bluuebunny/crossref_metadata_2025_split huggingface dataset:
474-P: Apabetalone (RVX-208) Attenuates an Inflammatory Milieu that Enhances Adhesion of Monocytes to Endothelial Cells in Type 2 Diabetes Mellitus with Cardiovascular Disease Patients
Authors: TSUJIKAWA LAURA , KULIKOWSKI EWELINA , CALOSING CYRUS , WASIAK SYLWIA , GILHAM DEAN , HALLIDAY CHRISTOPHER , JOHANSSON JAN O. , SWEENEY MICHAEL , WONG NORMAN C. 
Published: June 2019

To explore mechanisms underlying a 57% relative risk reduction of major adverse cardiovascular events (MACE) in type 2 diabetes mellitus (T2DM) patients (pts) with CVD given 200 mg apabetalone (APL) by mouth. APL is a small molecule that inhibits bromodomain and extra-terminal (BET) proteins, which are epigenetic readers of acetylated lysine in histones contained within actively transcribing chromatin. Inhibiting BET proteins attenuates transcribed genes that are upregulated in disease states. Studies here used SOMAscan proteomics of patient plasma given placebo (n=30) or APL (n=25) and cultured monocyte (THP-1) or endothelial (HUVEC) cells. Results showed differences in the proteome spanning 4 CVD pathways: acute phase response, intrinsic prothrombin activation, leukocyte extravasation signaling, and coagulation. This data showed TNFα target genes were preferentially upregulated in T2DM+CVD (p&lt;0.001; z-score = 2.270) vs. euglycemic pts, suggesting an inflammatory state that was attenuated by APL (p&lt;0.001; z-score = -2.308). To mimic inflammation in vitro, TNFα (10 ng/ml) added to co-culture of THP-1 and HUVEC induced adhesion 12-fold in euglycemia; APL inhibited it by 44%. HUVECs exposed to TNFα induced IL-1β, E-selectin, VCAM1 and IL-6 mRNA by 1685, 1164, 196 and 9-fold respectively while APL inhibited (50-99%). In THP-1, TNFα induced IL-1β, MYD88 and CD-44 mRNA by 3.5, 2.6, and 1.8-fold respectively; inhibited by APL (39-84%). To mimic T2DM, high glucose (HG, 25.6 mM) induced adhesion of THP-1 to HUVEC by 2.4-fold was blocked by APL. In HG, Very Late Antigen-4 (VLA-4) mRNA, a THP-1 adhesion gene, rose 1.3-fold; APL blocked it &gt;50%. HG induced adhesion genes in HUVECs, E-selectin and MYD88, by 2- and 1.3-fold was lowered by APL.
In summary, APL lowers MACE in T2DM+CVD pts by attenuating expression of genes that mediate adhesion of monocytes to endothelial cells. This hypothesis is being tested in a phase 3 trial BETonMACE.

Disclosure
L. Tsujikawa: None. E. Kulikowski: Employee; Self; Resverlogix Corp. Stock/Shareholder; Self; Resverlogix Corp. C. Calosing: None. S. Wasiak: Employee; Self; Resverlogix Corp. Stock/Shareholder; Self; Resverlogix Corp. D. Gilham: Employee; Self; Resverlogix Corp. C. Halliday: Employee; Self; Resverlogix Corp. Stock/Shareholder; Self; Resverlogix Corp. J.O. Johansson: Employee; Self; Resverlogix Corp. M. Sweeney: Employee; Self; Resverlogix Corp. N.C. Wong: Employee; Self; Resverlogix Corp. Stock/Shareholder; Self; Resverlogix Corp.
48-OR: Gut Dysbiosis Promotes Diabetic Retinopathy (DR) through TLR-2 Activation by Peptidoglycan (PGN) in Angiotensin Converting Enzyme 2 (ACE2) Deficient Type 1 Diabetic (T1D) Mice
Authors: PRASAD RAM , DUAN YAQIAN , FLOYD JASON L. , GRANT MARIA B. 
Published: June 2019

