bluuebunny/crossref_metadata_2025 huggingface dataset:
1357-P: Diabetic Hyperglycemia Impairs Intestinal Barrier Function in the Setting of Diet-Induced Obesity
Authors: FRANCIS KENDRA L. , ALONGE KIMBERLY M. , HU SHANNON J.W. , PHAN BAO ANH N. , DOAN TAMMY P. , KRUTZSCH CODY A. , MAAT ZENOURA , SCARLETT JARRAD M. , SCHWARTZ MICHAEL W.
Published: May 2022

Proper function of the intestinal barrier plays a crucial role to maintain normal health by selectively absorbing nutrients and keeping out pathogens. Increased gut permeability is a feature of several diseases, including metabolic syndrome, but whether it is a cause or a consequence is unknown. We therefore examined the impact on intestinal barrier function of streptozotocin (STZ) -induced diabetes in C57BL/6J mice that were fed either standard chow or high-fat high-sucrose (HFHS) diet for 13wk. Moderate hyperglycemia was induced by low-dose i.p. STZ injection and 12wk later, intestinal tissues were collected and processed for quantification of colonic mucin and tight junction proteins. In both HFHS- and chow-fed mice, STZ treatment resulted in significantly elevated blood glucose levels compared to vehicle-treated controls (HFHS blood glucose 249.1 mg/dL vs. 157.5 mg/dL, p=0.0008; chow 330.2 mg/dL vs. 139.7mg/dL, p=0.0005) . In HFHS-fed mice, STZ hyperglycemia was associated with reductions of both the colonic mucin layer, as measured by Alcian Blue (AB) staining (percent area 52.6 vs. 62.4, p<0.0001) , and the tight junction protein E-cadherin (percent area 34.7 vs. 55.9, p=0.0029) . In non-obese chow-fed mice, however, STZ-induced hyperglycemia had no significant impact on AB mucin staining or E-cadherin protein abundance. Similarly, levels of colonic mucins and E-cadherin were inversely correlated with blood glucose levels in HFHS-fed mice (AB R2=0.43, p=0.0116; E-cadherin R2=0.30, p=0.0439) , but not in chow-fed mice. We conclude that while the combination of hyperglycemia and diet-induced-obesity is sufficient to impair intestinal barrier function, neither obesity nor diabetes in isolation is sufficient to induce this effect. Our additional finding that the magnitude of colonic dysfunction is correlated with the degree of hyperglycemia implicates excess glucose as a potential driver of obesity-associated impairment of intestinal barrier function.

Disclosure

K.L.Francis: None. K.M.Alonge: None. S.J.W.Hu: None. B.N.Phan: None. T.P.Doan: None. C.A.Krutzsch: None. Z.Maat: None. J.M.Scarlett: None. M.W.Schwartz: None.

Funding

National Institutes of Health T32 DK007742
1359-P: Effect of Fat Content on Postprandial Gastric Retention and Glucose Absorption in Subjects with Type 1 Diabetes during Daily Life Conditions: Assessment through a Computational Model
Authors: FAGGIONATO EDOARDO , SCHIAVON MICHELE , BUCKINGHAM BRUCE A. , EKHLASPOUR LAYA , MAN CHIARA DALLA
Published: May 2022

Understanding the impact of dietary nutrients on postprandial gastric retention (GR) and glucose absorption (GA) is crucial for optimizing both open- and closed-loop therapy in type 1 diabetes (T1D) . Here, we aim to investigate the effect of fat (F) content of a meal on both GR and GA in real life conditions, using a recently developed minimally invasive oral minimal model (MI-OMM) .

Data was collected during an outpatient 5-days clinical trial involving 1patients (age = 3-61 y; M/F = 60/50) wearing a sensor augmented insulin pump. At each meal, the medical staff recorded carbohydrate (C) and F intake. A total of 159 meals were analyzed that included >75% sensor data, C and F contents, no exercise sessions, or other meals/snacks within 2h before and 3h after the meal. Meals were grouped based on F content: high fat (HF, F≥30%) , and Low Fat (LF, F≤15%) .

The model well fitted glucose sensor data and provided estimated GR and GA curves. Fig. 1 shows that model-derived GR was significantly lower from 0.5h onward, and GA was significantly faster from 0.5h to 2.3h after a LF meal compared to a HF.

These findings proved that MI-OMM can be used to estimate GR and GA, using sensor data, in real life meals with different F content and, therefore, it is potentially applicable to optimize insulin therapy in T1D.

Disclosure

E.Faggionato: None. M.Schiavon: None. B.A.Buckingham: Advisory Panel; Arecor, Lilly Diabetes, Medtronic, Other Relationship; Insulet Corporation, Research Support; Insulet Corporation, Lilly Diabetes, Medtronic. L.Ekhlaspour: Consultant; Tandem Diabetes Care, Inc., Ypsomed AG. C.Dalla man: Research Support; Becton, Dickinson and Company, Sanofi-Aventis Deutschland GmbH.

Funding

This work was supported by MIUR (Italian Minister for Education) under the initiative “Departments of Excellence” (Law 232/2016) . Laya Ekhlaspour is funded by NIH (National Institutes of Health) 1K23DK121942.
136-OR: Polygenic Scores Improve the Predictive Accuracy of Type 1 Diabetes Classification Algorithms in a Large Biobank Cohort
Authors: DEUTSCH AARON J. , MAJARIAN TIM , MERCADER JOSEP M. , FLOREZ JOSE C. , UDLER MIRIAM
Published: May 2022

Introduction: Many biobanks use machine learning algorithms to predict patient phenotypes such as type 1 diabetes (T1D) . It is not known how these algorithms perform in patients with different ancestral backgrounds or whether incorporating genetic data improves phenotypic classification.

Methods: We examined 40,843 patients in the Mass General Brigham (MGB) Biobank, including 34,870 white patients and 4,872 non-white patients. The MGB machine learning algorithm predicted 242 patients to have T1D. After applying a gold standard diagnosis based on chart review, we assessed accuracy of the MGB algorithm in different self-reported races and genetic ancestries. We divided the dataset into separate training and validation cohorts, and we constructed a logistic regression model using the machine learning algorithm and previously published T1D polygenic scores to classify patients as T1D cases or controls.

