bluuebunny/crossref_metadata_2025 huggingface dataset:
1906-P: Glucose-Dependent Insulinotropic Polypeptide (GIP) Reduces Bone Resorption in Patients with Type 2 Diabetes
Authors: CHRISTENSEN MIKKEL B. , LUND ASGER , JØRGENSEN NIKLAS R. , HOLST JENS J. , VILSBØLL TINA , KNOP FILIP K.
Published: June 2020

In healthy individuals, GIP enhances insulin secretion and reduces bone resorption As GIP has impaired insulinotropic effects in patients with type 2 diabetes (T2D), we investigated the effect of glucose and GIP on bone resorption in these patients.

In a randomized, double-blinded, cross-over study, 12 male patients with T2D underwent six study days with 90-minute GIP or matching placebo (saline) infusions on paired days with plasma glucose (PG) clamped at three levels: Fasting hyperglycemia (mean PG ∼8 mmol/l), aggravated hyperglycemia (mean PG ∼12 mmol/l), or insulin-induced hypoglycemia (mean PG 3-8 mmol/l). Primary outcome measure was CTX (a marker of bone resorption expressed as %-change from baseline ± SD).

On days with fasting hyperglycemia, CTX was suppressed by 36±15% during GIP compared to 0±9% during saline infusion (p<0.0001). On days with aggravated hyperglycemia, CTX was suppressed by 47±23% during GIP compared to 10±9% during saline infusion (p=0.0005). On days with insulin-induced hypoglycemia, CTX was suppressed by 40±15% during GIP compared to 12±11% during saline infusion (p<0.0001).

In conclusion, Short-term GIP infusions during hyperglycemia reduce bone resorption by more than a third suggesting that GIP has preserved effects on bone in patients with T2D.

Disclosure

M.B. Christensen: None. A. Lund: Speaker’s Bureau; Self; AstraZeneca, Novo Nordisk A/S, Sanofi. N.R. Jørgensen: None. J.J. Holst: Advisory Panel; Self; AstraZeneca, Merck Sharp & Dohme Corp., Novo Nordisk A/S, Zealand Pharma A/S. Other Relationship; Spouse/Partner; Antag Therapeutics. T. Vilsbøll: Advisory Panel; Self; AstraZeneca, Mundipharma International, Novo Nordisk A/S, Sun Pharmaceutical Industries Ltd. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Lilly Diabetes, Medscape, Merck Sharp & Dohme Corp., Sanofi. F.K. Knop: Advisory Panel; Self; AstraZeneca, Merck Sharp & Dohme Corp., Mundipharma International, Novo Nordisk A/S, Sanofi. Consultant; Self; Carmot Therapeutics, Inc., Eli Lilly and Company, Novo Nordisk A/S. Research Support; Self; AstraZeneca, Gubra, Novo Nordisk A/S, Sanofi, Zealand Pharma A/S. Speaker’s Bureau; Self; AstraZeneca, Lupin Pharmaceuticals, Inc., Merck Sharp & Dohme Corp., Norgine B.V., Novo Nordisk A/S.
1945-P: Pioglitazone Promotes the Function of White and Brown Adipose Tissue in Diet-Induced Obese Mice
Authors: XU FEN , WANG WEI , ZHENG XIAOBIN , GUO WANRONG , SHEN YUNFENG
Published: June 2020

PPARγ agonist pioglitazone is an insulin sensitizer in the treatment of type 2 diabetes. Here, we investigate the effects of pioglitazone on white and brown fat in diet-induced obese mice. After 16-week diet intervention, C57BL/6 mice were divided randomly into three groups: normal chow diet group (NC group), high-fat diet group (HFD group) and high-fat diet + pioglitazone group (HFDP group). Pioglitazone (30-40mg/kg/day) was administered in drinking water for 6 weeks. After 6-week pioglitazone treatment, there was no significant difference in both body weight and fat mass between the HFDP group and the HFD group. However, fasting blood glucose, glucose tolerance and insulin sensitivity were significantly improved in HFDP group. What’s more, the weight of liver and epididymal adipose tissue (eWAT) were not reduced by pioglitazone treatment. However, the weight of brown adipose tissue (BAT) in the HFDP group was significantly higher than HFD group. Histological staining showed that the hepatic steatosis was ameliorated by pioglitazone. Quantitative real-time PCR revealed that mRNA levels of heat-producing genes (PRDM16, FGF-21, UCP1) and mitochondrial biosynthesis and functional genes (Tfam, cidea, cox7a1, Fabp4) were upregulated in eWAT by pioglitazone. Western blotting results also showed that the protein expression of UCP1 and PGC1α was upregulated by pioglitazone both in eWAT and in BAT when compared with the HFD group. To sum up, our study revealed that pioglitazone could promote the browning of WAT and activate the function of BAT with improved hepatic steatosis in diet-induced obese mice.

Disclosure

F. Xu: None. W. Wang: None. X. Zheng: None. W. Guo: None. Y. Shen: None.
1946-P: Dapagliflozin Suppresses Adipose Fatty Acid Accumulation in Rats Fed on High-Fat Diet but Not on Normal Chow
Authors: SATO DAISUKE , AMARUME JOTA , YANO MIZUNA , UMEHARA YUTA , KUSUNOKI MASATAKA , FENG ZHONGGANG , NAKAMURA TAKAO
Published: June 2020

SGLT2 inhibitor (SGLT2i) exerts not only on enhancement of glucose excretion but also on improvement of serum lipid profile via increase in lipid utilization as an energy source instead of glucose. That is, SGLT2i might affect the lipid storage. In the present study, we evaluated the effects of dapagliflozin (DAPA) on weight and fatty acid contents of adipose tissues.

At 8 weeks of age, male rats were divided into four groups: (a) NC (n=6) fed on a standard laboratory chow (LC); (b) NC+DAPA (n=6) on the LC with 1 mg/kg/day DAPA administration; (c) HFD (n=6) on a high-fat diet (HF); (d) HFD+DAPA (n=6) on the HF with 1 mg/kg/day DAPA administration. After 7 weeks, we weighed epididymal (EPI), mesenteric (MES), retroperitoneal (RET), and inguinal subcutaneous (SUB) adipose tissues, and measured 21 adipose fatty-acid contents (μmol/g BW).

