bluuebunny/crossref_metadata_2025 huggingface dataset:
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.
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<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<0.001 and 1.56 [1.41-1.74]; p<0.001. Further, pro-BNP predicted MACE and cardiovascular death in patients with NAFLD (HRs 1.78 [1.54-2.07]; p<0.001 and 1.80 [1.53-2.12]; p<0.001, respectively) as well as in those who did not have NAFLD (HRs 1.47 [1.28-1.68]; p<0.001 and 1.51 [1.30-1.75]; p<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., <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< 0.01) and glucose SD, or glycemic variability (β= 0.30, p< 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 <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 >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<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<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<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] > 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 <5%), using DESeq2 and adjusting for batch and individual factors. This dramatic transcriptomic response was significantly enriched (FDR <5%) for response to hormone stimulus. Further, of 619,445 tested ATAC peaks, 7.3% (n=45,474) were significantly differentially accessible (FDR <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
291-OR: A Noninvasive Index for Significant Liver Fibrosis and Stratifying Risk of Cirrhosis in Patients with Type 2 Diabetes Mellitus in Primary Care
Authors: QI XIAOLONG , YOU DONGFANG , SHEN JIE , LIU CHUAN , CHEN YUPING , ZHENG MING-HUA , ZHOU XIQIAO , HUANG MINGXING , LIANG XUAN , DENG TIANXURUN , SUN CHAO , LIANG XIAO , WAN HENG , WANG YAN , ZHU YONGFEN , LV FANGFANG , ZHOU TAOLONG T. , LIU SHANGHAO , WANG XIAOMEI , LIU XING , HE RUILING , LI JIE , ZHAO YANG , WONG VINCENT W.
Published: June 2024

Introduction & Objective: To propose a non-invasive index for detecting patients with T2DM at risk of significant liver fibrosis and cirrhosis in primary care.

Methods: Five cohorts consist of 38,160 adults with T2DM were enrolled to develop and validate a non-invasive index for evaluating significant liver fibrosis and cirrhosis (Figure A).

Results: Risk factors including Aspartate transaminase and boDy mass inDex were fitted to develop a test, termed the ADD index (available at: https://chess.nuist.edu.cn/consult/gzconsult/add_index). Using transient elastography (TE) as the reference, the ADD index performed better than FIB-4 and NFS for significant fibrosis (Figure B). A cutoff of 0 of the ADD index was selected to stratify patients into low- and high-risk group that should receive TE. AUC of the ADD index followed by TE was significantly higher than that of FIB-4 and NFS followed by TE (Figure B). Notably, using the ADD index combined with TE resulted in 11% cost savings compared to use TE alone. Compared with patients in low-risk group, those in high-risk group had a 2.90-fold (95% CI:1.44-5.82, P < 0.01) risk for cirrhosis (Figure C).

Conclusions: The non-invasive ADD index can be applied to screen significant fibrosis and cirrhosis in patients with T2DM in primary care, and promoting those in high-risk group to further consult a hepatologist in secondary care.

Disclosure

X. Qi: None. D. You: None. J. Shen: None. C. Liu: None. Y. Chen: None. M. Zheng: None. X. Zhou: None. M. Huang: None. X. Liang: None. T. Deng: None. C. Sun: None. X. Liang: None. H. Wan: None. Y. Wang: None. Y. Zhu: None. F. Lv: None. T.T. Zhou: None. S. Liu: None. X. Wang: None. X. Liu: None. R. He: None. J. Li: None. Y. Zhao: None. V.W. Wong: Consultant; AbbVie Inc., Boehringer-Ingelheim, Echosens, Gilead Sciences, Inc. Research Support; Gilead Sciences, Inc. Consultant; Intercept Pharmaceuticals, Inc., Inventiva Pharma, Novo Nordisk, Pfizer Inc., Sagimet Biosciences.
292-OR: Coenzyme A Synthase Knockdown Alleviates Metabolic Dysfunction–Associated Steatohepatitis via Decreasing Cholesterol in Liver Lipid Droplets
Authors: SAKUMA IKKI , GASPAR RAFAEL C. , NASIRI ALI , KAHN MARIO , GUERRA MATEUS , YIMLAMAI DEAN , MURRAY SUE , PERELIS MARK , BARNES WILLIAM , VATNER DANIEL F. , PETERSEN KITT , SAMUEL VARMAN , SHULMAN GERALD I.
Published: June 2024