Results: The MGB machine learning algorithm for T1D had a positive predictive value of 77.8% for white patients, but only 62.2% for non-white patients (p = 0.04) . Among patients with predominantly European genetic ancestry, adding a polygenic score derived from European populations to the MGB classification algorithm improved the area under the receiver operating curve (AUC) from 0.928 to 0.974 (p < 0.05) . Among patients whose predominant genetic ancestry was not European, adding a polygenic score derived from African populations improved the AUC from 0.889 to 0.920 (p > 0.05) . In both cases, the improvement in AUC was less substantial when using non-ancestry-matched polygenic scores.

Conclusions: Automated prediction tools for T1D are imperfect, and performance may differ by patient race. Incorporating ancestry-specific polygenic scores into phenotyping algorithms can improve diagnostic accuracy. This may help reduce healthcare and research disparities by refining the automated classification of T1D in all racial and ancestral groups.

Disclosure

A.J.Deutsch: None. T.Majarian: Employee; Vertex Pharmaceuticals Incorporated. J.M.Mercader: None. J.C.Florez: Consultant; AstraZeneca, Goldfinch Bio, Inc., Other Relationship; AstraZeneca, Merck & Co., Inc., Novo Nordisk. M.Udler: None.

Funding

National Institutes of Health (K23DK114551, T32DK007028)
1361-P: D-Xylose Test as a Biomarker for Glucose Intestinal Absorption in Humans and Minipigs
Authors: GOUTCHTAT REBECCA , MARCINIAK CAMILLE , CAIAZZO ROBERT , VERKINDT HELENE , QUENON AUDREY , RABIER THIBAUD , LAPIERE SARAH , RAVERDY VIOLETA , HUBERT THOMAS , PATTOU FRANCOIS
Published: May 2022

Background and Aim: Increasing evidences suggest that intestinal glucose absorption (IGA) plays a distinct role in pathophysiology of type 2 diabetes (T2D) . However, state of the art IGA measurement with multiple tracers techniques are demanding and . We propose D-xylose, a non metabolized pentose, as a proxy to quantify IGA levels to study IGA involvement in T2D.

Materials and Methods: First (cross sectional studies) , mixed meal tests including D-xylose (MMT) were performed in 165 patients (lean normoglycemic (NG) ; obese NG, glucose intolerant (IGT) or T2D) and 74 healthy adult minipigs. Second (longitudinal studies) , MMT were performed in minipigs before and after 70% intestinal resection (n = 5) , 70% pancreatectomy (n = 15) , and high fat diet for 2 months (HFD) (n = 4) to quantify D-xylose changes after each intervention.

Results: 1h serum D-xylose varied with glucose tolerance status (p<0.005) and was higher in T2D and IGT patients than in obese and lean NG patients In multivariable analysis, 1h postprandial glucose was correlated with D-xylose (r= 0.63; p<0.0001) , independently of insulin secretion (insulinogenic index) and insulin sensitivity (Matsuda index) . In minipigs, D-xylose was also independently correlated with post prandial glucose (r=0.53; p<0.05) . D-xylose appearance was reduced after intestinal resection (p<0.05) , but remained unchanged after pancreatectomy and HFD.

Conclusion and Perspectives: Post prandial glucose was independently correlated with D-xylose in humans and minipigs. Interventional studies in minipigs provided direct evidence that D-xylose is associated with IGA, independently of insulin secretion and sensitivity.

Disclosure

R.Goutchtat: None. F.Pattou: None. C.Marciniak: None. R.Caiazzo: Consultant; Ethicon, Inc., Medtronic. H.Verkindt: None. A.Quenon: None. T.Rabier: None. S.Lapiere: None. V.Raverdy: None. T.Hubert: None.

Funding

ANR
1369-P: The Differential Effects of Estrogen on Determinants of Skeletal Muscle Bioenergetics with a High-Fat, High-Sucrose Diet in Rats
Authors: HULETT NICHOLAS A. , HULL SARA , KNAUB LESLIE , POTT GREGORY , MILLER BENJAMIN F. , REUSCH JANE , SCALZO REBECCA L.
Published: May 2022

Estrogen is a powerful, beneficial regulator of skeletal muscle mitochondria. Premenopausal women with type 2 diabetes (T2D) have relatively greater complications associated with their T2D compared with the effect of the disease in men. We hypothesize that this impact of T2D in women is in part due to the impairment of estrogen-mediated regulation of skeletal muscle mitochondrial content and function. To explore the interaction between estrogen, diabetes and mitochondrial biogenesis in a rat model, 4-week-old, female Wistar Rats were fed a chow or a high fat/high sugar diet (HFHS) . At 8 weeks all mice underwent an ovariectomy (OVX) and randomized to estradiol (E2) replacement or none. One week prior to sacrifice, deuterium oxide was administered to determine skeletal muscle mitochondrial biogenesis. OVX +/- E2, animals were sacrificed 6 weeks post-randomization. Endpoints: body weight, gonadal fat mass, blood glucose and insulin levels, and gastrocnemius (mitochondrial biogenesis and function) . OVX+E2 rats had lower body mass compared with OVX (P = 0.0028) . HFHS fed rats had greater relative fat mass independent of E2 status (P = 0.0007) , and the OVX+E2 fed the HFHS diet were hyperglycemic (P = 0.00for the diet x E2 interaction) . State 3 mitochondrial respiration with carbohydrate-linked substrates was elevated in chow fed animals with E2, and this response was absent in HFHS animals (P = 0.0138 for the diet x E2 interaction) . Contrary to our hypothesis, there was no effect of diet or E2 on mitochondrial biogenesis measured with deuterium oxide (P > 0.4) . These data support the hypothesis that, in the context of a HFHS diet, the beneficial effect of E2 on skeletal muscle mitochondrial function is impaired. However, mitochondrial biogenesis does not appear to mediate this interaction of diet and E2. Future studies will focus on the E2-mediated regulation of mitochondrial quality to determine their role in the impairment of mitochondrial function.