As a result, adipose tissue weight in the examined four sites in HFD rats were significantly higher (P<0.01) than those, respectively, in NC rats. No significant differences were observed in weights of any adipose tissues or in their contents of fatty acids between the NC and NC+DAPA rats. In contrast, MES weight in HFD+DAPA rats was significantly lower (P<0.05) than that in the HFD rats. Contents of 1, 3, 2 and 1 saturated fatty acids (SFAs) in EPI, MES, RET and SUB, respectively, in the HFD+DAPA rats were significantly lower (P<0.05) than those, respectively, in the HFD rats. Contents of 3 and 2 monounsaturated fatty acids in MES and SUB, respectively, and contents of 2 and 1 polyunsaturated fatty acids in MES and RET, respectively, in the HFD+DAPA rats were significantly lower (P<0.05) than those, respectively, in the HFD rats.

These results suggest that DAPA could reduce some SFAs in any adipose tissues in HF-fed rats, but not in LC-fed rats. The reduction might sometimes arise without concomitant decrease in tissue weight. In addition, DAPA might suppress accumulation of a wide range of fatty acids in MES of HF-fed rats in particular, probably due to their consumption.

Disclosure

D. Sato: None. J. Amarume: None. M. Yano: None. Y. Umehara: None. M. Kusunoki: Other Relationship; Self; Kaken Pharmaceutical Co., Ltd., Ono Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd. Z. Feng: None. T. Nakamura: Research Support; Self; AstraZeneca K.K., Ono Pharmaceutical Co., Ltd.

Funding

Ono Pharmaceutical Co., Ltd.; AstraZeneca K.K.
1962-P: Role of Transcription Factor TCF21 in Adipocyte Progenitor Cells of Visceral Fat
Authors: MINAMIZUKA TAKUYA , MAEZAWA YOSHIRO , UDAGAWA HARUHIDE , BABA YUSUKE , KOSHIZAKA MASAYA , ENDO YUSUKE , YASUDA KAZUKI , YOKOTE KOUTARO
Published: June 2020

Accumulation of visceral fat leads to insulin resistance and diabetes. Therefore, it is important to elucidate the mechanism by which adipose progenitor cells (APCs) are maintained and how they differentiate. The bHLH transcription factor 21 (Tcf21) is expressed in visceral, but not in subcutaneous fat, and used as a marker for white adipose tissues. Although Tcf21 promotes IL6 expression in APCs in vitro, little is known about its role in vivo. Therefore, we aimed to clarify the role of Tcf21 in visceral fat using mouse models. Tcf21 was deleted after birth using inducible Cre-LoxP system to circumvent the lethality due to cardiac malformation (iTcf21 KO). The mice were fed high fat diet (HFD) from 6 to 24 weeks of age. Weight gain, blood glucose level, epididymal fat weight, and APC count (0.8x104/g fat for iKO and 2.9x104/g fat for wild type (WT)) of epididymal fat were all low in iTcf21KO compared to WT mice. Insulin sensitivity was increased in iTcf21KO mice. Staining of epididymal fat from Tcf21lacZ/+ mice using X-Gal revealed that Tcf21 was localized in the vascular mural cells. Isolation of Sca1+ Lin- APCs using magnetic activated cell sorting revealed that Tcf21 mRNA was expressed in the APCs, along with Pdgfrb and Periostin. In contrast, Tcf21 expression was attenuated in the adipose tissue of WT mice after HFD, and in adipocyte culture that was differentiated from the vascular stromal fraction (SVF). In addition, Tcf21 expression suppressed by HFD was upregulated in adipocytes undergoing de-differentiation by ceiling culture. Adipocyte induction from SVF resulted in no differences between WT and iTcf21KO mice assessed by oil red O staining. However, adipocyte differentiation and expression of Pparg and Cebpb were suppressed in 3T3-L1 cells in which Tcf21 was overexpressed. These findings suggest that Tcf21 is expressed in the APCs of vascular mural cells, controls the number of APCs, and potentially contributes to the maintenance of their progenitor state through suppression of adipocyte differentiation.

Disclosure

T. Minamizuka: None. Y. Maezawa: None. H. Udagawa: None. Y. Baba: None. M. Koshizaka: None. Y. Endo: None. K. Yasuda: None. K. Yokote: Research Support; Self; Astellas Pharma Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Mochida Pharmaceutical Co., Ltd., Nippon Boehringer Ingelheim Co. Ltd., Novartis Pharma K.K., Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Shionogi & Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Teijin Pharma Limited. Speaker’s Bureau; Self; Abbott, Astellas Amgen, AstraZeneca K.K., Bayer Inc., FUJIFILM Pharmaceuticals U.S.A., Inc., Kaken Pharmaceutical Co., Ltd., Sanwa Kagaku Kenkyusho.
1966-P: Induction of Beige Adipocyte Markers in 3T3-L1 Cells by Clk1 and PKCbetaII Inhibitors
Authors: PATEL ACHINTYA , DOBBINS TRADD , KONG SONYA , PATEL NIKETA A. , COOPER DENISE R.
Published: June 2020

Excessive dietary intake of fat results in its storage in white adipose tissue (WAT), whereas energy expenditure by lipid oxidation occurs in brown adipose tissue. A third type of fat is beige, and it is observed in WAT depots. Certain WAT can undergo beiging. Numerous factors have roles in beiging. However, little is known about the signaling pathways inducing pre-adipocytes to become beige adipocytes. Known participants are activators of PPARgamma, PGC1alpha, and PRDM16 that induce UCP1-containing mitochondria upon cold exposure. We found a signaling pathway that regulates alternative pre-mRNA splicing is also involved in beiging when it is inhibited. Clk1 kinase phosphorylates proteins with serine and arginine rich domains. Clk1 inhibition by 50 nM TG003 during differentiation of 3T3-L1 adipocytes (post day 3) results in beige-like adipocytes demonstrating increased PGC1alpha and UCP1 mRNA and proteins. TG003 inhibits Clk1, 2, and 4. We demonstrate using siRNA for Clk1, 2, and 4, that Clk1 inhibition increased UCP1, PGC1alpha and PPARgamma mRNA whereas Clk1 and 4 siRNA did not. TG003-treated adipocytes contained fewer lipid droplets and more mitochondria which resulted in significant proton leak. In addition to TG003, CG53353, a PKCbetaII inhibitor, also increased mitochondria number and decreased lipid droplet size. PKCbetaII is an mRNA splice variant regulated by Clk1. Others previously showed that TG003 treatment of mice on high fat diets (HFD) resulted in weight loss, and PKCbeta(I+II) knockout mice did not gain weight on HFD. Using bioinformatics, we determined that PGC1alpha was a substrate for Clk1 and PKCbetaII. PGC1alpha was shown to be phosphorylated by Clk2 previously. Hence, we believe that inhibition of Clk1 and PKCbetaII phosphorylation of PGC1alpha results in upregulation of UCP1 expression in adipocytes. This newly identified pathway supports a role for Clk1 and PKCbetaII as potential targets for inhibition to treat weight loss in obesity.

