bluuebunny/crossref_metadata_2025_split huggingface dataset:
1489-P: iPLA2gamma as a Mediator of Lipid-Induced Metabolic Inflammation and Insulin Resistance
Authors: LEE MYUNG-SHIK , KANG HYEREEN 
Published: June 2023

We have reported that bromenol lactone (BEL) improves metabolic profile. Here, we studied effect of BEL and its target iPLA2 in inflammasome which is a crucial in insulin resistance and metabolic inflammation. BEL inhibited inflammasome by LPS+palmitic acid (PA). BEL also ameliorated mitochondrial ROS and mitochondrial potential loss by LPS+PA, explaining reduced inflammasome. BEL administration in vivo also reduced metabolic inflammation and inflammasome. iPLA2gamma-KO but not iPLA2beta-KO macrophages showed reduced inflammasome by LPS+PA. Mitochondrial ROS and mitochondrial potential loss by LPS+PA were markedly reduced by iPLA2gamma KO. Cardiolipin content in macrophages was reduced by LPS+PA, which was recovered by BEL or myxothiazol, an inhibitor of mitochondrial complex bc1. Cardiolipin loss by LPS+PA was also ameliorated by iPLA2gamma KO. Cardiolipin liposome or expression of CRLS1, a gene for rate-limiting step of cardiolipin biosynthesis, reduced inflammasome by LPS+PA, while CRLS1 siRNA aggravated, suggesting that reduced cardiolipin content is critical in inflammasome by LPS+PA. iPLA2gamma-KO mice were protected from diet-induced insulin resistance, associated with reduced inflammasome and metabolic inflammation. These results suggest that initial reduction of cardiolipin content by PA-induced ROS probably due to removal of oxidized cardiolipin by iPLA2gamma might induce further mitochondrial ROS accumulation likely due to compromised mitochondrial complex activity, which would eventually lead to massive mitochondrial ROS production, marked loss of mitochondrial cardiolipin, profound mitochondrial dysfunction, and finally metabolic inflammation with lipid-induced insulin resistance, through a feed-forward mechanism. Replenishment of cardiolipin could a novel modality for treatment of diet-induced insulin resistance through recovery of mitochondrial function and amelioration of lipid-mediated inflammasome or metabolic inflammation.

Disclosure
M.Lee: None. H.Kang: None.

Funding
National Research Foundation of Korea (2019R1A2C3002924)
1494-P: ILV-001, a Highly Promising Therapeutic Agent for Islet Transplantation
Authors: LANGLOIS ALLAN , GRABARZ ANASTASIA , PINGET MICHEL , BOUZAKRI KARIM 
Published: June 2023

Introduction: Our team has shown a bimodal effect on β-cells of muscle-secreted myokines from normal or insulin-resistant human skeletal muscle. We identified ILV-001, a component of the muscle secretome regulated by muscle fiber type and physical exercise, moreover naturally present in the extracellular matrix of the islet niche. Thus, we aimed at investigating the therapeutic potential of ILV-001 in protecting β-cell function and survival during islet transplantation.
Methods: In vitro: Rat islets were incubated 24h ± ILV-001 (1µg/mL). Islet survival was assessed by TUNEL assay ± cytomix. Islet function was evaluated using GSIS test. In vivo: 10000 IEQ/kg ± ILV-001 were injected intraportaly in diabetic syngeneic rats. For 3 months body weight gain, glycaemia, c-peptidemia and graft function (IPGTT) were followed. Inflammation was evaluated titrating plasmatic α2-macroglobulin. Histology was performed on grafted islets.
Results: In vitro: ILV-001 improves islet function and survival and potentiates insulin secretion after GSIS test. In vivo: ILV-001 improves graft function in terms of glycaemia (p&lt;0.001) and c-peptidaemia (p&lt;0.01) and decreases exogenous insulin intake requirements. ILV-001 reduced inflammation 24h post graft with a lower plasmatic α2-macroglobulin versus control with respectively 459±70 versus 903±191 µg/mL (p&lt;0.05). Finally, histology showed a preservation of insulin positive cells with ILV-001 versus control at the end of study (69±3% versus 39±3% of β-cells/islet, p&lt; 0.001).
Conclusion: ILV-001 is a highly promising candidate for the achievement of a functional cure in T1D thanks to potent β-cell preservation and obtaining long term graft maintenance following islet transplant.