Metabolic dysfunction-associated steatohepatitis (MASH) is highly prevalent among patients with type 2 diabetes. Cholesterol is one of several candidates causing hepatic inflammation and fibrosis. We hypothesized that by interrupting the synthesis of cholesterol, we could prevent a choline-deficient L-amino acid-defined high-fat diet (CDAHFD) C57BL/6J MASH mouse model. We found that cholesterol in liver lipid droplets accumulated on day 1, MASH-associated macrophage markers (Gpnmb and Trem2) mRNA expression and plasma ALT increased on day 2, macrophage crown-like-structures emerged on day 3, and fibrosis markers (Col1a1, αSMA) mRNA expression increased on day 7 in the CDAHFD-treated mice. Filipin staining demonstrated free cholesterol accumulation in liver lipid droplets. We next examined whether a N-acetylgalactosamine-modified antisense oligonucleotide (ASO) against coenzyme A synthase (Coasy), which catalyzes a rate controlling step in hepatic cholesterol synthesis, would reduce cholesterol in liver lipid droplets and inflammation/fibrosis in this model. Coasy ASO treatment prevented accumulation of cholesterol in liver lipid droplets (Control=1.02 ± 0.04 mg/g tissue; Coasy=0.68 ± 0.08, P<0.001) and decreased plasma ALT and liver inflammation/fibrosis markers expression. To examine whether these effects of the Coasy ASO were mediated by reductions in liver lipid droplet cholesterol we examined whether adding 2% cholesterol to the diet would prevent the Coasy ASO protective effect. Cholesterol treatment induced accumulation of cholesterol in liver lipid droplets (Coasy=0.68 ± 0.08 mg/g tissue; Coasy with cholesterol =1.40 ± 0.11, P<0.001) and abrogated the protective effects of the Coasy ASO on liver inflammation and fibrosis.

Conclusions: These data demonstrate that cholesterol in liver lipid droplets is a key mediator of MASH and hepatic Coasy knockdown is a potential therapeutic approach.

Disclosure

I. Sakuma: None. R.C. Gaspar: None. A. Nasiri: None. M. Kahn: None. M. Guerra: None. D. Yimlamai: None. S. Murray: Employee; Ionis Pharmaceuticals. M. Perelis: Employee; Ionis Pharmaceuticals. W. Barnes: Employee; Ionis Pharmaceuticals. D.F. Vatner: None. K. Petersen: None. V. Samuel: None. G.I. Shulman: None.
299-OR: Discovery of Long-Acting Unimolecular Peptide Tetra-agonists Targeting GLP-1, GIP, Amylin, and Calcitonin Receptors for Enhanced Metabolic Benefits in Animal Models of Obesity
Authors: GHOSH SOUMITRA , VALDECANTOS PILAR , RADA PATRICIA , VALVERDE ANGELA M. , RONDINONE CRISTINA M.
Published: June 2024

Objective: The burgeoning prevalence of obesity and associated co-morbidities necessitates innovative approaches for safe and efficacious therapies. We describe the characterization of PTT-A, a novel long-acting peptide agonist for GLP-1, GIP, Amylin and Calcitonin Receptors, in rodent models and assessed its efficacy against the dual GIPR/GLP-1R agonist Tirzepatide.

Methods: In vitro assays used recombinant cell lines expressing individual receptors to assess GLP-1, GIP, Amylin and Calcitonin receptor agonism. Multiple metabolic endpoints were examined in rodents, including acute food intake and calcium regulation effects of PTT-A in lean rats, acute glucose-lowering effects in lean mice, and its chronic effects in diet-induced obesity (DIO) rats compared to Tirzepatide.