Disclosure

A. Patel: None. T. Dobbins: None. S. Kong: None. N.A. Patel: None. D.R. Cooper: None.

Funding

U.S. Department of Veterans Affairs (5|01BX003689)
1992-P: Metabolic Factors Associated with Cognitive Function and Brain Morphology in Type 2 Diabetes
Authors: HENN ROSEMARY E. , REYNOLDS EVAN L. , PATTERSON ADAM , NELSON ROBERT G. , FELDMAN EVA L. , CALLAGHAN BRIAN C.
Published: June 2020

Background: Pima Indians from the Gila River Indian Community in Arizona, USA, have a high prevalence of type 2 diabetes and have been studied for decades to evaluate the risk factors for and trajectory of diabetic complications. Here, we aimed to identify metabolic factors associated with central nervous system sequelae in this population.

Methods: Cognitive performance and ultrastructural brain volumes were determined in Pima Indians via the NIH Toolbox Cognitive Battery (NIHTB-CB) (n=51) and MRI studies (n=40). The NIHTB-CB Composite Score was the primary measure of cognitive function, adjusting for age, sex, race, and education level. MRI derived morphology measures included total cortical volume and white and gray matter volumes. Metabolic factors included duration of diabetes, height, weight, BMI, SBP, DBP, HbA1c, triglycerides, HDL, LDL, and total cholesterol. We performed univariate linear regressions on MRI outcomes and NIHTB-CB Composite Scores as functions of metabolic factors after adjusting for age/sex, and we calculated Pearson’s correlation between each MRI morphology outcome and the Composite Score.

Results: Of the 51 subjects who underwent the NIHTB-CB, 40 completed an MRI. Linear regression models revealed that only BMI associated with both total cortical volume (BMI point estimate (PE): -.0007, CI: -.001, -.0002) and total gray matter volume (BMI PE:-.0009, CI: -.001, -.0003). No metabolic factors associated with the NIHTB-CB Composite Score, and the NIHTB-CB Composite Score did not correlate with any morphology measures.

Conclusion: Increased BMI was associated with both decreased cortical volume and gray matter volume in this Pima cohort, similar to observations found for obesity in previously published cohorts. Though volume measures and NIHTB-CB scores did not correlate in this study, future work will analyze additional MRI based morphology measures to evaluate correlations with individual NIHTB-CB tests and assess changes over time.

Disclosure

R.E. Henn: None. E.L. Reynolds: None. A. Patterson: None. R.G. Nelson: None. E.L. Feldman: Consultant; Self; Novartis Pharmaceuticals Corporation. B.C. Callaghan: None.

Funding

National Institutes of Health (R24DK082841, T32NS0007222); Robert and Katherine Jacobs Environmental Health Initiative; Robert E. Nederlander, Sr. Program for Alzheimer’s Research; National Institute of Diabetes and Digestive and Kidney Diseases; Sinai Medical Staff Foundation
2004-P: Treatment with Testosterone Undecanoate Injections (TU) Results in Marked Weight Loss That Is Maintained Long Term in Men with Hypogonadism and Type 2 Diabetes (T2DM): Update of Real-World Data from a Urological Practice
Authors: HAIDER AHMAD , HAIDER KARIM S. , SAAD FARID
Published: June 2020

Background: The prevalence of hypogonadism is particularly high in men with obesity and T2DM.

Methods: Of 356 men with hypogonadism and T2DM, 288 (80.9%) were obese, 61 (17.1%) overweight and 7 (2.0%) had normal weight. 178 received TU 1000 mg/12 weeks (T-group) following an initial 6-week interval, 178 opted against treatment (CTRL). Changes over time between groups were compared and adjusted for age, weight, WC, fasting glucose, blood pressure, lipids and quality of life to account for baseline differences between groups.

Results: Mean baseline age: 62.6±5.2 years, mean follow-up: 8.2. T-group: weight decreased by 22.1±0.5 kg at 11 years. CTRL: weight increased by 6.8±0.5. Estimated adjusted difference between groups: 28.9 kg (p<0.0001 for all). T-group: WC decreased by 13.3±0.4 cm. CTRL: WC increased by 7.1±0.4 cm. Difference between groups: 20.4 cm (p<0.0001 for all). T-group: BMI decreased by 7.3±0.2 kg/m². CTRL: BMI increased by 2.2±0.2 kg/m². Difference between groups: 9.5 kg/m² (p<0.0001 for all). There was a weight loss of 19.3±0.4% in the T-group and weight gain of 7.4±0.4% in CTRL, difference between groups: 26.7% (p<0.0001 for all). Waist:height ratio decreased by 0.08±0.00 in the T-group and increased by 0.04±0.00 in CTRL, difference between groups: 0.12 (p<0.0001 for all). Visceral adiposity index (VAI) decreased by 3.0±0.2 in the T-group and increased by 3.2±0.2 in CTRL, difference between groups: 6.2 (p<0.0001 for all). Since injections were administered in the office and documented, there was a 100% adherence to testosterone therapy.

Conclusions: Treatment with testosterone undecanoate injections results in profound weight loss that is maintained long-term with continued testosterone therapy.