Purpose: Previously, we demonstrated that ACE2 deficiency increases the severity of DR in Akita mice. In this study, we sought to determine the mechanism where by the gut microbiome, which is typically altered in T1D, contributes to the pathogenesis of DR in ACE2-/-Akita mice.
Methods: Wild type (WT), Akita and ACE2-/-Akita mice were studied at 9 months of diabetes and the gut microbiota was examined using 16S rRNA sequencing and metatranscriptomic analysis. Microbial load in the circulation was assessed by measuring circulating PGN levels along with markers of the gut vascular barrier (GVB). Next, human retinal endothelial cells (HREC) were treated with PGN, a known Toll-like receptor (TLR)-2 agonist, to determine whether PGN enhances permeability.
Results: 16S rRNA sequencing revealed a shift in beta diversity of the gut microbiota with an increase in Firmicutes and Bacteroidetes within the gut of ACE2-/-Akita mice. Metatranscriptomic analysis showed increases in PGN synthesis. Loss of ACE2 promoted gut barrier permeability through decreasing the turnover of VE-cadherin expression (41%, p&lt;0.0001) in the intestine of ACE2-/-Akita mice. Increased vascular permeability was associated with barrier dysfunction, microbial translocation and increase PGN in the circulation (2.5 fold, p&lt;0.007) in ACE2-/-Akita mice compared with Akita mice. Acellular capillaries (2 fold, p&lt;0.05) were significantly higher in ACE2-/-Akita mice compared with Akita mice. PGN treatment caused dissociation of VE-cadherin junctional complexes in HRECs through a MyD88/ARNO/ARF6-mediated signaling pathway. This occurred by depleting p120-catenin (51%. p&lt;0.004) and internalization of VE-cadherin.
Conclusions: These data suggest that microbiota-derived PGN has deleterious effects on retinal endothelial cells to increasing permeability. Strategies aimed at resorting gut health may serve to prevent development of DR.

Disclosure
R. Prasad: None. Y. Duan: None. J.L. Floyd: None. M.B. Grant: None.

Funding
National Eye Institute (EY028858)
483-P: Pioglitazone Suppresses Macrophage Proliferation in Apolipoprotein-e Deficient Mice by Activating PPARγ
Authors: MURAKAMI-NISHIDA SAIKO , MATSUMURA TAKESHI , NISHIDA SHUHEI , SENOKUCHI TAKAFUMI , ISHII NORIO , YAMADA SARIE , MORITA YUTARO , WADA TOSHIAKI , MOTOSHIMA HIROYUKI , KONDO TATSUYA , ARAKI EIICHI 
Published: June 2019

Background and Aims: Local macrophage proliferation in the atherosclerotic plaque is linked to the enhanced atherosclerosis progression. Although we reported that troglitazone, a thiazolidinedione (TZD), suppressed oxidized low-density lipoprotein (Ox-LDL)-induced macrophage proliferation, its mechanisms have been unclear. We therefore investigated the mechanism(s) of the inhibitory effect of macrophage proliferation by another TZD, pioglitazone.
Methods: High-fat diet (HFD)-fed, apolipoprotein E-deficient (ApoE KO) mice were treated orally with pioglitazone (10 mg/kg/day). After 8 weeks of treatment, the whole aorta or frozen sections of the aortic sinus were stained with Oil red O. To identify macrophage proliferation and apoptosis, we performed Ki67/Iba-1 double immunohistochemical staining and TUNEL method, respectively.
Results: Atherosclerosis progression was suppressed in aortic sinuses of pioglitazone-treated ApoE KO mice, and they showed fewer proliferating macrophages in the plaques. Pioglitazone suppressed Ox-LDL-induced macrophage proliferation in a dose-dependent manner, which was ameliorated by peroxisome proliferator-activated receptor-γ (PPARγ) siRNA. Low concentrations (less than 100 µM) of pioglitazone, which can suppress macrophage proliferation, activated PPARγ in macrophages, but did not induce macrophage apoptosis. Pioglitazone did not induce TUNEL-positive cells in atherosclerotic plaques in ApoE KO mice.
Conclusions: Pioglitazone suppresses macrophage proliferation through PPARγ without inducing macrophage apoptosis. These findings imply that pioglitazone could prevent macrovascular complications in diabetics.