Results: PTT-A exhibited potent GIP, GIP, Amylin, and Calcitonin receptor agonism in cAMP assays, and induced decreases in blood glucose and calcium levels in acute studies in lean rodents. In lean rats, PTT-A dose-dependently reduced cumulative food intake. Chronic studies in DIO rats revealed significant reduction in cumulative food intake and body weight, driven by decreases in fat mass without loss of muscle mass, unlike that seen with Tirzepatide. PTT-A demonstrated robust efficacy for glucose and plasma lipid lowering, insulin sensitization and liver benefits, as indicated by lowered plasma insulin, alanine aminotransferase and liver triglycerides, while outperforming Tirzepatide at equivalent doses.

Conclusion: PTT-A is a novel tetra-receptor agonist, offering superior outcomes for weight loss, glycemic control, insulin sensitization, and liver health compared to Tirzepatide. These findings underscore the promise of such poly-pharmacological agents to tackle the complex challenges associated with obesity-related metabolic disorders.

Disclosure

S. Ghosh: Consultant; Abvance Therapeutics, Akero Therapeutics, Inc., Fractyl Health, Inc., Takeda Pharmaceutical Company Limited. P. Valdecantos: Research Support; Pep2Tango Therapeutics INC. P. Rada: Research Support; Pep2Tango Therapeutics INC. A.M. Valverde: Research Support; Pep2Tango Therapeutics INC. C.M. Rondinone: Board Member; Pep2tango Therapeutics Inc. Stock/Shareholder; Pep2tango Therapeutics Inc.
30-OR: Diabetes Rescue, Engagement, and Management (D-REM)—Results of a Pragmatic Clinical Trial of a Community Paramedicine Program to Improve Diabetes Care
Authors: DUCHARME-SMITH ALLISON , MCCOY ROZALINA G. , MEILANDER ANGELA K. , FISCHER KAREN , LIEDL CHAD , QUILLEN JAXON , JUNTUNEN MICHAEL B.
Published: June 2024

Introduction & Objective: People living with diabetes face multiple barriers to care and health, including gaps in access to medical care and diabetes self-management education/support (DSMES) and high costs of care. These challenges are particularly great for those living in rural areas. Community paramedics (CPs) are well positioned to deliver DSMES to patients with clinical and psychosocial complexities as part of an interdisciplinary clinical team. CPs are experienced paramedics with advanced training in the management of low acuity and chronic conditions who practice under the supervision of a physical medical director to provide a wide range of services tailored to each patient’s needs. While CP models have emerged across the U.S. and world to support transitional care, CPs have not been leveraged specifically for DSMES.

Methods: D-REM was a pilot pragmatic single-arm prospective trial of a community paramedic (CP) DSMES program that enrolled 70 adults with diabetes, hemoglobin A1c (HbA1c) ≥9%, and recent emergency department or hospital care for any cause. Participants received at-home CP care for approximately one month. Outcomes were ascertained from electronic health record review and postal surveys at 1- and 4-months.

Results: Enrolled participants were 61 (52.3-67.2) years old, 52.9% were men, and 50% lived in rural areas. D-REM resulted in 1.6% improvement in HbA1c, from 9.8% (8.9-10.6) at enrollment to 8.2% (7.1-9.3) at 4 months (p=0/02). It also improved engagement with primary care, endocrinology, and in-clinic DSMES. Participants were highly satisfied with their care.

Conclusion: CP is an emerging and scalable model of care delivery that can support diabetes management for patients experiencing clinical and structural barriers to care. Further research is needed to examine the effectiveness and implementation of CP programs in different settings and populations.