Disclosure

A. Haider: None. K.S. Haider: None. F. Saad: Consultant; Self; Bayer AG. Stock/Shareholder; Self; AbbVie Inc., Bayer AG.

Funding

Bayer AG
2028-P: Adding Exercise to Liraglutide Treatment in Weight Loss Maintenance Abolishes the Increase in Heart Rate Seen with Liraglutide Alone: The S-LITE Randomized Trial
Authors: LUNDGREN JULIE R. , BLADBJERG ELSE MARIE , JANUS CHARLOTTE , GLIEMANN LASSE , OLSEN LISA M. , JUHL CHRISTIAN R. , JENSEN SIMON B.K. , THIRUMATHYAM ROOPAMEERA , CHRISTENSEN RASMUS M. , ANDRESEN ANNE , STALLKNECHT BENTE , HOLST JENS J. , HOVE JENS D. , MADSBAD STEN , ANTONIADES CHARALAMBOS , TOREKOV SIGNE S.
Published: June 2020

Background: Cardiovascular disease (CVD) is the leading cause of death worldwide and risk factors include obesity, physical inactivity, hypertension, and dyslipidemia. Our aim was to investigate changes in CVD risk factors after an initial weight loss followed by 1 year of treatment with glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide and/or exercise.

Method: The trial included 215 individuals with obesity (BMI 36.6 kg/m2, age 42 years). This was a double-blinded trial with an initial 8-week weight loss phase (800 kcal/day) and subsequent randomization (n=195) to placebo, placebo + exercise (150 min/week), liraglutide 3.0 mg/day or liraglutide 3.0 mg/day + exercise (150 min/week) for 1 year. Trial identifier: NCT 04122716

Results: The table shows changes in CVD risk factors after an 8-week weight loss phase followed by 1 year of treatment.

Conclusion: A 13 kg weight loss acutely improved all CVD risk factors. With liraglutide and exercise treatment combined, body weight was further reduced. Fasting glucose and HbA1c were additionally improved with the combined treatments compared to placebo and exercise alone. Interestingly, when exercise was added to liraglutide treatment there was no increase in resting heart rate as opposed to liraglutide treatment alone.

Disclosure

J.R. Lundgren: None. E. Bladbjerg: None. C. Janus: Research Support; Spouse/Partner; Mercodia, Novo Nordisk A/S. Research Support; Self; Novo Nordisk Foundation. Research Support; Spouse/Partner; Novo Nordisk Foundation. Speaker’s Bureau; Spouse/Partner; Merck Sharp & Dohme Corp. L. Gliemann: None. L.M. Olsen: None. C.R. Juhl: None. S.B.K. Jensen: None. R. Thirumathyam: Research Support; Self; Boehringer Ingelheim International GmbH. R.M. Christensen: None. A. Andresen: None. B. Stallknecht: None. J.J. Holst: Advisory Panel; Self; AstraZeneca, Merck Sharp & Dohme Corp., Novo Nordisk A/S, Zealand Pharma A/S. Other Relationship; Spouse/Partner; Antag Therapeutics. J.D. Hove: None. S. Madsbad: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Merck Sharp & Dohme Corp., Novo Nordisk A/S, Sanofi-Aventis. Research Support; Self; Boehringer Ingelheim International GmbH, Novo Nordisk A/S. Speaker’s Bureau; Self; AstraZeneca, Boehringer Ingelheim International GmbH, Novo Nordisk A/S. C. Antoniades: Board Member; Self; Caristo Diagnostics. S.S. Torekov: Research Support; Self; Novo Nordisk Inc.

Funding

Novo Nordisk Foundation (NNF16OC0019968, NNF15SA0018486); University of Copenhagen; Helsefonden; Danish Diabetes Academy
205-LB: Clinical Validation of the Implanted Prevascularized Cell Pouch as a Viable, Safe Site for Diabetes Cell Therapy
Authors: MACGILLIVARY AMANDA S. , TOLEIKIS PHILIP M. , MAZZUCA DELFINA M.
Published: June 2020

We report current clinical findings for the Cell Pouch (CP), as an implantable and retrievable medical device designed to develop a healthy vascularized tissue environment for transplantation and engraftment of therapeutic cells, which release the desired hormones for the treatment of diabetes, and other diseases. Patients with diabetes were implanted with the CP between 1-6 months, and under immunosuppression transplanted with islets isolated from donor pancreata. CPs (N=9) were explanted (between 14 -90 days post-tx). Histological sections of the explanted CPs were prepared, stained, and then reviewed by an independent pathologist for vascularization, and the presence of islets with insulin, C-peptide, and other endocrine hormones (glucagon, somatostatin) and ductal tissue (exocrine tissue) [(+) present; (++) abundant marked clusters of cells; (-) absence]. Pathology analysis revealed that 100% of the CPs were well-vascularized within a rich tissue bed. Of the CPs assessed, 89% (+ or ++) showed insulin; 78% (+ or ++) showed endocrine cells; 100% (+ or ++) showed ductal tissue. Islets were intimately related to micro-vessels in the CP tissue matrix. The amount of islets/exocrine tissue within the pre-transplant samples was similar to that found within the CPs that were explanted. With these findings, we have clinically demonstrated the vascularized Cell Pouch provides a safe and suitable, retrievable environment for the transplantation and survival of functional islets. With new emerging cellular replacement therapies as a clinical solution for all patients with diabetes, the unique Cell Pouch system solves a need to support a successful, positive clinical transplant outcome.

Disclosure

A.S. MacGillivary: Employee; Self; Sernova Corp. P.M. Toleikis: Board Member; Self; Sernova Corp. Employee; Self; Sernova Corp. Stock/Shareholder; Self; Sernova Corp. D.M. Mazzuca: Employee; Self; Sernova Corp. Stock/Shareholder; Self; Sernova Corp.
2058-P: Single-Cell Transcriptome Atlas of the Aging Mouse Pancreatic Islet Reveals Resilience to Diet-Induced Obesity
Authors: NIE JIA , MUSI NICOLAS
Published: June 2020

Aging is a key risk factor for type 2 diabetes (T2D). We examined aging mouse pancreatic islets by single-cell RNA-seq sampled across young (1 month), middle (13 months) to old (24 months) age groups. By applying single-cell Western blotting, we further verified that aged mice had increased islet beta-cell percentage, which was accompanied by an increased in vivo insulin secretion. Young and old groups were fed with a six-month high-fat diet. Age-associated enhancements in insulin secretion prevented old mice from diet-induced glucose intolerance. Together, this single-cell atlas allows a comprehensive exploration of all transcriptional states in the aging islets and provides insight into the pathogenesis of T2D. Importantly, our findings indicate that age-dependent hyperinsulinemia protects against diet-induced obesity in mice and exhibit the resilience of the islet in response to stressors.