Disclosure
S. Murakami-Nishida: None. T. Matsumura: None. S. Nishida: None. T. Senokuchi: None. N. Ishii: None. S. Yamada: None. Y. Morita: None. T. Wada: None. H. Motoshima: None. T. Kondo: None. E. Araki: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Novo Nordisk Inc., Takeda Pharmaceutical Company Limited. Speaker's Bureau; Self; Astellas Pharma Inc., Merck & Co., Inc., Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi.
484-P: Acute Hyperglycemia In Vivo Promotes a Proatherogenic Monocyte Phenotype
Authors: ZHANG RONG MEI , OH JISU , LIN XIAOBO , STONE PAMELA , WHARTON KATHLEEN J. , ARBELAEZ ANA MARIA , BERNAL-MIZRACHI CARLOS 
Published: June 2019

Diabetes is a chronic inflammatory disease that accelerate cardiovascular disease (CVD). However, the contribution of hyperglycemia to inflammation and CVD risk is unclear. Animal studies have shown that monocytes carry cholesterol to the atherosclerotic plaque. Thus, it is key to determine if hyperglycemia can exert a pro-atherogenic effect in human monocytes in vivo. We performed octreotide euglycemic-hyperglycemic clamps in 21 healthy subjects, 20-45 yo. Monocytes isolated during euglycemia (90 mg/dl) and after 2h of hyperglycemia (300 mg/dl) were evaluated for cholesterol metabolism and monocyte function (adhesion to fibronectin, transwell migration assays). Also, we assessed the effects of hyperglycemia on monocyte cholesterol metabolism and function from 10 healthy controls cultured in euglycemia or high glucose media with or without 2-deoxyglucose (2DG) at 2 and 6 hours. Acute hyperglycemia in vivo increased 37% total monocyte cholesterol content (TC) (p&lt;0.02), 40% cholesterol ester (CE)(p&lt;0.02) and upregulated oxidized low-density lipoprotein (oxLDL) cholesterol uptake by by 24%(p&lt;0.01). Moreover, acute hyperglycemia increased monocyte adhesion and transwell migration (p&lt;0.01) suggesting cholesterol full monocytes carry cholesterol into the vessel wall. In vitro studies confirmed that hyperglycemia increased monocyte adhesion and migration by 20% and monocyte TC by 20% and CE by 40% at 2 and 6 hours (p&lt;0.03) by increasing oxLDL or LDL cholesterol uptake. Hyperglycemia didn’t affect monocyte cholesterol biosynthesis or cholesterol efflux induced by HDL or APOA1. Addition of 2DG cultured monocytes in both glucose conditions prevented hyperglycemia induced monocyte adhesion migration and monocyte CE and TC content at 2 and 6 hours. Thus, acute hyperglycemia in vivo without hyperinsulinemia induce monocyte glycolysis increasing monocyte cholesterol uptake, total cholesterol content it activates monocytes adhesion and migration, all key components to accelerate atherosclerosis.

Disclosure
R.M. Zhang: None. J. Oh: None. X. Lin: None. P. Stone: None. K.J. Wharton: None. A. Arbelaez: None. C. Bernal-Mizrachi: None.

Funding
National Heart, Lung, and Blood Institute (R01HL094818-06); National Institute of Neurological Disorders and Stroke (P01NS080675)
487-P: The Activation of mTORC1 in Leukocytes of Patients with Cancer and Diabetes
Authors: VATSEBA TAMARA , SOKOLOVA LIUBOV K. , PUSHKAREV VIKTOR V. , KOVZUN OLENA I. , PUSHKAREV VOLODYMIR M. , TRONKO MYKOLA D. 
Published: June 2019

