R may contribute to the weight control effects of tirzepatide. At the ADA conference, Biomed introduces NA-931, a new oral drug with triple agonism at GIP/ GLP-1 and IGF-1 as an alternative treatment with less AEs for those with obesity and diabetes.

Disclosure
Z.V. Tran: Employee; Biomed Industries, Inc.
2086-LB: Islet Surface Modification with Bissulfosuccinimidyl Suberate Prevents the Instant Blood-Mediated Inflammatory Reaction—A Novel Approach
Authors: KALIVARATHAN JAGAN , BALAJI SARAVANAN PRATHAB , FUGLESTAD BRIAN A. , DHAKAL SOMA , MOHAMMED SHUJAUDDIN , LEVY MARLON F. , KANAK MAZHAR A. 
Published: June 2024

Introduction and Objectives: In pancreatic islet cell transplantation, the early graft loss is due to the instant blood-mediated reaction (IBMIR). Control of early inflammatory events could improve the efficacy of islet cell transplantation and may enhance the long-term function of transplanted islets. Nanoencapsulation of islets is an attractive method for masking cell features that can trigger inflammatory cascades. Bissulfosuccinimidyl suberate (BS3) is a homobifunctional crosslinker with NHS ester on each side to bind to amine groups. Herein, we investigated the effects of BS3-modified human islets in terms of viability, function, longevity of modification, and inflammatory properties.
Methods: The surface of the human islets was modified with BS3 in different concentrations (0.5, 1, and 5 mM). Visual coverage and efficiency of surface conjugation were determined by incorporating an Amine-PEG-Biotin conjugate. Surface-conjugated islets were then cultured for 7 days. Visualization of conjugation and viability determination was performed days 0, 1, 4, and 7 of culture. The impact of BS3 conjugation on islet function was determined by measuring the oxygen consumption rates and glucose-stimulated insulin secretion.
Results: BS3 modification was found to be exceptionally biocompatible. The viability and functional integrity of islets were not altered at 0.5 mM and 1 mM BS3, but viability showed a mild decline at 5 mM BS3. However, improved conjugation of the islet surface and good coverage were noted in the 5 mM BS3 group. The surface modification was maintained for all 7 days tested with minimal loss of coverage. The markers of inflammation were examined using qPCR analysis. BS3 treated group showed higher inhibition of all pro-inflammatory genes under strong cytokine cocktail exposure.
Conclusion: Surface modification of islets with BS3 is a safe and tolerable approach that may be used for localized drug delivery strategies.

Disclosure
J. Kalivarathan: None. P. Saravanan: None. B.A. Fuglestad: None. S. Dhakal: None. S. Mohammed: None. M.F. Levy: None. M.A. Kanak: None.

Funding
Department of Defense (W81XWH2210102)
2088-LB: The Downregulation of SCGN Induced by Lipotoxicity Promotes NLRP3-Mediated ß-Cell Pyroptosis
Authors: YANG JING , OUYANG SHUHUI , XIANG SUNMIN 
Published: June 2024

Background & Objective:Lipotoxicity exacerbates β-cell injury to contribute type 2 diabetes development. In lipotoxic conditions, secretagogin (SCGN), a protein highly expressed in islets, is found to undergo downregulation. Thus, we aim to explore the role of SCGN in lipotoxicity-induced β-cell injury.
Methods: MIN6 cells were exposed to ox-LDL to simulate lipotoxic damage. To modulate SCGN expression, the pcDNA3.1-Scgn vector or SCGN-specific siRNAs were utilized in MIN6 cells. To reinstate SCGN expression, we administered AAV-9 carrying SCGN in high-fat diets (HFD) mice. Cell viability was evaluated through MTT and PI assays, pyroptotic pathway was assessed using PCR, western blotting, and immunofluorescence techniques.
Results: Exposure to ox-LDL in vitro or an HFD in vivo decreases SCGN expression and actives pyroptosis. Restoring SCGN expression partially reverses this effect. The mechanism behind this involves the downregulation of SCGN facilitates ChREBP translocation from cytosol to the nucleus. This, ultimately, promotes TXNIP transcription, activating the NLRP3/Caspase-1 pathway.
Conclusion: Lipotoxicity-induced SCGN downregulation facilitates ChREBP translocation from the cytosol to nucleus. This, subsequently, triggers
TXNIP transcription, leading to the activation of NLRP3/caspase-1 pyroptotic pathway. Ultimately, resulting in pyroptotic pancreatic β-cells death.

Disclosure
J. Yang: None. S. Ouyang: None. S. xiang: None.