Disclosure
A.Langlois: None. A.Grabarz: Stock/Shareholder; Ilonov. M.Pinget: Consultant; Novo Nordisk, Stock/Shareholder; ILONOV. K.Bouzakri: Consultant; Adocia, Stock/Shareholder; ILONOV.
1499-P: Adipocyte Development from Mesenchymal Progenitor Cells Differ between Subjects with Normal Glucose Tolerance or Type 2 Diabetes
Authors: CORVERA SILVIA , DESOUZA TIFFANY A. , ZIEGLER RACHEL N. , ROSANO NINA , THOMPSON MICHAEL J. 
Published: June 2023

Subcutaneous adipose tissue protects from T2D by sequestering lipids and preventing lipotoxicity and insulin resistance. Adipose depots are maintained throughout the lifetime by the formation of new adipocytes from mesenchymal progenitor cells that reside within the tissue. In previous studies, we have found that mesenchymal progenitor cells from human adipose tissue can be expanded with little loss of multipotency by culturing tissue explants in 3-dimensional hydrogels under pro-angiogenic conditions. Mesenchymal progenitor cells obtained in this manner can be induced to differentiate into multiple adipocyte subtypes, including beige adipocytes characterized by their induction of UCP1 upon cAMP elevation. Using this approach, we explored whether mesenchymal progenitor cells from the subcutaneous abdominal (ABD) and gluteal (GLU) depots of subjects with normal glucose tolerance (NGT) or T2D would differ in their capacity to differentiate into adipocyte subtypes. Results from n=12 subjects (6 with NGT and 6 with T2D) revealed no significant differences between depots or glycemic states in the number of mesenchymal progenitor cells that can be expanded, in their proliferative rate upon subsequent passaging, or in their capacity to differentiate into either white or beige adipocytes. However, we found that cells from either the ABD or GLUT depot from NGT subjects displayed substantial spontaneous adipocyte differentiation even without adipogenic induction, but cells from subjects with T2D did not. This difference was highly significant, and potentially reflects a diminished capacity of mesenchymal progenitor cells to maintain subcutaneous adipose depots in-vivo in subjects with T2D. Further transcriptomic and epigenomic analyses of expanded progenitor cells, currently underway, has the potential to reveal cellular mechanisms of impaired spontaneous adipocyte differentiation in subjects with Type 2 Diabetes.

Disclosure
S.Corvera: None. T.A.Desouza: None. R.N.Ziegler: None. N.Rosano: None. M.J.Thompson: None.
150-LB: Evaluation of a Diabetes Screening Clinical Decision Support Tool
Authors: TSENG EVA , MATHIOUDAKIS NESTORAS , MARUTHUR NISA M. , YEH HSIN-CHIEH , STEIN ARIELLA 
Published: June 2023

As part of a health system diabetes prevention initiative, we implemented and evaluated if an EHR clinical decision support tool improves diabetes screening rates. The EHR clinical decision support tool, implemented in 7/2021, alerts clinicians when a patient is due for diabetes screening based on USPSTF 2021 guidelines (BMI ≥25 kg/m² and age 35-70 years). The primary outcome was the monthly screening lab order rate, % patients eligible for screening who received a lab order during their primary care visit in that month. The secondary outcome was the monthly screening lab completion rate, % patients eligible for screening who completed a screening lab test during their primary care visit in that month. We used logistic regression to evaluate changes in outcomes from baseline to post-implementation periods. In a tertiary health system, 16,431 eligible primary care patients were seen between 7/2020-12/2022. At baseline (7/2020-7/14/21), screening lab order and completion rates were 65% and 58%, respectively. Rates increased significantly in the post-implementation period (1/2022-12/2022) compared to baseline (lab orders: OR 1.7, p&lt;0.001, completion: OR 1.8, p&lt;0.001). Our study demonstrates an EHR clinical decision support tool, in the context of a system-wide initiative, can successfully improve diabetes screening rates. These findings support other healthcare organizations to explore clinical support tools for diabetes screening.

Disclosure
E. Tseng: None. N. Mathioudakis: None. N. M. Maruthur: Other Relationship; Johns Hopkins HealthCare Solutions. H. Yeh: None. A. Stein: None.