Aim: mTOR/p70S6K1 play an important role in the pathogenesis of cancer and diabetes. Macrophages and lymphocytes are involved in the pathogenesis of diabetes, diabetic atherosclerosis, formation of insulin resistance as well as immune response to cancer and tumor maintenance. Thus, the aim was to study the mTORC1 activation by determining the phosphorylation of PRAS40 and p70S6K1 in leukocytes of patients with type 2 diabetes and cancer.
Materials and Methods: The following groups were studied: control group - healthy people, representative by age (n=6), patients with type 2 diabetes (n=12), cancer patients (n=15) and patients with both cancer and diabetes (n=7). For determinations of phospho-PRAS40 (Thr246) and phospho-p70S6K1 (Thr389) amounts ELISA kits КНО0421 and 85-86053 (Invitrogen) respectively were used. The protein concentration in the lysate was determined using BCA protein assay kit. The measurements were carried out on a microplate reader at a wavelength of 450 nm.
Results: The amounts of phosphorylated PRAS40 and p70S6K1 significantly increase in leukocytes of patients with type 2 diabetes and cancer, which indicates the mTORC1 activation. The number of phospho-PRAS40-positive samples in patients with diabetes was 83.3%, and in patients with cancer - 66.7%. The decrease in the amount of phospho-PRAS40 in leukocytes of the patients group both with cancer and diabetes was observed.
Conclusions: Thus, phosphorylation of PRAS40 is generally consistent with the activation of mTORC1, as measured by phosphorylation of p70S6K1. The changes in the activity of mTOR/S6K in severe chronic diseases can be systemic, and the study of the status of these kinases in leukocytes perhaps could be useful for diagnostic and prognostic purposes.

Disclosure
T. Vatseba: None. L.K. Sokolova: None. V.V. Pushkarev: None. O.I. Kovzun: None. V.M. Pushkarev: None. M.D. Tronko: None.
488-P: Effects of Novel Antidiabetic Therapy on Platelet Reactivity in Patients with Type 2 Diabetes Mellitus
Authors: SIASOS GERASIMOS , BLETSA EVANTHIA , STAMPOULOGLOU PANAGIOTA K. , BATZIAS KONSTANTINOS , ANTONOPOULOS ALEXIOS , PASCHOU STAVROULA A. , TSIGKOU VASILIKI , GOULIOPOULOS NIKOLAOS , MAZARIS SAVVAS , VOGIATZI GEORGIA , TSALAMANDRIS SOTIRIS , OIKONOMOU EVANGELOS , ZAROMYTIDOU MARINA , POLITOU MARIANNA , VRYONIDOU ANDROMACHI , THANOPOULOU ANASTASIA , TENTOLOURIS NIKOLAS , TOUSOULIS DIMITRIS 
Published: June 2019

Background: Patients with type 2 diabetes mellitus (DM2) exert a higher risk for thrombotic events. Platelet reactivity may be used to assess prothrombotic state in patients with cardiovascular disease. We aimed to investigate whether the administration of novel antiglycemic agents may influence platelet reactivity in these patients.
Methods: We enrolled 99 patients (male=63.3%) receiving metformin for DM2 who did not achieve therapeutic targets under current treatment. Subjects were assigned to age and sex matched groups (n=33 per group) of an additional antiglycemic agent; either DPP-4 inhibitor, SGLT2 inhibitor or a GLP-1 agonist. Platelet reactivity was measured with PFA-200 assay; collagen/epinephrine (CEPI) and collagen/ADP closure time (CADP) were calculated in seconds and HbA1c at baseline and at 3 months.
Results: There was no difference for male gender (p=0.10) or age (64.92 ± 8.30 years, p=0.27) between the study groups. All patients achieved better glycemic control in terms of HbA1c values between baseline and follow-up (7.78% vs. 6.92% for DPP-4, 7.52% vs. 6.73% for SGLT2 and 8.19% vs. 6.85% for GLP-1, p&lt;0.001 for all). Platelet reactivity did not differ significantly at baseline between the study groups (p=0.71 for CEPI and p=0.6 for CADP). Additionally, CEPI (p=0.54) and CADP (p=0.43) did not change significantly at follow-up in all groups. Interestingly, when we conducted a separate analysis for patients not receiving antiplatelet agents, we observed that only CADP was significantly reduced at follow-up time after DPP-4 and GLP-1 treatment, but not after SGLT2 administration (148.50 ± 60.65 vs. 119.75 sec for DPP-4, p=0.01, 104.12 ± 25.68 vs.88.7 ± 22.25 sec for GLP-1, p=0.01 and 101.57 ± 15.26 vs. 88.57 ± 13.26 sec for SGLT2, p=0.73).
Conclusion: The addition of DPP-4 or GLP-1 to metformin influences platelet reactivity and achieves better glycemic control in DM2 patients, providing further insights in the pathophysiology of DM2.

