bluuebunny/crossref_metadata_2025_split huggingface dataset:
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&lt;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&lt;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&lt;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&lt;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&lt;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&lt;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)