Disclosure

J. Nie: None. N. Musi: None.

Funding

American Federation for Aging Research; San Antonio Claude D. Pepper Older Americans Independence Center (P30AG044271)
2066-P: Dyrk1a Antisense Oligonucleotides Conjugated to GLP-1R Agonist Promote Pancreatic Beta-Cell Proliferation
Authors: FU WUXIA , LEE RICHARD , PIRIE ELAINE C. , PRAKASH THAZHA , MURRAY SUE , POWERS ANDREW , MULLICK ADAM E. , CROOKE ROSANNE M.
Published: June 2020

Loss of functional beta cell mass is a major contributor to the pathogenesis of both type 1 and type 2 diabetes. Enhancing proliferation of existing beta cells in the pancreatic islet is one of the promising approaches for restoring functional beta cell mass. Increasing evidence indicates inhibition of dual specificity tyrosine-phosphorylation-regulated kinase 1A (Dyrk1a) stimulates beta cell proliferation. However, to date DYRK1a small molecule inhibitors have not been therapeutically viable due to their ability to increase proliferation in peripheral tissues. Previously we demonstrated that generation 2.5 antisense oligonucleotides (ASO) conjugated to a glucagon like peptide 1 receptor (GLP-1R) agonist can specifically target the pancreatic beta cell. When compared to control ASO treated isolated islets, islets administered a GLP-1/Dyrk1a ASO conjugate reduces DYRK1a mRNA and protein expression and significantly increases Ki67 mRNA, a cell proliferation marker, in a dose-response manner in vitro. Furthermore, combination treatment of a GLP-1/Dyrk1a ASO conjugate and the small molecule TGF-β inhibitor Ly364947, can synergistically increase Ki67 mRNA expression, cell cycle progression markers cyclin D1, cyclin E2, Cdk1, Cdk4 mRNAs and beta cell markers Pdx1 and Glut2 mRNAs when compared to a DYRK1a conjugated ASO or Ly364947 alone. Administration of a Dyrk1a conjugated ASO to wild type and db/db mice reduced Dyrk1a mRNA and increased Ki67 mRNA expression in islets when compared to a control ASO. Cell cycle markers cyclin D2, cyclin E2 and Cdk1 mRNAs are also significantly upregulated.

In conclusion, a GLP-1/Dyrk1a ASO conjugate can specifically inhibit beta cell Dyrk1a mRNA expression and promote beta cell proliferation, suggesting that this may be an important approach for the treatment of type 1 and type 2 diabetes.

Disclosure

W. Fu: Employee; Self; Ionis Pharmaceuticals, Inc. R. Lee: Employee; Self; Ionis Pharmaceuticals, Inc. E.C. Pirie: Employee; Self; Ionis Pharmaceuticals, Inc. T. Prakash: None. S. Murray: Employee; Self; Ionis Pharmaceuticals. A. Powers: Employee; Self; Ionis Pharmaceuticals, Inc. Stock/Shareholder; Self; Ionis Pharmaceuticals, Inc. A.E. Mullick: None. R.M. Crooke: Employee; Self; Ionis Pharmaceuticals, Inc. Employee; Spouse/Partner; Ionis Pharmaceuticals, Inc.
2100-P: Binding Kinetics, GLP-1 Receptor Internalization, and Effects on Insulin Secretion for GL0034 and Related GLP-1R Agonists
Authors: CARRAT GAELLE , NGUYEN-TU MARIE-SOPHIE , LECLERC ISABELLE , THENNATI RAJAMANNAR , JONES BEN , TOMAS ALEJANDRA , RUTTER GUY A.
Published: June 2020

Background and Aims: We have sought to develop novel GLP-1R agonists (RA) with improved insulinotropic effects on the pancreatic beta cell, and extended duration of action. Here, we have explored the affinity for the receptor, and action on signalling and insulin secretion, of three novel compounds.

Methods: Agonist dissociation kinetics were assessed by time-resolved FRET using SNAP-tagged GLP-1R-expressing HEK293 cells and fluorescent exendin-(9-39)-FITC. Signal bias was quantified by comparing the generation of cAMP and beta-arrestin recruitment to the GLP-1R. Recycling was measured in INS-1(832/3) cells expressing SNAP-GLP-1R, and time-lapse confocal imaging on a Zeiss LSM-780 inverted microscope. Insulin secretion was measured in vitro in islets isolated from lean C57BL6 J mice. Time-resolved FRET was used to quantify released and total hormone. Semaglutide (sema.) was supplied by Sun Pharma.

Results: GL0001, GL0028 and GL0034 showed similar binding but slower dissociation kinetics at the GLP-1R, leading to a higher binding affinity (pKd ca. 7.6) vs. semaglutide (pKd ca. 7.4). A slightly higher bias (1.3-, 1.5- and 1.5-fold, respectively, n=5) towards beta-arrestin vs. cAMP signalling was also apparent for each RA vs. sema). Coupling to cAMP generation (pEC50-pKd) was lower (1.7-, 2.0- and 1.8-fold, respectively) than sema. for each RA. Whereas endocytosis of SNAP-GLP-1R was essentially complete in 5 min. after RA (100 nM) addition for sema. and GL0034, uptake of labelled receptor was barely detectable even at 10 min. for GL0028. Efficacy in potentiating insulin secretion from isolated mouse islets at 11 mM glucose, assessed after 30 min., was similar for GL0028 and sema.

Conclusion: The novel agents GL0001 and GL0034 show similar binding properties, and impact on GLP-1R uptake, compared to sema. The more limited receptor uptake for GL0028 vs. other RA may influence the balance of central vs. pancreatic actions on insulin secretion and glucose homeostasis.

Disclosure

G. Carrat: None. M. Nguyen-Tu: None. I. Leclerc: Consultant; Spouse/Partner; Sun Pharmaceutical Industries Ltd. Research Support; Spouse/Partner; Servier. R. Thennati: None. B. Jones: None. A. Tomas: None. G.A. Rutter: Consultant; Self; Sun Pharmaceutical Industries Ltd. Research Support; Self; Servier, Sun Pharmaceutical Industries Ltd.