Disclosure
G. Siasos: None. E. Bletsa: None. P.K. Stampouloglou: None. K. Batzias: None. A. Antonopoulos: None. S.A. Paschou: None. V. Tsigkou: None. N. Gouliopoulos: None. S. Mazaris: None. G. Vogiatzi: None. S. Tsalamandris: None. E. Oikonomou: None. M. Zaromytidou: None. M. Politou: None. A. Vryonidou: None. A. Thanopoulou: None. N. Tentolouris: None. D. Tousoulis: None.
491-P: Obesity-Induced Kidney Damage Is Attenuated by Genetic Deletion of miR-379 in Mice
Authors: ABDOLLAHI MARYAM , KATO MITSUO , TUNDUGURU RAGADEEPTHI , LANTING LINDA L. , NATARAJAN RAMA 
Published: June 2019

Obesity is significantly associated with diabetes and related complications like kidney disease. Evidence shows dysregulation of certain non-coding RNAs including microRNAs can accelerate diabetic kidney disease (DKD). Recently, we found that miR-379 and its host transcript long non-coding RNA, lncMGC, were regulated by endoplasmic reticulum (ER) stress in glomerular cells, and lncMGC could promote features of DKD in mice. In this study, we examined the in vivo role of miR-379 in obesity-induced kidney disease by testing miR-379-knockout (KO) mice generated by CRISPR-Cas9 editing. Male (M) and female (F) mice were fed with control diet, or high fat diet (HFD) for 24 weeks to induce obesity in wild type (WT) and miR-379KO mice. Kidney function was examined by measuring urinary albumin and creatinine. Expression of miR-379 target genes (Edem3, Fis1), Leptin receptor (LepR) and profibrotic genes (Tgf-β1, Ctgf, Col4a1, Col1a2) was measured in isolated glomerular cells. Cortical sections were stained for histopathology. Blood glucose levels were similar in miR379KO-HFD and WT-HFD. miR379KO-HFD-F mice exhibited significantly lower body weight and body fat composition versus WT-HFD-F. Hyperinsulinemia observed in WT-HFD-F mice was attenuated in miR379KO-HFD-F. LepR expression was significantly decreased in WT-HFD-F but not miR379KO-HFD-F mice relative to controls. Increased expression of profibrotic genes, glomerular hypertrophy and interstitial fibrosis observed in WT-HFD-F were significantly attenuated in miR379KO-HFD-F mice. Similar to females, male miR-379KO-HFD-M mice showed significant reductions in glomerular hypertrophy, fibrosis and ER stress markers vs. WT-HFD-M. However, unlike females, miR379KO-HFD-M mice did not display reduced body weight compared to WT-HFD-M. These results suggest miR-379 deficiency protects mice from HFD-induced obesity and kidney injury, with some gender differences. miR-379 inhibition may be a novel therapy for HFD-induced metabolic disorders.

Disclosure
M. Abdollahi: None. M. Kato: None. R. Tunduguru: None. L.L. Lanting: None. R. Natarajan: None.