Disclosure

N.A.Hulett: None. S.Hull: None. L.Knaub: None. G.Pott: None. B.F.Miller: None. J.Reusch: Advisory Panel; Medtronic. R.L.Scalzo: None.

Funding

Veterans Affairs (BX004533)
1376-P: Comparison of a-Cell Response to Intravenous or Oral Glucose in Nondiabetic Humans
Authors: LAURENTI MARCELLO C. , MANDUCA ARMANDO , COBELLI CLAUDIO , MAN CHIARA DALLA , VELLA ADRIAN
Published: May 2022

Type 2 Diabetes (T2DM) and prediabetes are characterized by β-cell and α-cell dysfunction which can occur independently. Defects in α-cell response to rising glucose can be evaluated with an oral or intravenous challenge: the former reflects normal physiology, the latter is unaffected by incretins or gut motility. We compared Glucagon Secretion Rate (GSR) in response to rising glucose during a Graded Glucose Infusion (GGI) and an Oral Glucose Tolerance Test (OGTT) . We studied 39 non-diabetic, weight-stable subjects (53 ± 2 yrs, 29 ± 1 Kg/M2) , with different fasting and glucose tolerance status. All subjects underwent a 75g OGTT and a GGI, with glucose infusion doubling every hour for 4 hours. GSR was calculated by deconvolution from plasma concentrations of glucagon on both study days (GSRGGI and GSROGTT) . The suppression of GSRGGI could be described with a single-exponential relationship between GSR and glucose concentrations. The slope of the log-transformed secretion line allows estimation of the change in glucose necessary to suppress GSR by 50% (G50) . This index of suppressibility is elevated in people with impaired fasting glucose compared to normal fasting glucose (2.7 ± 0.2 vs. 3.8 ± 0.4 mmol/L, p=0.01) . Intriguingly, GSROGTT, exhibited a delayed response to rising glucose (compared to GSRGGI) , during the early part of the OGTT. However, the relationship of GSROGTT to glucose did not differ from that of GSRGGI during the fasting state and at the time of peak postprandial suppression. This allowed the calculation of G50 during the OGTT, which was similar to that observed during the GGI (R2=0.66) and correlated with weight, age and fasting glucose (R2=0.49) . These data demonstrate the ability of a novel index (G50) , to reproducibly quantify α-cell suppression by oral or intravenous glucose in non-diabetic subjects. Future studies will be necessary to determine if this relationship also applies to T2DM and how gastrointestinal and other factors alter the rate of GSR suppression during an OGTT.

Disclosure

M.C.Laurenti: None. A.Manduca: None. C.Cobelli: None. C.Dalla man: Research Support; Becton, Dickinson and Company, Sanofi-Aventis Deutschland GmbH. A.Vella: Advisory Panel; Crinetics Pharmaceuticals, Inc., Rezolute, Inc., vTv Therapeutics, Zealand Pharma A/S, Other Relationship; Novo Nordisk.

Funding

National Institutes of Health (DK078646, DK116231)
1385-P: Impact of Preanalytical and Analytical Factors on Glucagon and GLP-1 Levels in Humans
Authors: RASMUSSEN CHRISTINE , KJELDSEN SASHA , JENSEN NICOLE J. , HEINZ NIKLAS , HARTMANN BOLETTE , HOLST JENS J. , ALBRECHTSEN NICOLAI J. WEWER
Published: May 2022

Glucagon and glucagon-like peptide-1 (GLP-1) have important roles in the development and treatment of diabetes. Their plasma concentrations are currently being measured in more than 500 clinical studies according to clinicaltrial.gov. As glucagon and total GLP-1 (reflecting the secretion and action of GLP-1 and is analytically independent of DPP-4 activity) circulate in the low picomolar range it has been troublesome to accurately measure their plasma concentrations. Equally important are so-called preanalytical considerations including the use of enzyme inhibitors. The latter may also depend on the analytical approach, for example single-antibody assay versus sandwich ELISA. To provide guidelines for measurement of glucagon and GLP-1 in clinical trials we obtained blood samples using four different blood-containers (with either neprilysin inhibitor: sacubitrilat 25 µg/mL; trasylol 500KIU/mL; P800 tubes; or vehicle) from healthy individuals (male/female; 7/4; mean ± SD; age: 32 ± 12 years, BMI; 26 ± 4 kg/m2) after an overnight fast and following an intravenous amino acid (AA) infusion (14 g/L; 331 mg/min/kg body weight. Plasma concentrations of glucagon increased significantly during the AA-infusion independent of blood containers and assays. The sandwich ELISA showed significant lower levels of glucagon with addition of neprilysin inhibitor whereas both trasylol and P800 tubes resulted in increased levels during the AA-infusion. Glucagon levels measured by the two radioimmunoassays (RIA) did not depend on the containers. Levels of total GLP-1 did not depend on the containers using a sandwich ELISA (Mercodia) but increased by a factor of a 2.5 when plasma from P800 containers was assayed using a C-terminal specific RIA.

In summary, levels of glucagon and GLP-1 depend on both preanalytical and analytical factors that should be standardized for future clinical studies.

Disclosure

S. Kjeldsen: None. N.J. Jensen: None. N. Heinz: None. B. Hartmann: Board Member; Bainan Biotech. J.J. Holst: Advisory Panel; Novo Nordisk. Board Member; Antag Therapeutics, Bainan Biotech. N.J. Wewer Albrechtsen: Research Support; Mercodia AB, Novo Nordisk, Regeneron Pharmaceuticals Inc. Speaker's Bureau; Merck & Co., Inc., Mercodia AB.