Funding
National Natural Science Foundation of China (82170852)
2097-LB: HNF4A and HNF1A Exhibit Tissue-Specific Target Gene Regulation in Pancreatic Beta Cells and Hepatocytes
Authors: TEO ADRIAN 
Published: June 2024

HNF4A and HNF1A encode transcription factors that are important for the development and function of the pancreas and liver. Mutations in both genes have been directly linked to Maturity Onset Diabetes of the Young (MODY) and type 2 diabetes (T2D) risk. To better define the pleiotropic gene regulatory roles of HNF4A and HNF1A, we generated a comprehensive genome-wide map of their binding targets in pancreatic and hepatic cells using ChIP-Seq. HNF4A was found to bind and regulate known (ACY3, HAAO, HNF1A, MAP3K11) and previously unidentified (ABCD3, CDKN2AIP, USH1C, VIL1) loci in a tissue-dependent manner. Functional follow-up highlighted a potential role for HAAO and USH1C as regulators of beta cell function. Unlike the loss-of-function HNF4A/MODY1 variant I271fs, the T2D-associated HNF4A variant (rs1800961) was found to activate AKAP1, GAD2 and HOPX gene expression, potentially due to changes in DNA-binding affinity. We also found HNF1A to bind to and regulate GPR39 expression in beta cells. Overall, our studies provide a rich resource for uncovering downstream molecular targets of HNF4A and HNF1A that may contribute to beta cell or hepatic cell (dys)function, and set up a framework for gene discovery and functional validation.

Disclosure
A. Teo: None.

Funding
NMRC OFYIRG18may040A*STAR Career Development Fund 2019 202D800020IMCB, A*STARFY2019 SingHealth Duke-NUS Surgery Academic Clinical Programme Research Support Programme GrantPrecision Medicine and Personalised Therapeutics Joint Research Grant 20192nd A*STAR-AMED Joint Grant Call 192B9002HLTRP/2022/NUS-IMCB-02Paris-NUS 2021-06-R/UP-NUS (ANR-18-IDEX-0001)OFIRG21jun-0097CSASI21jun-0006MTCIRG21-0071SDDC/FY2021/EX/93-A147FY 2022 Interstellar Initiative Beyond grantH22G0a0005HLCA23Feb-0031
2100-LB: GLP-1/GIP Receptor Agonist Evaluation Using cAMP Measurement in EndoC-ßH5 Human Beta Cells
Authors: TAURAND MARION , OLLEIK HAMZA , BLANCHI BRUNO 
Published: June 2024

EndoC-βH5 human beta cells are ready-to-use functional human pancreatic beta cells. They robustly and reproducibly secrete insulin in response to both glucose and GLP-1/GIP receptor mediated potentiation. Insulin secretion measurement in EndoC-βH5 cells thus allow to measure dose responses to incretin analogs and to compare compound’s EC50s and maximum efficacies in a human beta cell environment, with endogenous receptor and signaling protein expression. In this study, we are now demonstrating that cAMP can also serve as a robust and rapid readout to measure GLP-1/GIP receptor signaling in these cells. We then compared EC50s and maximum efficacies obtained in glucose stimulated insulin secretion (GSIS) potentiation assay to those measured through cAMP production, this in order to characterize the predictivity of cAMP assays in the EndoC- βH5 human beta cell model. Overall, EndoC-βH5 cells, with several available readouts and endogenous expression of incretin receptor signaling machinery, is a powerful model to evaluate and compare the activity of incretin analogs on human beta cells and address the question of their mode of action in the case of pluri-receptor agonists.

Disclosure
M. Taurand: None. H. Olleik: Employee; Human Cell Design.

Funding
Pass Innovation Occitanie (France)
2103-LB: Cholinergic Innervation of the Islet Resident Macrophage
Authors: BOROWSKY NICHOLAS , CAICEDO ALEJANDRO 
Published: June 2024

Pancreatic islets are small endocrine organs containing beta cells and alpha cells that control glucose metabolism. These cells are supported by resident macrophages that tightly maintain tissue homeostasis. Islet resident macrophages respond to ATP secreted alongside insulin by increasing inflammatory cytokine expression. Thus, the normal physiology of insulin secretion has the potential to initiate an inflammatory response that needs to be limited. We propose that the cholinergic anti-inflammatory pathway provides a neural check on this inflammation. The cholinergic anti-inflammatory pathway reduces systemic inflammation in the case of sepsis and may play a role in other inflammatory conditions. However, there is no current understanding of the innervation of islet resident macrophages. To start addressing communication between cholinergic neurons and islet macrophages, we examined macrophages in the pancreas of transgenic mice expressing the green fluorescent protein GFP under the control of the choline acetyltransferase promoter. We found that islet macrophages were densely contacted by cholinergic terminals. We further studied functional responses of islet resident macrophages in living pancreas slices from transgenic mice expressing a fluorescent Ca2+ sensor (GCaMP6) under the control of the CSF1R promoter. Ca2+ responses to ATP in these macrophages were strongly inhibited in the presence of acetylcholine or nicotinic agonists. Oxotremorine, a selective muscarinic agonist, failed to dampen the macrophage response to ATP. Our findings showing that cholinergic nerves directly innervate islet resident macrophages and that selective nicotinic agonism modulates their responses to ATP suggesting that the cholinergic anti-inflammatory pathway provides neuroimmunomodulation in normal islet homeostasis. Further characterization of this pathway seems warranted as it could be harnessed to inhibit the chronic inflammatory conditions that trigger diabetes pathogenesis.