Disclosure

A. Ducharme-Smith: None. R.G. McCoy: Research Support; American Diabetes Association. Other Relationship; American Diabetes Association. Consultant; Wolters Kluwer Health. Research Support; National Institutes of Health, Patient-Centered Outcomes Research Institute. Consultant; Yale New Haven Health System. A.K. Meilander: None. K. Fischer: None. C. Liedl: None. J. Quillen: None. M.B. Juntunen: None.
31-OR: ROCKET Type 1 Diabetes—Launching Diabetes Self-Management Habits in Youth with New-Onset Type 1 Diabetes
Authors: ALARCON GUIDO , SONABEND RONA Y. , SCHWARTZ DAVID D. , LYONS SARAH , DESALVO DANIEL
Published: June 2024

Introduction: Performing the 6 T1D self-management habits (SMH) (Figure 1) has been associated with lower A1c and increased time in range (TIR) (Lee et al., JAMA Netw Open, 2021). We developed a remote patient monitoring program (RPM), called ROCKET T1D (Remote Outreach & Care for Kids’ Empowerment and Technology use in T1D), to optimize diabetes management in new onset T1D. We aimed to increase use of the 6 SMH by ≥ 2 habits from baseline in >80% of youth with newly diagnosed T1D.

Methods: At program launch, all youth with new onset T1D entered the ROCKET T1D RPM program within 2 weeks of diagnosis. After baseline assessment of the SMH, a CDCES conducted weekly virtual visits focused on each of the habits. After 3-month program completion, CDCES reassessed the SMH. CGM metrics were obtained in the first 2 weeks and again at program completion.

Results: Among 71 patients who completed the program, the mean cumulative habit score increased from 3.1 to 5.2, and 81% increased ≥ 2 habits from baseline. Mean TIR increased (56% to 64%, p=0.02) and mean GMI decreased (7.6% to 7.3%, p=0.07).

Conclusion: Remote support with tailored education following T1D diagnosis resulted in improved T1D management habits. CGM metrics were promising, but this could be due to other factors such as partial remission. Future work should evaluate whether early intensive education and technology initiation provides longterm glycemic benefit.

Disclosure

G. Alarcon: None. R.Y. Sonabend: None. D.D. Schwartz: None. S. Lyons: None. D. DeSalvo: Advisory Panel; Insulet Corporation. Consultant; Dexcom, Inc.

Funding

The Leona M. and Harry B. Helmsley Charitable Trust (2206-05307)
312-OR: OPAT Is Protective of Major Amputation following Hospitalization for Diabetic Foot Infection
Authors: TARRICONE ARTHUR , GEE ALLEN , REYES MARIO C. , LAVERY LAWRENCE A. , ROGERS LEE C.
Published: June 2024

Introduction & Objectives: Diabetic foot ulcers are a common complication of diabetes associated with high morbidity and mortality. It is estimated that the United States spends up to $13 billion USD each year on DFUs, with individual costs rising to $15,000 per year. Much of this expenditure has been attributed to lengthy hospital stays and inpatient care, with the need of prolonged antibiotic therapy. Outpatient antibiotic therapy (OPAT) is a service that provides intravenous antimicrobial medication in the outpatient setting and has been shown to reduce hospital stay and overall medical costs. This study aims to determine the benefit of OPAT therapy in a cohort with soft tissue and bone foot infections.

Methods: Prospective cohort study of patients hospitalized for diabetic foot infection. Patients were grouped by discharge status : OPAT program or standard of care. All patients were followed for a minimum of one year. Statistics were performed using Stata Be7. Continuous variables were reported as mean ± standard deviation and categorical variables were reported as their n(%). Odds ratios were written as O.R, followed by their associated confidence interval [CI]. Significance was determined as p≤0.05.

Results: A total of 246 patients were hospitalized with lower extremity infections in this cohort. OPAT was used in 93(38%) patients and 152(62%) with standard of care. The outcomes examined in this study included reinfection, binary wound healing, amputations, and major amputations. OPAT was found to be protective for major amputations ( OR= .21, 0.05-0.98, p=0.046). No difference was seen between reinfection ( OR=0.55, 0.28-1.09, p=0.085 ), binary wound healing ( OR=0.85, 0.50-1.45, p=0.54 or minor amputations ( OR=0.67, 0.22-1.95, p=0.45).

Conclusion: OPAT is protective against major amputation however does not reduce reinfection, improve binary wound healing, or reduce minor amputations.

Disclosure

A. Tarricone: None. A. Gee: None. M.C. Reyes: None. L.A. Lavery: None. L.C. Rogers: None.