Funding

Nicolai J. Wewer Albrechtsen were financed by NNF Excellence Emerging Investigator Grant – Endocrinology and Metabolism (Application No. NNF19OC0055001) , EFSD Future Leader Award (NNF21SA0072746) and DFF Sapere Aude.
1390-P: FOXO1 Promotes Sexually Dimorphic Effects on Energy Balance in the Preoptic Anterior Hypothalamus
Authors: LUO PEI , IRIZARRY VALERIA C. TORRES , YE HUI , IBRAHIMI LUCAS , SCHAUL SARAH , PATEL NIRALI , CARRILLO-SÁENZ LESLIE , ANTONY NIMISHA , DIXIT DEVIN , KOTA MAYA , UNTERMAN TERRY , XU PINGWEN
Published: May 2022

The preoptic anterior hypothalamus (POAH) , known as a thermointegrative center, modulates metabolic adaptation in response to temperature change, thus making it relevant for body weight homeostasis. Forkhead transcription factor O1 (FoxO1) plays an important role in mediating effects of leptin and insulin on metabolic homeostasis in the hypothalamus. However, the homeostatic role of FoxO1 in POAH has not been investigated. We found that FoxO1 specific deletion in the POAH neurons (FoxO1POAH-KO) prevented high fat diet-induced obesity (DIO) in female but not male mice. This female-specific protection was associated with increased lean mass, decreased fat mass, and upregulated thermogenesis in adipose tissues. The female FoxO1POAH-KO mice also showed improved insulin sensitivity and increased energy expenditure when exposed at room temperature (22 °C) or cold (6 °C) . Moreover, the FoxO1POAH-KO-induced protection against DIO in females was blunted by ovariectomy (OVX) , while 17β-estradiol supplementation after OVX failed to restore this protection, suggesting an estrogen-independent mechanism. Finally, mice with FoxO1 constitutively activated in the POAH neurons (FoxO1POAH-CA) significantly increased DIO in both sexes, further supporting an essential metabolic role of FoxO1POAH. Together, these findings indicate that FoxO1POAH is a crucial transcription factor that directs and coordinates control of energy balance, thermogenesis, and glucose homeostasis in female mice.

Key words: FoxO1, POAH, sex difference.

Disclosure

P. Luo: None. H. Ye: None. T. Unterman: None. P. Xu: None.

Funding

P. Xu is supported by National Institutes of Health (R00DK1070 and R01DK123098) , Department of Defense (Innovative Grant W81XWH-19-PRMRP-DA) , and The University of Chicago Diabetes Research and Training Center (The Pilot and Feasibility Award DK020595) .
1405-P: Bacteroides thetaiotaomicron Fortifies the Gut Mucosa and Improves Host Metabolism in Dietary Obese Mice
Authors: LEE JENNIFER , WELLENSTEIN KERRY , KAHN BARBARA B.
Published: May 2022

Disruption of the intestinal mucosa contributes to insulin resistance in obesity and Type 2 diabetes. Probiotic supplementation, which restores gut microbiota composition and fortifies intestinal barrier function, has therapeutic potential to treat obesity and metabolic disease. Treatment of mice on a chow diet or high fat diet (HFD) with Palmitic Acid Hydroxystearic Acids (PAHSAs) improves insulin sensitivity. These effects are mediated, at least in part, by changes in the gut microbiome. We used this paradigm to identify new microbes that may have beneficial metabolic effects. Metagenome sequencing of the cecum from mice treated with PAHSAs identified Bacteroides thetaiotaomicron (Bt) to be most strongly associated with insulin sensitivity. Here, we aimed to determine whether Bt supplementation in mice on a HFD diet has beneficial effects on energy balance and/or glucose homeostasis. Daily oral Bt supplementation in female mice fed a HFD resulted in less weight gain, lower adiposity, and improved glucose tolerance compared to vehicle-treated mice. Additionally, Bt supplementation preserved colonic mucus thickness, an indicator of gut health; increased the number of Goblet cells which are the cells that secrete mucin2, the predominant mucus protein that forms the intestinal mucosal barrier; and increased protein levels of mucin2 in the distal gut compared to vehicle-treated controls. These improvements in distal gut barrier architecture were associated with a reduction in circulating levels of pro-inflammatory lipopolysaccharide (LPS) and interleukin-6 (IL-6) . These beneficial effects of Bt were observed in female and not in male HFD-fed mice. These studies indicate that Bt has sex-specific effects to improve host metabolism in the setting of dietary obesity. Bt represents a next-generation probiotic and novel gut-based therapeutic strategy which could improve metabolic health in women.

Disclosure

J.Lee: None. K.Wellenstein: None. B.B.Kahn: Advisory Panel; Janssen Pharmaceuticals, Inc., Consultant; Mellicell, Other Relationship; Alnylam Pharmaceuticals, Inc.

Funding

NIH KDK114162; JPB Foundation
1417-P: Gut Microbiome Translocation to Visceral Adipose Tissue Promotes Recruitment of Adipose-Specific Neutrophils in Human Obesity
Authors: SHANTARAM DHARTI , BLASZCZAK ALECIA M. , HOYD REBECCA , SMITH ALAN , ANTWI LINDA , LIU JOEY Z. , WRIGHT VALERIE P. , BRADLEY DAVID , LIU YUNZHOU , WILLIAMS NYELIA F. , SPAKOWICZ DAN , HSUEH WILLA
Published: May 2022

Neutrophils protect against invading bacteria, and their unique role in adipose tissue inflammation and insulin resistance (IR) in obesity is increasingly recognized. We found that visceral adipose tissue (VAT) of obese (n=83) compared to lean (n=13) individuals had higher neutrophil abundance, correlating with IR. The obese subjects had elevated circulating plasma zonulin, a marker of intestinal permeability, and lipopolysaccharide-binding protein (LPSBP) , a marker of bacterial presence, consistent with evidence suggesting that changes in the microbiome and gut permeability in obesity permits the entry of gut bacteria and bacterial products into the circulation. Therefore, we tested for bacterial presence in VAT under sterile conditions. 16s rRNA analyses revealed that the family Streptococcaceae and an unnamed genus in the family Ruminococaceae were enriched in obese VAT contrary to order Bacilliales and genus Marvinobryantia in lean VAT, majorly originating from the GI tract. To confirm whether higher bacterial load is causal for VAT neutrophil abundance in obesity, we gavaged high-fat diet fed mice with fecal material from an obese human, which increased neutrophil, but not macrophage recruitment to VAT. As VAT is highly vascularized, we used RNA-seq to compare the gene expression signature of VAT neutrophils to peripheral blood neutrophils to eliminate the possibility that VAT neutrophils were circulating cells. Evidently, VAT neutrophils had a differential activation state compared to the circulating neutrophils, with a distinct gene expression pattern representing upregulated inflammatory, chemotactic, extracellular matrix, and ROS generation pathways. Overall, we find that that increased gut bacterial translocation enhances the recruitment of VAT specific neutrophils in obesity offering novel targets for managing VAT inflammation and comorbidities of obesity.