Disclosure
N.Borowsky: None. A.Caicedo: n/a.
216-OR: CREG1 Promotes Insulin Receptor Recycling to Enhance Insulin Signaling in Hepatocytes
Authors: JOO YECHAAN , QI YANMEI , RAHIMI SAUM A. , LEE LEONARD Y. , LI SHAOHUA 
Published: June 2024

CREG1 is an endolysosomal protein that promotes endocytic trafficking and lysosomal biogenesis (Autophagy, 2021). It is highly expressed in the liver. Recently, we demonstrated that the hepatocyte-specific deletion of the Creg1 gene in mice resulted in insulin resistance, elevated glycemia, and hyperinsulinemia. To investigate the role of CREG1 in hepatic insulin signaling and understand the underlying mechanisms, we stably transfected human and mouse hepatocytes with human CREG1 cDNA and mouse Creg1 shRNAs, respectively. CREG1 overexpression enhanced insulin signaling, as evidenced by phosphorylation of insulin receptor (InsR) and its downstream effectors Akt and GSK-3beta. Conversely, CREG1 depletion inhibited insulin-stimulated phosphorylation of these proteins. Immunofluorescence microscopy revealed a significant decrease in InsR expression on the hepatocyte surface, while the total InsR level remained unchanged in hepatocyte-specific Creg1 knockout mice. These novel findings suggest that loss of CREG1 may affect InsR trafficking. To test this hypothesis, we conducted live cell imaging of an InsR-GFP fusion protein and surface protein biotinylation. Pulse-chase experiments revealed that CREG1 overexpression increased insulin receptor recycling to the cell surface, while CREG1 depletion had the opposite effect. Furthermore, adenovirus-mediated InsR transfer to CREG1 knockdown hepatocytes not only restored InsR surface expression but also rescued insulin signaling. These results suggest that CREG1 regulates hepatic insulin signaling through its effects on insulin receptor recycling.

Disclosure
Y. Joo: None. Y. Qi: None. S.A. Rahimi: None. L.Y. Lee: Consultant; Abbott, Edwards Life Sciences. S. Li: None.
217-OR: Depalmitoylation Inhibition Improves Glucose Tolerance by Promoting Endothelial Insulin Transcytosis
Authors: ZHANG WEI , WEI XIAOCHAO , DONG GUIFANG , ADAK SANGEETA , WANG YIXUAN , SINGAMANENI SRIKANTH , SEMENKOVICH CLAY F. 
Published: June 2024

Mechanisms underlying insulin resistance are obscure, but insulin transport across endothelial cells is rate-limiting for insulin delivery to peripheral tissues. Protein palmitoylation regulates trafficking of endothelial organelles. Many acyltransferases mark proteins with palmitate, while acyl protein thioesterase 1 (APT1) is the dominant palmitate eraser, allowing protein cycling. We tested the hypothesis that APT1 affects glucose metabolism by mediating insulin access to target tissues. In two endothelium-specific knockout mouse models (Tie2-Cre and VE-cadherin-CreERT2), APT1 deficiency was associated with enhanced glucose clearance and increased insulin sensitivity on high-fat diet (HFD). HFD endothelial APT1 knockout mice had increased muscle phospho-Akt with similar body weight, body composition, and adipose inflammatory markers as compared to control mice. To assess insulin transcytosis across the endothelium, we used an immune-microneedle patch technique (employing 600-micron length microneedles coated with insulin antibody that penetrate the dermal layer of the skin) to measure insulin content of interstitial fluid (ISF). When the microneedle patch was applied to abdominal skin 10 min after administration of human insulin, ISF insulin in HFD endothelial knockout APT1 mice was ~60% higher (p=0.022) than in HFD control mice. When skin ISF was isolated by centrifugation, insulin determined by ELISA was ~50% higher (p=0.001) in HFD endothelial knockout APT1 mice than in HFD control mice. APT1 deficiency did not affect muscle endothelial morphology by electron microscopy. In human adipose microvascular endothelial cells, APT1 deficiency induced chemically or genetically promoted insulin transcytosis while APT1 overexpression decreased insulin transcytosis. These findings suggest that endothelial palmitoylation affects insulin transport from the blood to target tissues to mediate insulin sensitivity and glucose tolerance in mice.

Disclosure
W. Zhang: None. X. Wei: None. G. Dong: None. S. Adak: None. Y. Wang: None. S. Singamaneni: Stock/Shareholder; Auragent Bioscience LLC. C.F. Semenkovich: Consultant; Alnylam Pharmaceuticals, Inc.