Funding

Medical Research Council (R022259/1); UK Wellcome Trust (212625/Z/18/Z); European Union (115881)
211-OR: Hypothalamic Slug Couples Overnutrition States to Epigenetic Reprogramming of Leptin Neural Circuits, Obesity, and Type 2 Diabetes
Authors: KIM MIN-HYUN , JIANG LIN , MYERS MARTIN G. , RUI LIANGYOU
Published: June 2020

Leptin protects against obesity and metabolic disease mainly through leptin neural circuits in the hypothalamus. Leptin binds to and activates hypothalamic leptin receptors (LepR), thereby suppressing food intake and increasing energy expenditure. Counterintuitively, obesity is associated with hyperleptinemia in rodents and humans. Moreover, leptin therapies fail to ameliorate obesity in humans. These observations support a notion that obese subjects develop leptin resistance. Multiple positive and negative regulators of LepR signaling have been identified to influence leptin sensitivity, including Sh2b1, SOCS3, and PTP1b. However, factors, which couple overnutrition states to leptin resistance, are not fully understood. Here, we observed that high fat diet (HFD) feeding markedly upregulates Slug, a nuclear epigenetic regulator, in the hypothalamus. Remarkably, LepR neuron-specific deletion of Slug (SlugΔLepR) reversed HFD-induced hyperleptinemia, energy imbalance, obesity, insulin resistance, glucose intolerance, and liver steatosis. Conversely, overexpression of Slug (MBHSlug) but not an epigenetic-defective Slug mutant in the mediobasal hypothalamus resulted in obesity, insulin resistance, glucose intolerance, and fatty liver disease in mice. We found that Slug physically binds to the LepR promoter and suppresses LepR promoter activity; accordingly, hypothalamic LepR expression was high in SlugΔLepR mice and low in MBHSlug mice. Furthermore, Slug increased the levels of LepR promoter H3K27 methylation, a repressive epigenetic mark, in the hypothalamus, presumably by recruiting H3K37 methytransferase Ezh2 to the LepR promoter. Indeed, Ezh2 inhibitor treatment blocked Slug suppression of LepR promoter activity. Collectively, we unravel a previously unrecognized Slug-elicited epigenetic reprograming of LepR pathways that couple overnutrition states to leptin resistance, obesity, and metabolic disease.

Disclosure

M. Kim: None. L. Jiang: None. M.G. Myers: Research Support; Self; AstraZeneca, Novo Nordisk Inc. L. Rui: None.

Funding

National Institutes of Health (F32DK120111)
2128-P: 12/15-Lipoxygenase Regulates Macrophage Migration during Islet Inflammation
Authors: KULKARNI ABHISHEK , PINEROS ANNIE , IBRAHIM SARA , HERNANDEZ-PEREZ MARIMAR , TERSEY SARAH A. , MIRMIRA RAGHAVENDRA , ANDERSON RYAN M.
Published: June 2020

Pancreatic islet inflammation is a shared characteristic of type 1 and type 2 diabetes pathogenesis, and macrophage infiltration has been shown to exacerbate this inflammation. The enzyme 12/15-Lipoxygenase (12/15-LOX) which is involved in polyunsaturated fatty acid metabolism, has also been implicated in islet inflammation. Based on its established roles in cellular migration in other contexts, we hypothesized that 12/15-LOX promotes macrophage infiltration. To test this hypothesis, we used two complementary model systems: zebrafish and mice. First, we knocked down 12/15-LOX in Tg(ins:NTR) zebrafish with an antisense morpholino. When these zebrafish are treated with the prodrug metronidazole (MTZ), β cells are specifically injured, and macrophages accumulate in the islet. After a 6-hour incubation with MTZ, we observed a significant reduction in the number of infiltrating macrophages in the islets of 12/15-LOX knockdown larvae as compared to control larvae (2.7±0.8 vs. 1.17±0.7, P<0.05). Similarly, injured β cells in zebrafish treated with the 12/15-LOX inhibitor ML355 showed a significant reduction in the number of islet-infiltrating macrophages compared to vehicle-treated larvae (3.67±0.7 vs. 1.2±0.3, P<0.05). Next, using transwell assays, we measured the migration of peritoneal macrophages that were isolated from 12/15-LOX knockout mice in media that was conditioned by islet culture. Again, we observed a significant reduction in macrophage migration with 12/15-LOX deficiency compared to controls (162.8±15.5 vs. 108.4±15.1, P<0.05). Mechanistically, we found that the expression of the chemokine receptor CXCR3 was significantly downregulated in 12/15-LOX-knockout macrophages. Taken together, our data reveal a role for 12/15-LOX in macrophage migration that is conserved across species. Importantly, our study indicates that the signaling pathways downstream from 12/15-LOX are potential therapeutic targets for the resolution of inflammation-induced damage in the islets.

Disclosure

A. Kulkarni: None. A. Pineros: None. S. Ibrahim: None. M. Hernandez-Perez: None. S.A. Tersey: None. R. Mirmira: Advisory Panel; Self; Hibercell, Sigilon Therapeutics, Veralox Therapeutics. Employee; Spouse/Partner; Eli Lilly and Company. R.M. Anderson: None.
2149-PUB: The Changes in the Panel of Circulating Cytokines Associated with Chronic Kidney Disease in Patients with Type 2 Diabetes
Authors: KORBUT ANTON I. , ORLOV NIKOLAI B. , FAZULLINA OLGA , KLIMONTOV VADIM
Published: June 2020

Aim: To estimate the differences in the panel of circulating cytokines mediating low-grade inflammation and fibrosis in patients with type 2 diabetes (T2D) and chronic kidney disease.Methods: We examined 130 patients with T2D duration ≥10 years. Among them, 65 individuals had reduced estimated glomerular filtration rate (eGFR; 15-59 ml/min/1.73 m2) and 64 patients had increased albuminuria. Thirty healthy subjects were acted as control. Serum IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17A, bFGF, eotaxin, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α, and VEGF were assessed by multiplex analysis. Urinary nephrin and podocin, as podocyte injury markers, and WFDC-2, a marker of tubulointerstitial fibrosis, were assessed by ELISA.