Disclosure

D.Shantaram: None. N.F.Williams: None. D.Spakowicz: None. W.Hsueh: None. A.M.Blaszczak: None. R.Hoyd: None. A.Smith: None. L.Antwi: None. J.Z.Liu: None. V.P.Wright: None. D.Bradley: None. Y.Liu: None.
1423-P: Endoscopic Revision for Post-Bariatric Weight Regain Results in Less Weight Loss in People with Type 2 Diabetes
Authors: TEJANI SANAA , DUNN SAMUEL H. , TAVAKKOLI ANNA , MESSIAH SARAH E. , ALMANDOZ JAIME
Published: May 2022

Background: Post-bariatric surgery weight regain is common and leads to recidivism of cardiometabolic diseases, including Type 2 Diabetes Mellitus (T2DM) . Endoscopic therapies are emerging tools to treat weight regain. This study aims to assess the effectiveness of endoscopic revision for post-bariatric surgery weight regain in people with T2DM.

Methods: A retrospective review was conducted on patients who underwent endoscopic revision for post-bariatric surgery weight regain from November 20to July 2021 at an academic center. The primary outcome was weight change at 1, 3, and 6 months post-endoscopic revision.

Results: The cohort (n=33) was mean age 55 years; 82% female; 45% non-Hispanic Black, 15% Hispanic; mean BMI 39.3; 63% with gastric bypass, 33% with sleeve gastrectomy, 3% with duodenal switch; 24% with T2DM at the time of revision. The mean time from bariatric surgery to revision was 12 years with mean 33% weight regain from post-surgery nadir. Patients with T2DM experienced similar mean percent weight loss as those without T2DM at 1 (5.34% vs. 4.93%) and 3 months (4.98% vs. 6.47%) post-revision but less weight loss at 6 months (-0.28% vs. 4.07%) (Figure 1) .

Conclusion: Endoscopic revision is an effective therapy for post-bariatric surgery weight regain. However, people with T2DM may experience less long term weight loss compared to those without T2DM, warranting further investigation.

Disclosure

S.Tejani: None. S.H.Dunn: None. A.Tavakkoli: None. S.E.Messiah: None. J.Almandoz: None.
1434-P: ANGPTL8 Variant R59W Is Associated with Increased Circulatory TNF-alpha and IL7 LEVELS and Inflammation
Authors: MOHAMMAD ANWAR , ABU-FARHA MOHAMED , MADHU DHANYA , CHERIAN PREETHI T. , KHAIRI IRINA AL , HEBBAR PRASHANTHA , KAVALAKATT SINA A. , AREFANIAN HOSSEIN , ALAM-ELDIN NADA , THANGAVEL ALPHONSE T. , ALMULLA FAHD , ABUBAKER JEHAD
Published: May 2022

Background: ANGPTL8 has been recently identified as lipid metabolism and inflammation regulator. It interacts with ANGPTL3 and, more recently, ANGPTL4 to regulate LPL activity. It was also shown to modulate NF-κB activity through its interaction with ikkα/β. A genetic variant rs2278426 (R59W) in ANGPTL8 has been associated with reduced LDL and HDL in Hispanics and increased FBG in Arabs. The objective of the study was to investigate the impact of the R59W variant on LPL and the inflammatory activity of ANGPTL8.

Methods: ANGPTL8 was genotyped in a discovery cohort of 738 Kuwaiti individuals. ANGPTL8 level, as well as inflammatory markers and LPL, were measured in plasma. Additionally, metabolite analysis, overexpression, luciferase binding assay, confocal microscopy, and cellular fractionation analyses were conducted in the HepG2 cell line to assess differences between the ANGPTL8 and its variant.

Results: The ANGPTL8 rs2278426 variant was associated with an increase in circulatory levels of TNF-α and IL7 in our cohort. Carrier versus reference genotype of the studied variant showed differences in the following metabolites: Acylcarnitines, Phosphatidylcholine, and Cholesteryl Ester. These metabolites are implicated in modulating NF-κB and TNF-α levels. Our in vitro analyses showed that the activity levels of NF-κB p65 was higher in the R59W than the wild type, and it was further elevated with TNFα stimulations. The data was validated by the luciferase activity, confocal microscopy, and cellular fractionation.

Conclusion: This study showed that ANGPTL8 variant R59W is associated with increased circulatory TNF-α and IL7 levels but not the LPL plasma levels. We have also demonstrated the pro-inflammatory role of the R59W variant in regulating the NF-κB pathway as reflected by genotyping, in vitro, metabolomics, and structural analyses. Further studies are needed to expand our understanding of other signaling pathways and possible crosstalk in the context of inflammation.

Disclosure

A.Mohammad: None. A.T.Thangavel: None. F.Almulla: None. J.Abubaker: None. M.Abu-farha: None. D.Madhu: None. P.T.Cherian: None. I.Al khairi: None. P.Hebbar: None. S.A.Kavalakatt: None. H.Arefanian: None. N.Alam-eldin: None.

Funding

Kuwait Foundation for the Advancement of Sciences
144-LB: Pediatric Type 2 Diabetes Prevalence from Real-World Data in the United States
Authors: CHAO LILY , VIDMAR ALAINA , ESPINOZA JUAN
Published: May 2022

Background: The SEARCH for Diabetes in Youth Study has provided critical insights into the epidemiology of pediatric T2D. The 2021 report estimated an annual percent change (APC) in prevalence of 4.8% between 2009 to 2017. Multi-center longitudinal cohort studies generate robust data, but are time, resource, and effort intensive. Large real-world data repositories may offer cost-effective opportunities to validate and expand on these findings.

Hypothesis: Youth-onset T2D prevalence is rising at a rate in excess of 5% per year in the US in the previous five years.