Funding
National Institutes of Health (K23DK118205)
1502-P: Adipose Tissue Protein Response and Insulin Sensitivity in Obesity
Authors: LYTLE KELLI , JENSEN MICHAEL D. 
Published: June 2023

Dysregulation of adipose tissue (AT) lipolysis, a hallmark of obesity and type 2 diabetes, results in elevated postabsorptive and postprandial plasma free fatty acid (FFA) concentrations. This is thought to be due to adipose insulin resistance with regards to anti-lipolysis effects. Alterations in intracellular adipocyte pathways and/or proteins that mediate this phenomenon in humans are poorly understood. To test the hypothesis that impaired insulin signaling is associated with adipose insulin resistance measured whole body insulin sensitivity (the insulin concentration that suppresses palmitate flux by 50% (IC50 FFA)) in 12 lean, 7 lower body obese and 15 upper body obese (n=15) volunteers using a single step hyperinsulinemic-euglycemic clamp. We also measured adipose pAkt/Akt (a measure of insulin signaling) and hormone sensitive lipase (pHSL/HSL) (a measure of lipolysis protein regulation) in biopsies of abdominal and thigh adipose obtained before and during the insulin clamp were measured. Contrary to our hypothesis, there was a positive relationship between IC50 FFA and insulin clamp fold change of pAkt/Akt in thigh samples (ρ = 0.38, p &lt;0.05) and no relationship for abdominal adipose samples. There was good correlation between abdominal and thigh adipose samples in the fractional change in pAkt/Akt in response to insulin clamp (ρ= 0.49, p &lt; 0.01). IC50 FFA was correlated to fractional decrease in HSL phosphorylation in both abdominal (r = 0.36, p&lt; 0.05) and thigh (r = 0.41, p &lt;0.05) Surprisingly, fold changes of pAKT/AKT in response to insulin clamp were positively correlated to fold change of pHSL/HSL in abdominal adipose tissue (ρ= 0.41, p &lt; 0.05) but not thigh samples. In conclusion we found that insulin resistance in adipose is not related to canonical insulin signaling responses but is related to lipase responses.

Disclosure
K.Lytle: None. M.D.Jensen: Other Relationship; Novo Nordisk, Elsevier.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (DK40484)
1503-P: Insulin Regulates Expression and Secretion of Exosomal MicroRNAs in Adipocytes
Authors: LINO MARSEL , PAN HUI , DREYFUSS JONATHAN , RONALD KAHN C. 
Published: June 2023

Transfer of regulatory molecules, such as microRNAs (miRNAs), via small extracellular vesicles (sEVs) called exosomes represents a newly recognized mode of inter-organ communication that provides an important mechanism of metabolic regulation. We have previously shown that adipose tissue is a major contributor to the pool of circulating exosomal miRNAs and that these adipose-derived exosomal miRNAs can be taken up by target tissues and suppress gene expression. We have also shown that biogenesis, secretion, and exosomal cargo sorting is a highly regulated process and that miRNAs contain 4-8 nucleotide motifs which contribute to exosomal secretion. Several exosomal miRNAs have been linked to obesity and diabetes, yet their regulation by physiological stimuli such as insulin remains unknown. To identify the role of insulin in regulation of adipose miRNAs and their secretion in exosomes, we performed small RNA-seq on exosomes isolated from differentiated 3T3-L1 cells with and without insulin stimulation. We find that out of the 460 miRNAs detected, 39 were up- and 75 were down-regulated in exosomes in response to insulin, whereas only 14 were up- and 39 were down-regulated in cells. In some cases, exosomal secretion correlated with cellular expression of the miRNA. More interestingly, 103 of the 114 miRNAs (90.3%) that were regulated in exosomes had no significant change at the cellular level or even a change in the opposite direction, demonstrating insulin’s ability to regulate miRNA exosomal secretion independent of cellular level. These insulin-regulated exosomal miRNAs included members of the let-7 family, which have known functions in adipogenesis, as well miRNAs predicted to target mRNAs of molecules involved in fatty acid biosynthesis and insulin signaling. Thus, insulin can independently regulate miRNA expression and secretion in adipocyte-derived exosomes. These can serve as adipokines and create a novel mechanism for regulation of cell-cell and inter-organ communication.

Disclosure
M.Lino: None. H.Pan: None. J.Dreyfuss: None. C.Kahn: None.