Results: Patients with T2D and reduced eGFR had increased levels of IL-1ra, IL-8, IL-13 and decreased IL-7, IL-9 and IL-15 concentrations when compared to those with preserved renal function and control (all p<0.05). Excretion of WFDC-2, but not podocyte markers, was increased in these patients (p≤0.02). The elevation of albuminuria was associated with enhanced excretion of nephrin and podocin (p<0.0001), but no differences were found in the levels of all measured cytokines between patients with normal and elevated albuminuria. The levels of IL-2, IL-4, IL-5, IL-17A, bFGF, G-CSF, IP-10 and MIP-1α correlated negatively with eGFR. No correlations were found between measured cytokines and glomerular markers. There was positive correlation between WFDC-2 and MIP-1α. The levels of IL-10 and IL-15, two anti-inflammatory cytokines, correlated negatively with WFDC-2.

Conclusion: The obtained results demonstrate that decreased eGFR and increased urinary excretion of WFDC-2, a marker of tubulointerstitial fibrosis, are associated with a predominance of pro-inflammatory and fibrogenic molecules in the panel of circulating cytokines in T2D subjects.

Disclosure

A.I. Korbut: None. O. Fazullina: None. V. Klimontov: None.
2173-PUB: Assessment of the Knowledge and Perception of Diabetic Symptoms among Patients in a Poor Environment
Authors: ADZIKA VINCENT A.
Published: June 2020

Introduction/Background: Diabetes mellitus is an endocrine disorder characterized by sustained hyperglycaemia due to loss of insulin secretion and/or impairment of insulin sensitivity at target tissues. More than 15.9 million people have diabetes accounting for more deaths in addition to infectious diseases in low income countries in Africa. Despite the alarming statistics the etiology and prognosis leading to the prevention and management of diabetes still lacks the needed attention and comprehensive education among patients in low income countries.

Aim: The aim of this study is to examine the knowledge of causes and effects of diabetic symptoms among patients in a low income country.

Method: A qualitative methodological framework focusing on participant’s illness experiences and personal beliefs about diabetes was used. We purposely recruited 30 diabetic patients who exhibited diabetic symptoms. After demographic information, each respondent completed the LANSS to ascertain neuropathy after which a semi structured interview guide was used to elicit each respondent’s view regarding diabetes. Data was transcribed and analyzed using a thematic approach.

Results: Most respondents did not know their diabetic type and the precipitating factors for their diabetic symptoms. However some reported that their diabetes was spiritual and might have been contracted through bewitchment and other supernatural means. Others traced their condition to hereditary but the main causes of diabetes were generally lacking. Majority didn’t know their medications but requested for the need to have a much potent medication.

Conclusion: Effective education and knowledge on the etiology, causes and prognosis of diabetes is generally lacking among patients living in poor countries.

Disclosure

V.A. Adzika: None.
2174-PUB: Profiling of Young Diabetes in India: A Cross-Sectional Analysis Report
Authors: NK NARAYANAN , DWARAKANATH CS , S VENKATARAMAN , RM MANIKANDAN , BS NARENDRA , DAS SAMBIT , DASH KALPANA , PALADUGU SRIDEVI , SESHADRI KRISHNA
Published: June 2020

Aim: The purpose of this study was to profile young-onset (defined as up to age 40 years) diabetes patients in India.

Methods: The current analysis is a retrospective data analysis of young-onset diabetes patients. Young age diabetes is defined as patients diagnosed with diabetes between 18 to 40 years of age. T1DM and other secondary diabetes were excluded while profiling these patients. Patient’s data was extracted from EMRs of Apollo Sugar Outpatient clinics (n=35) from January 2018 to October 2019. Descriptive statistics were applied to analyze the data and report the results.

Results: Of the total 85568 diabetes patient’s records, 4152 (5%) young diabetes patients were identified as fulfilling the criteria of T2DM of early-onset. Males were 2495 (60%) and females were 1657(40%). Mean blood pressure was 122/78 mmHg, average BMI was 28.4 kg/m2, 15% had hypertension, 39% had dyslipidemia and 1% had CAD. Positive family history was recorded in 5% of patients, and 1% were recorded to have current or past smoking. Mean HbA1c was 8.4%, FBG and PPBG were 190 mg/dL and 273mg/dL. Mean cholesterol 179mg/dL, HDL 40mg/dL, LDL 111 mg/dL, and TG 232 mg/dL and creatinine 0.9mg/dL. The average no of antidiabetes drugs prescribed was 2. Nearly 36% of the patients were on oral agents and insulins, 30% were on statins.

Conclusions: In our cross-sectional analysis 5% of patients over 18 month’s period had a diagnosis fulfilled the criteria for early-onset diabetes. There was a male preponderance. The average BMI was considerably higher. The majority had poor control and greater than a third of patients were on insulin. Other comorbidities such as hypertension and dyslipidemia were also prevalent. The higher BMI poor control and presence of comorbidities are ominous predicting a higher progression to complications given the longer disease burden in these patients. Our data indicate the need for further efforts in preventing detecting and treating diabetes in this young group of patients.

Disclosure

N. Nk: None. C. Dwarakanath: None. V. S: None. M. Rm: None. N. Bs: None. S. Das: None. K. Dash: None. S. Paladugu: None. K. Seshadri: None.
2182-PUB: Feasibility of Extended Calibration Intervals with a Long-Term, Implantable CGM System
Authors: GUPTA PIYUSH , BUTLER ANTHONY , BERTSCH KRISTA , JONES HANNAH , LEE MELISSA H. , OBEYESEKERE VARUNI R. , MARTHA SANDRA L. , JENKINS ALICIA , ONEAL DAVID N.
Published: June 2020

Background: Continuous Glucose Monitoring (CGM) technology has improved glycemia in people with diabetes. However, user burden including frequent calibration, and reinsertion of short-term devices limits adoption of this technology.

Aim: Our objective was to retrospectively analyze CGM data from a fully implanted prototype sensor to assess feasibility of an extended calibration interval.