Methods: We queried the Trinetx Research Network which includes EHR data from 58 institutions with 86,487,854 total patients. We included patients <22 years of age, who had at least one incidence of an E11 ICD-10 code for T2D associated with an ambulatory, emergency room, or inpatient encounter for each calendar year from 2016 to 2021. Patients with other diabetes ICD-10 codes (E08, E09, E10, or E13) were excluded. Summary statistics of the query cohort describing sex, race and ethnicity, labs, medications, and anthopometrics were extracted from the platform. Chi-squared test for trend was used to determine statistical significance in demographic change.

Results: The mean APC in prevalence per center rose by 9.1% between 2016 to 2019 (2016: 127 cases/center, 5334 total cases; 2019: 166.2 cases/center, 7919 total cases) , but subsequently declined by 6.6% between 2019 to 2020. There was a consistent increase in the median BMI percentile over time from 2016 to 2021. The percentage of black youth diagnosed with T2D rose (from 30 to 33%, P<0.0001) whereas that of Hispanic youth was unchanged (25%, P=0.73) . The proportion of male also increased over time (P<0.0001) .

Conclusion: Real-world data from Trinetx demonstrates a rising incidence of youth-onset T2D that parallels an increase in median BMI percentile. The decrease in case rate in 2020 may be the result of decreased encounters during the Covid-19 pandemic. The rising prevalence of T2D in black youth and male gender warrants further analysis, to understand the driver behind this trend.

Disclosure

L. Chao: None. A. Vidmar: None. J. Espinoza: Consultant; AI Health.

Funding

National Center for Advancing Translational Science (UL1TR001855 and UL1TR000130)
1444-P: Knockdown of Etv5 Impairs Viability of Pancreatic Beta Cells by Enhancing ROS Production
Authors: YI XUANZI , BAGHNAVI FAHIMEH , DIEKMANN CEDRIC , PÄTH GÜNTER , SEUFERT JOCHEN
Published: May 2022

Introduction: The transcription factor ETV5 regulates various biological processes such as cell proliferation and apoptosis, and activates a whole string of obesity- and diabetes-related genes. ETV5 is involved in tissue protection since mice with pancreatic loss of Etv5 show increased severity of pancreatitis and delayed tissue recovery. Here, we investigate the role of Etv5 knockdown (KD) on survival of injured endocrine pancreatic beta cells.

Methods: We generated INS-1E beta cells with doxycycline (dox) -inducible lentiviral Etv5 sh-RNA knock-down (KD) . Cellular stress was induced by streptozotocin (STZ) and palmitate (PA) . We assessed viability (MTS) , apoptosis (caspases-3/7 activation) , necrosis (assay) , ER stress (BiP and CHOP gene expression) , oxidative stress (ROS production) and gene expression of SOD1, SOD2 and Catalase.

Results: Etv5 KD significantly increases STZ/PA-induced loss of viability without changes in apoptosis but with increased necrosis. Etv5 KD did not change STZ/PA-induced gene expression of ER stress markers BiP and CHOP but significantly increased H2O2 production. This was associated with significantly elevated gene expression of mitochondrial SOD2, which converts superoxide into H2O2. Cytoplasmatic SOD1 and H2O2-decomposing catalase, PRDX1 and GPx1 remained unaffected.

Conclusion: Increased circulating free fatty acids in obesity promote oxidative stress. Loss of ETV5 increases ROS production in mitochondria during cellular stress by PA and STZ, as indicated by upregulated SOD2 gene expression. This suggests a role for ETV5 in controlling mitochondrial oxidative stress in pancreatic β-cells.

Disclosure

X. Yi: None. G. Päth: None. J. Seufert: Advisory Panel; Abbott, Sanofi-Aventis Deutschland GmbH. Research Support; Boehringer Ingelheim International GmbH. Speaker's Bureau; Abbott Diabetes, AstraZeneca, Bayer AG, Boehringer Ingelheim International GmbH, Lilly, Novo Nordisk, Sanofi-Aventis Deutschland GmbH.

Funding

XY is supported by China Scolarship Council (CSC) China,GP received Projektförderung from the German Diabetes Society (DDG)
1448-P: Partial Pancreatectomy Increased α-Cell Volume Density, but Not Compensated ß-Cell Volume in Japanese Subjects
Authors: TAKEUCHI YUKI , MIZUKAMI HIROKI , OSONOI SHO , KUDOH KAZUHIRO , SASAKI TAKANORI , YAGIHASHI SOROKU
Published: May 2022

In type 2 diabetes, deficit of insulin secretion is often ascribed to the reduction of β-cell volume, implying that regeneration of β-cells could be one of its radical treatments. Hitherto, several models are known to elicit β-cell compensation in rodents. Among them, application of partial pancreatectomy can induce replenishment of β-cell volume in young mice, while it is unclear whether this occurs in humans. In the present study, islet pathology was evaluated using the pancreatic specimens from the patients with pancreatic cancer who underwent repeated (i.e., partial followed by total) pancreatectomy because of its local recurrence after the initial surgery. Ten subjects (5 males and 5 females) at Hirosaki University Hospital or Chiba University Hospital were evaluated. Morphometric analysis was performed on formalin fixed pancreatic sections, in which the quadruple immunostaining for each hormone and the fluorescent immunostaining for ALDH1A3 was conducted. The mean age at the first and second operations was 66.2±and 71.6±5.8 years old, respectively. The mean duration until recurrence was 3.2±2.0 years. There were no significant differences in HbA1c and BMI between first and second preoperation. In morphometrical analysis, α-cell volume in the second surgery was significantly increased compared to the first surgery (p<0.04) , while β-cell volume was comparable. There was a positive correlation between the frequency of conduit glucagon positive cells and islet α-cell volume (r=0.66, p<0.05) . The frequency of ALDH1A3 positive cells were positively correlated with the α-cell/ β-cell ratio in the second surgical specimen (r=0.81, p<0.01) . Taken together, unlike rodents, our results indicate that partial pancreatectomy did not induce replenishment of β-cell volume, but increased α-cell volume in human subjects. As its mechanism, the neogenesis of ductal endocrine cells and/or the transdifferentiation of β-cells to α-cells in the islets may be involved.