Funding
National Institutes of Health (5R01DK082659)
1505-P: CD44 Knockdown in Human Adipocytes—Effect on Adipocyte Proliferation and Differentiation
Authors: CHEN HENRY , SAINI NATESH R. , AYHAN EKREM M. , MCLAUGHLIN TRACEY 
Published: June 2023

Introduction: eGWAS implicates the immune-cell receptor CD44 as the top candidate gene in the molecular pathogenesis of T2D1. We have shown that in human adipose tissue, CD44 expression is associated with inflammatory gene expression, M1 macrophage phenotype, and systemic insulin resistance2. Here we sought to determine whether CD44 directly impairs adipocyte function via siRNA knockdown in human preadipocytes with quantification of proliferation, differentiation, and gene expression.
Methods: Abdominal subcutaneous adipose tissue was harvested from five healthy nondiabetic obese individuals during bariatric surgery. Preadipocytes were isolated via collagenase digestion and seeded at 1×10^4 cells/well and treated with siRNA-CD44 or mock condition (control) using siRNA Maxi Transfection Reagents (Invitrogen). Cell proliferation was assayed by MTS (Promega) on Days 1, 2, 3 post transfection. Differentiation was induced at confluence (as described3), and cells were harvested at days 3, 6, 12 after confluence for gene expression and at day 12 for oil red O staining (total lipid uptake) and immunofluorescence microscopy (% differentiation = bodipy/dapi*100).
Results: CD44 knockdown decreased preadipocyte proliferation and increased adipocyte differentiation and lipid uptake.
Conclusion: CD44 may contribute to human insulin resistance by impairing adipocyte differentiation and fat storage.

Disclosure
H.Chen: None. N.R.Saini: None. E.M.Ayhan: None. T.Mclaughlin: Board Member; January, Inc., Research Support; Merck & Co., Inc., Stock/Shareholder; Eiger Bio.

Funding
National Institutes of Health (1R21AI159024-01A1)
1507-P: Carnosic Acid–Induced Browning in 3T3-L1 Adipocytes Is Dependent on AMPK Activation
Authors: VLAVCHESKI FILIP , TSAKIRIDIS NICOLE , GIACCA ADRIA , TSIANI EVANGELIA 
Published: June 2023

Background: Increased adiposity has long been associated with insulin resistance and adipose tissue plays a crucial role in regulating metabolic homeostasis, as its dysfunction in obesity leads to insulin resistance and type 2 diabetes (T2D). The primary function of white adipose tissue (WAT) is to store energy as lipids while brown adipose tissue (BAT) regulates thermogenesis by dissipating energy in a form of heat. The process of browning involves transdifferention of WAT into brown-like or beige adipocytes, which exhibit the same functional properties as BAT. Browning of WAT is an attractive approach against obesity and insulin resistance. In addition, evidence indicate that activation of the energy sensor AMP-activated protein kinase (AMPK) could counteract insulin resistance. The aim of this study is to examine if carnosic acid (CA), induces browning via activation of AMPK in 3T3-L1 white adipocytes.
Material and methods: Cell morphology, lipid accumulation, mitochondrial density, and browning markers such as uncoupling protein-1 (UCP-1) and peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) levels were assessed using Cytation5 microscope, Oil-O-Red (ORO), MitoTracker Red, immunoblotting and immunocytochemistry.
Results: Microscopic investigation revealed that CA reduced the size and number of lipid droplets in the cells, giving a rise to a brown-like phenotype. The reduction of lipid accumulation was also confirmed by ORO stain. Next, CA increased mitochondrial density and protein expression of UCP-1 and PGC-1-α in 3T3-L1 adipocytes. Most importantly, pre-treatment with compound C, prevented the CA-induced reduction in lipid accumulation, and mitochondrial density and blocked the CA-induced increase in UCP-1 and PGC-1 expression.
Conclusion: This study demonstrates that CA has a pronounced potential in stimulating browning of adipocytes via AMPK-dependent mechanism. Future in vivo studies are required to fully examine the effects of CA.

Disclosure
F.Vlavcheski: None. N.Tsakiridis: None. A.Giacca: None. E.Tsiani: None.

Funding
Natural Science and Engineering Research Council of Canada (RGPIN-2018-06666)
1509-P: Global Metabolomic Profiling and the Pathobiology of Prediabetes and Type 2 Diabetes
Authors: DUBEY DHRUV , DUTTA TUMPA , NUNEZ LOPEZ YURY O. , GARDELL STEPHEN J. , PRATLEY RICHARD E. 
Published: June 2023