Methods: The Eclipse® ICGM® System was surgically inserted in the lower abdomen, superficial to the rectus muscle sheath (Figure 1) in 6 adults with T1 diabetes, Mean (SD) age 42(12) years; 33% Females, HbA1c 7.1% (1.2)%/54(13) mmol/mol. For 6 months participants self-monitored blood glucose 4 times/day and wore a Dexcom G5 CGM. In addition, monthly clinic visits include meal-tests with frequent sampling YSI plasma glucose for paired analyses. Analyses of investigational device CGM data was retrospective against benchmark measurements.

Results: There were no device associated adverse events. Data from three participants demonstrate intervals of 30, 48 and 85 days between calibrations without impacting overall performance. Preliminary measures show clinically relevant accuracy of >95 % within the A+B ranges.

Conclusion: Interim data supports extended interval calibration for the fully implanted prototype Eclipse® ICGM® System which holds promise of CGM with fewer device interactions and enhanced patient experience.

Disclosure

P. Gupta: Employee; Self; GlySens Incorporated. A. Butler: Employee; Self; GlySens Incorporated. K. Bertsch: None. H. Jones: None. M.H. Lee: Research Support; Self; Medtronic. Speaker’s Bureau; Self; AstraZeneca, Medtronic. V.R. Obeyesekere: None. S.L. Martha: None. A. Jenkins: Advisory Panel; Self; Abbott, Medtronic. Research Support; Self; Abbott, GlySens Incorporated, Medtronic, Sanofi-Aventis. D.N. ONeal: None.
221-OR: Does Cell Therapy Improve the Quality of Life of Patients with Chronic Limb-Threatening Ischemia? A Randomized Controlled Trial
Authors: HUSÁKOVÁ JITKA , BEM ROBERT , NEMCOVA ANDREA , FEJFAROVA VLADIMIRA , JIRKOVSKA ALEXANDRA , DUBSKY MICHAL
Published: June 2020

Background: Autologous cell therapy (ACT) is the last option for patients with no-option chronic limb-threatening ischemia (NO-CLTI) and severe pain. The aim of our study was to assess the impact of ACT on a patient’s quality of life, chronic pain and depression using standardized questionnaires.

Methods: Forty patients with NO-CLTI were randomized and divided into groups treated initially with ACT (iACT, n=20) or standard conservative therapy (iST, n=16); 4 patients dropped out of the study. After 3 months, those in the standard therapy (ST) group were switched to (cell therapy crossover). All patients filled out the standardized EQ-5D questionnaire assessing their quality of life. Pain was evaluated using a special questionnaire including the Visual Analogue Scale (VAS) and Wong-Baker Faces (WB). The questionnaires were evaluated before randomization; in the iST group at 3 months after ST and then at 3 months after ACT.

Results: Pain was significantly reduced in the iACT group after 3 months compared to baseline in both VAS and WB scores (both p<0.001); in the iST group, we observed even worsening of both pain scores without a significant difference (NS). After the cell therapy crossover of in the iST group, both pain scores increased significantly after 3 months (both p<0.001). Quality of life measured by EQ-5D was improved after 3 months of ACT (p<0.001) while not changing in the iST group after 3 months of ST (NS). After the crossover to ACT, quality of life was significantly improved also in the iST group (p<0.001). We observed a trend in towards a decrease in depression rate after 3 months of iACT from 45 to 33%, whereas the depression rate was slightly increased after 3 months of ST (from 56 to 61%) to decrease from 56 to 18% in the iST group after cell therapy crossover (all NS).

Conclusion: Our study showed significant pain reduction in NO-CLTI patients after ACT; this treatment also led to a decrease in depression rate and overall improvement in their quality of life.

Disclosure

J. Husáková: None. R. Bem: None. A. Nemcova: None. V. Fejfarova: None. A. Jirkovska: None. M. Dubsky: None.

Funding

Ministry of Health of the Czech Republic (16-27262A); Project for Development of Research Organization (00023001); IKEM, Prague, Czech Republic
2233-PUB: Electronic Delivery Mode for Completing PROMs in the Norwegian Diabetes Register for Adults (NDR-A)
Authors: HAUGSTVEDT ANNE , HERNAR INGVILD , STRANDBERG RAGNHILD B. , COOPER JOHN G. , LØVAAS KARIANNE F. , MADSEN TONE VONHEIM , REKDAL MAGNE , IVERSEN MARJOLEIN M. , GRAUE MARIT
Published: June 2020

Background: The NDR-A is a national diabetes quality register. Norwegian health authorities specify that quality registers must include data from Patient-Reported Outcome Measures (PROMs) in addition to biomedical data. NDR-A retrieves data from electronic patient records (EPR). This raises the possibility of utilizing PROMs in clinical consultations. The main PROM included in NDR-A is the Problem Areas in Diabetes scale (PAID).

Aim: To test the feasibility of using electronic delivery mode to collect PROMs and examine how patients and health care providers experience the electronic PROMs and the EPR’s interfaces.

Methods: We invited 441 adults (>18 yrs) with type 1 diabetes to complete PROMs via 1) a web interface from their home (n=304) or 2) a kiosk (touch-screen computer) in the outpatient clinic (n=137). Using a paper questionnaire and qualitative interviews, we examined patients and health care providers experiences with the electronic PROMs and the EPR’s interfaces.

Results: Through the web interface, we reached out to 215 (71 %) of the patients of which 114 (38 %) responded. On the kiosk in the clinic, 69 (50 %) patients completed the PROMs, 24 (18 %) did not attend scheduled consultations, 5 (4 %) did not participate due to technical problems, 20 (15 %) declined participation, and 19 (14 %) did just not approach the kiosk as instructed (when a researcher was not present to remind them). Patients who completed the paper questionnaire experienced the PROMs to be relevant and were in high degree willing to complete PROMs annually. In the qualitative interviews, health care providers reported the PROMs and the EPR’s interfaces to be appropriate to improve the focus on patients’ diabetes-related psychological and emotional challenges and needs.

Conclusion: The tested modes for electronic capture of PROMs have limitations in reaching patients. However, both patients who completed the PROMs and the health care providers were satisfied with the electronic PROMs and the EPR’s interfaces.

Disclosure

A. Haugstvedt: None. I. Hernar: None. R.B. Strandberg: None. J.G. Cooper: None. K.F. Løvaas: None. T. Madsen: None. M. Rekdal: Employee; Self; DIPS AS. M.M. Iversen: None. M. Graue: None.