Funding
National Institutes of Health
507-P: Influence of Past Information to Precision of Diabetic Nephropathy Aggravation Prediction
Authors: KOSEKI AKIRA , ONO MASAKI , KUDO MICHIHARU , HAIDA KYOICHI , MAKINO MASAKI , SUZUKI ATSUSHI 
Published: June 2019

Background: In the presence of large data set of electronic health records (EHRs), predicting the future disease status is of importance for decision making in the medical treatments. Using modern machine learning techniques, it is generally becoming easier to build complex models to predict the future. For those models, a set of past information are used to make explanatory variables, however, we don't have enough knowledge as to how long we should collect data backward. In some cases, very late tendencies are influencing the future status of disease while in the other cases, old events were the importance causes of the change of the disease status. Our interest thus lies in how old data we have to process to make the good prediction models.
Method: In this paper, we discuss a set of machine learning algorithms to predict the diabetic nephropathy stage in the future using sets of input variables which were collected from different time span of past records. To compare the performance of algorithms we used Logistic Regression, AdaBoost, Gradient Boosting, Decision tree, Multi-layer Perceptron, and Random Forest. We then provide different set of variables of EHR that include past 30-, 60-, 90-, 180-, 210-, 240-, 270-, 300-, 330-, and 360-day data sets, from which we extracted several longitudinal statistics for input variables. From about 65 thousand type 2 diabetes patients, the models classify whether the nephropathy stage gets aggravated or stay in 180 days.
Results: For almost all algorithms, AUC is getting improved when using older data, and 360-day data sets gave the best. Among the algorithms, Gradient Boosting gave the best AUC of 0.77 when using 360-day data set. When using 360-day data sets, Decision Tree gave worst AUC of 0.61.
Conclusion: We observed that when using to past data up to 360 days, the oldest data set gave the best prediction performance. Longitudinal statistics in rather long span gives good explanatory information for future nephropathy development.

Disclosure
A. Koseki: Employee; Self; IBM. M. Ono: None. M. Kudo: Employee; Self; IBM. K. Haida: None. M. Makino: None. A. Suzuki: Research Support; Self; Chugai Pharmaceutical Co., Ltd., Dai-ichi Life Insurance Company, IBM, MSD, Ono Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker's Bureau; Self; Asahi Kasei Corporation, Daiichi Sankyo Company, Limited, Eli Lilly and Company, Mitsubishi Tanabe Pharma Corporation, Taisho Pharmaceutical Co., Ltd.
511-P: The Renoprotective Effect of Evogliptin, a Dipeptidyl Peptidase-4 Inhibitor, in a Mouse Model of Renal Fibrosis
Authors: KIM NAYOUNG , LEE IN-KYU , PARK SUNG DON , KIM MIN-JI , KIM JUNG-GUK , CHOI YEON-KYUNG , PARK KEUN-GYU , KIM MI-JIN 
Published: June 2019

Background: Renal tubulointerstitial fibrosis is a major pathologic finding of nearly all cause of end-stage renal disease. Dipeptidyl peptidase-4 (DPP-4) inhibitors are widely used in the treatment of patients with type 2 diabetes and have proven protective effects on diabetic kidney disease. This study aimed to investigate possible beneficial effects of evogliptin on renal fibrosis in mice.
Methods: We examined the effects of evogliptin on renal tubulointerstitial fibrosis following unilateral ureteral obstruction (UUO) in mice. We further defined the role of evogliptin on transforming growth factor-β (TGF-β)/Smad signaling pathways in renal tubulointerstitial fibrosis through in vivo and in vitro study.
Results: Through hematoxylin/eosin and sirius red staining, we observed that evogliptin effectively ameliorates UUO-induced renal atrophy and fibrosis. Immunohistochemistry in conjunction with western blot analysis revealed that evogliptin treatment inhibited UUO-induced up-regulation of phosphorylated Smad3, α-smooth muscle actin, plasminogen activator inhibitor 1, fibronenctin, and type 1 collagen. In vitro experiments with proximal tubule epithelial cell suggested that the effects of evogliptin on UUO-induced renal fibrosis are mediated by inhibition of the TGF-β/Smad signaling pathway.
Conclusion: The present study demonstrates that evogliptin has a protective effect on UUO-induced renal tubulointerstitial fibrosis, suggesting that its clinical applications could extend to the treatment of nondiabetic origin renal disease.

Disclosure
N. Kim: None. I. Lee: Board Member; Self; NovMetapharm Co, Inc. S. Park: None. M. Kim: None. J. Kim: None. Y. Choi: None. K. Park: None. M. Kim: None.

Funding
National Research Foundation of Korea