Introduction: Metabolomics has provided valuable insights into metabolic alterations associated with diabetes. However, prior studies profiled only a small subset of metabolites that was possible with the contemporaneous technology. We aimed to reassess and extend the current knowledge by implementing comprehensive LC-MS/MS metabolomic analyses.
Methods: Untargeted metabolomic analysis was performed using serum samples from people with normal glucose tolerance, prediabetes, and type 2 diabetes (n=30, balanced ORIGINS cohort subset, NCT02226640), at baseline and 2 hours (2h) after an oral glucose tolerance test (OGTT). Differential expression (MetaboAnalyst) and coexpression network (WGCNA) analyses were implemented. Random Forest classification was used to further assess variable importance.
Results: A total of 697 known metabolites were detected in more than 80% of the samples. This represents a significant increase in the number of detected circulating metabolites compared to previous studies in diabetes. WGCNA identified modules of correlated metabolites that associated with glucose tolerance at baseline and 2h. The top module for each timepoint was enriched in metabolites that significantly changed in response to the glucose challenge and highlighted an underappreciated role of glyoxylate and dicarboxylate metabolism in early stages of diabetes evolution. Elevation of hexoses and bile acids, and downregulation of amino acids, lipids, and free fatty acids were demonstrated after the glucose challenge. Random forest classification further unveiled the biomarker potential of baseline levels of aminoadipic and methyluric acids.
Conclusions: Comprehensive profiling of serum metabolites in humans underscored the role of carbohydrate metabolism, the citric acid and urea cycles, suppression of lipolysis, ketogenesis, and proteolysis in response to a glucose challenge. Potential biomarkers of early diabetes development and progression were identified.

Disclosure
D.Dubey: None. T.Dutta: None. Y.O.Nunez lopez: None. S.J.Gardell: None. R.E.Pratley: Other Relationship; Bayer Inc., Corcept Therapeutics, Dexcom, Inc., Gasherbrum Bio, Inc., Hanmi Pharm. Co., Ltd., Hengrui (USA) Ltd., Merck Sharp & Dohme Corp., Novo Nordisk, Pfizer Inc., Rivus Pharmaceuticals Inc., Sanofi, Scohia Pharma Inc., Sun Pharmaceutical Industries Ltd.

Funding
AdventHealth Translational Research Institute
151-OR: Social Determinants of Health Predict Diabetic Foot Infection Development
Authors: SCHMIDT BRIAN M. , BURANT AARON , BAKER NICOLE , KATONA AIMEE , MARTIN CATHERINE , HOLMES CRYSTAL M. , BUSUI RODICA 
Published: June 2023

The objective was to identify social determinants of health (SDOH) associated with development of diabetic foot infection using the CDC Healthy People Framework at a tertiary care center. Targeted interrogation of structured and unstructured data using novel electronic medical record search engines was used to analyze patient encounters in individuals with diabetes and a concomitant diabetic foot complication during a twenty-year period (2001-2021). Among ~ 200 million encounters from people with diabetic foot complications, 8351 developed diabetic foot ulcers (DFU), and 1671 suffered a DFU infection. The risk of infection development in people with a DFU was assessed with the neighborhood disadvantaged index score from over 200 million separate patient encounters. The neighborhood disadvantaged index based on available geolocation data, provides a unitless value to describe the relative socioeconomic background of an individual as determined by the proportion (%) of non-Hispanic black, proportion (%) of female headed families with children, and the proportion of households (%) with public assistance income or food stamps in a defined area. The average disadvantaged index score for the DFU population was 0.06 and was comparable to the state of Michigan's average disadvantaged index of 0.05. In those persons who developed DFU infection, the index score averaged 0.13, 2.2x higher than the DFU population (p&lt;0.05). Additionally, divorced individuals represented 2% of all individuals with DFU but accounted for 5% of those who developed DFU infection. This was disproportionate to other marriage statuses. These data strongly support the need to incorporate SDOH into updated DFU standard of care guidelines. Moreover, we clearly demonstrate that SDOH, more specifically geolocation data, can be used to identify individuals who are more likely to develop DFU infection. Thus, it is essential that SDOH are included in patient risk stratification to prevent severe diabetic foot complications.

Disclosure
B.M.Schmidt: None. A.Burant: None. N.Baker: None. A.Katona: None. C.Martin: Advisory Panel; Nevro Corp. C.M.Holmes: None. R.Busui: Board Member; American Diabetes Association, Consultant; Averitas Pharma, Inc., Lexicon Pharmaceuticals, Inc., Nevro Corp., Novo Nordisk, Roche Diagnostics, Procter & Gamble, Research Support; Novo Nordisk, Medtronic, National Institutes of Health.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (1K23DK131261-01A1, 5U01DK119083-05)