Disclosure

Y.Takeuchi: None. H.Mizukami: None. S.Osonoi: None. K.Kudoh: None. T.Sasaki: None. S.Yagihashi: None.
1450-P: Identifying Hidden Sources of Tissue Plasticity in Pancreatic Islets with a Novel Multicolor Flow Cytometry Approach
Authors: KRYVALAP YURY , MENG SHAWN Z. , CZYZYK JAN
Published: May 2022

Identifying pancreatic islet cells that acquire a beta-cell like phenotype is important for developing new diabetes therapies. Our studies have revealed a novel serpinB13 protease inhibitor antibody, that augments cathepsin L protease activity, resulting in Notch1 receptor cleavage and mobilization of endocrine progenitor cells to become beta cells in the embryonic pancreas. It is unclear, however, whether a similar induction of additional beta cells occurs with exposure to serpinB13 monoclonal antibody (mAb) in later life.

To explore the exact source (s) of new beta cells in response to serpinB13 mAb after birth we developed a novel, multicolor flow cytometry analysis approach to study pancreatic islets. Five-week old normal Balb/c mice were injected with serpinB13 mAb (or control IgG) , and sacrificed at 8 weeks for islet isolation. Islet cell suspensions were stained with a viability dye, and a panel of antibodies to CD31, CD45, cytokeratin 19, EpCAM, insulin (Ins) , glucagon (Glu) , somatostatin (SS) , c-peptide, and four transcription factors, Maf-A, Nkx6.1, NeuroD1, and Pdx-1.

Exposure to the serpinB13 mAb caused a significant increase in the number of hormonal-negative (e.g., Ins-Glu-SS-c-peptide-) cells (p=0.0011) , and hormonal negative, but c-peptide positive, cells (p=0.002) . At least one of four transcription factors examined were positive in 5-10% and 10-30% of these populations, respectively. At the same time, there was a trend in the drop of Glu+ and SS+ cells. The average number of pancreatic islets, and the total islet cellularity per mouse, was augmented in animals receiving serpinB13 mAb.

Although additional studies are necessary to characterize the hormonal negative cells and assess whether Glu+ and SS+ cells transdifferentiate to beta cells, these findings demonstrate that enhanced tissue plasticity and beta-cell output can be achieved in the pancreas following treatment with antibodies to protease inhibitors.

Disclosure

Y.Kryvalap: None. S.Z.Meng: None. J.Czyzyk: None.
1459-P: Human Islet Cell Transcriptome Analysis Using Single Nucleus RNA-seq Reveals New Beta-Cell Markers and Define Distinct Beta-Cell Subpopulations with Different Transcriptional Activity
Authors: KANG RANDY , LI YANSUI , ROSSELOT CAROLINA , SCOTT DONALD , GARCIA-OCANA ADOLFO , LU GEMING
Published: May 2022

Single-cell RNA sequencing (scRNA-seq) of human islets requires cell dissociation and does not provide information on the transcriptional status of islet cells. On the other hand, single-nucleus RNA sequencing (snRNA-seq) does not require cell dissociation and provides abundant information on intronic sequences that can be used to identify actively transcribed genes. Based on this, we seek to compare scRNA-seq and snRNA-seq approaches in human islets to determine: 1) whether similar cell clusters could be detected; 2) whether new gene markers could be discovered in human islet endocrine cells using intronic reads; 3) whether human beta cell subpopulations could be identified based on INS expression dynamics; and 4) whether snRNA-seq could be used in human islet grafts transplanted in immunosuppressed mice. We performed scRNA-seq and snRNA-seq on three pairs of human islets obtained from three healthy adult human donors using exon only or exon plus intron reads, respectively. Analysis of integrated data revealed similar human islet cell clusters. In the snRNA-seq data, however, top differentially expressed genes were identified as new markers of human endocrine cells such as ZNF385D (β-cells) , PTPRT (α-cells) , LRFN5 (δ-cells) and CACNA2D3 (PP cells) . These markers also accurately define endocrine cell populations in human islet grafts. Additionally, we distinguished several beta cell sub-clusters- INS rich cluster, HNRNPA2B1 (vesicle) rich cluster and active INS transcribing cluster.

In conclusion, snRNA-seq analysis of human islet cells is a previously unrecognized tool for the identification of human islet cell types in samples where nuclear RNA processing is required. By comparing INS expression between scRNA-seq and snRNA-seq data sets, we can detect different beta cell sub-populations with distinct gene expression patterns representing different biological dynamic states.

Disclosure

R.Kang: None. Y.Li: None. C.Rosselot: None. D.Scott: None. A.Garcia-ocana: Consultant; Sun Pharmaceutical Industries Ltd. G.Lu: None.

Funding

NIH DK105015
1460-P: Tauroursodeoxycholic Acid (tudca) Improves the Morphofunction of Pancreatic Beta Cells in Postweaning Protein-Restricted Mice
Authors: SANTOS LOHANNA M. BARRETO DOS , ARAUJO THIAGO , ALVES BRUNA L. , FERREIRA SANDRA MARA , ZIMATH PRISCILA L. , VETTORAZZI JEAN F. , CARNEIRO EVERARDO M.
Published: May 2022

Protein restriction in the postweaning diet in mice impairs glucose metabolism by reducing insulin secretion from pancreatic β cells. TUDCA in obese rodents and humans normalized insulin secretion, increased insulin clearance, furthermore recovered mass and the number of β cells per islet in animal models of diabetes. This study aimed to evaluate the effects of TUDCA on the morphology and function of pancreatic β cells in mice subjected to postweaning protein restriction. Male C57Bl/6J mice at 30 days of age received a control diet (14% protein - group C) or a protein-restricted diet (6% protein - group R) for 16 weeks. During the last 2 weeks, 50% of the animals in each group received intraperitoneal treatment with daily doses of 300 mg/kg of body weight of TUDCA (CT and RT) , and the remainder received PBS (C and R) . The R mice that showed decreased total plasma protein and serum albumin concentration had lower weight gain and feed efficiency, but increased white adipose tissue weight. There were no differences in fasting blood glucose between the groups. The R mice that have increased insulin sensitivity after treatment with TUDCA had normalized sensitivity. Isolated islets from R mice have lower insulin secretion, which has been associated with decreased morphometric parameters compared to C. In RT mice secretion was restored in response to 2.8 mM and 11.mM glucose and the pancreas of the RT showed a greater area of β cells in relation to the area of the islets and a greater number of islets compared to R. Furthermore, RT mice showed an increase in the expression of genes of the SNARE complex, genes of function, transcription and differentiation of pancreatic β cells. We conclude that TUDCA improves glycemic homeostasis, increases insulin secretion and improves the functioning of pancreatic β cells in mice subjected to protein restriction, indicating that TUDCA can be used as a therapeutic strategy to normalize insulin secretion in protein malnutrition.

Disclosure

L.M.Barreto dos santos: None. T.Araujo: None. B.L.Alves: None. S.Ferreira: None. P.L.Zimath: None. J.F.Vettorazzi: None. E.M.Carneiro: None.

Funding

Conselho Nacional de Desenvolvimento Científico e Tecnológico (134270/2019-3)
1461-P: Quantifying Proinsulin Misfolding in the Endoplasmic Reticulum
Authors: ARUNAGIRI ANOOP , ALAM MD MAROOF , HAATAJA LEENA , SAMY PRAVEEN J. , SOLEIMANPOUR SCOTT , SATIN LESLIE S. , ITKIN-ANSARI PAMELA , LIU MING , ARVAN PETER
Published: May 2022

We recently reported that normal pancreatic islets or beta cell lines contain a subset of proinsulin molecules that have not achieved the three intramolecular disulfide bonds required for native insulin structure. Indeed, we found that non-native proinsulin monomers can form intermolecular disulfide-linked complexes, which are detectable by immunoblotting after nonreducing SDS-PAGE. However, we have recognized difficulties in the quantitative interpretation of these analyses, as we frequently identified conditions in which little or no proinsulin monomer was detected, and non-native disulfide-linked complexes amounted to greater cumulative proinsulin band intensity than the entire population of proinsulin molecules detected by immunoblotting upon reducing SDS-PAGE. Here we have developed several critical modifications of our methodology to rectify these discrepancies to undertake a more accurate determination of the pool of misfolded proinsulin molecules in pancreatic beta cells under conditions of health and disease. Using these modifications, we find that the size of the proinsulin monomer pool has previously been underestimated and the fraction of disulfide-linked complexes overestimated — and this can be readily explained by proinsulin antibody affinity in the immunoblotting protocol. Additionally by immunoblotting, we now resolve native and non-native proinsulin monomers, highlighting the inefficiency of non-native proinsulin transport through the secretory pathway.

Disclosure

A.Arunagiri: None. M.Alam: None. L.Haataja: None. P.J.Samy: None. S.Soleimanpour: None. L.S.Satin: None. P.Itkin-ansari: None. M.Liu: None. P.Arvan: n/a.

Funding

National Institutes of Health (R01-DK48280) , National Institutes of Health (U01-DK127747) , and National Institutes of Health (R24 DK110973)
1466-P: Pancreatic Delta Cells Switched Islet's Ca2+ Oscillation Modes by Inhibiting Endogenous Glucagon from Alpha Cells
Authors: REN HUIXIA , GROUP CHAO TANG , GROUP LIANGYI CHEN
Published: May 2022

Glucose-stimulated pancreatic islets show different patterns of Ca2+ signaling with fast, slow and mixed fast/slow oscillations. However, it is not clear whether they are characteristic features of individual islets. We report islet spontaneous slowdown from fast to slow/mixed Ca2+ oscillation under high glucose stimulation. The fast oscillation stage typically lasts for several hours and intrinsic to the islet. The modes switch is reversible. Low glucose resting for 30 minutes fully restored the fast oscillation. Microfluidic perfusion delays and static incubation brings forward the mode switch, which suggests the critical role of the paracrine environment. Correspondingly, exogenous glucagon delays and somatostatin brings forward the mode switch. Pharmacologically blocking Sstr2 on α cells triggers endogenous glucagon secretion and switches islets back to fast oscillations. Optoinhibition and relief of δ cells cause islets to repeatably show fast and slow oscillations. Our study highlights the importance of δ-α cell interactions to generate tunable islet oscillation patterns. The design principle of pancreatic islet was revealed, as the minority of δ cells command the majority of α and β cells.

Funding

No
149-LB: Pediatric Exogenous Insulin Antibody Syndrome—Successful Treatment with Mycophenolate Mofetil
Authors: SABA LESLIE , ORANDI AMIR B. , PITTOCK SIOBHAN , CREO ANA
Published: May 2022

Case Description: A lean, 17-year-old Caucasian male with type 1 diabetes diagnosed at age 9 experienced an abrupt increase in his insulin requirements. He was managed on mixed daily injections. Daily insulin requirements increased from 1.7 units/kg/day to 3.3 units/kg/day over 6 months. His total insulin level was 7 mcIU/mL with a free insulin level of 4.8 mcIU/mL, indicating exogenous insulin antibodies. Switching from insulin aspart to glulisine, due to presumed antibodies against insulin aspart was unsuccessful, and his insulin requirements continued to increase to 4.4 units/kg/day. Immunosuppressive therapy was commenced with mycophenolate mofetil, decreasing his insulin requirements to 1.1 units/kg/day after 7 months of treatment. Repeat testing revealed an improvement in the discrepancy between total and free insulin levels, with a total insulin level of 5.2 mcIU/mL and free insulin level of 4.6 mcIU/mL.

Discussion: Exogenous insulin antibody syndrome should be considered in patients with poorly controlled diabetes and requiring greater than 2 units/kg/day of insulin without obesity. Classically, antibodies against exogenous insulin were found in patients receiving insulin derived from animal sources; however, recombinant human insulin can very rarely trigger an antibody response in select patients. Immunosuppression using mycophenolate mofetil greatly reduced insulin requirements and this case highlights its novel use in children with diabetes who develop insulin antibody syndrome.

Disclosure

L. Saba: None. A. B. Orandi: None. S. Pittock: None. A. Creo: None.