bluuebunny/crossref_metadata_2025_split huggingface dataset:
462-P: AT-001, a Next Generation Aldose Reductase Inhibitor with Improved Selectivity and Specificity, Protects from Cellular Damage Associated with Hyperglycemia
Authors: PERFETTI RICCARDO , YEPURI GAUTHAM , QUADRI NOSIRUDEEN A. , RAMASAMY RAVICHANDRAN , GHANNAM AMEEN F. , SHENDELMAN SHOSHANA 
Published: June 2020

The generation of Reactive Oxygen Species (ROS) and Advanced Glycation Endproducts (AGE’s) resulting from chronic tissue exposure to elevated levels of glucose has been identified as a key modulator of diabetic complications including diabetic cardiomyopathy. Abnormal activation of the polyol pathway converts excess glucose to sorbitol, generating ROS and AGEs. This conversion is catalyzed by the enzyme Aldose Reductase. Inhibition of Aldose Reductase, the rate limiting enzyme in the polyol pathway, reduces the production of sorbitol, attenuates increases in NADH, and down-regulates the synthesis of ROS.
Methods and Results: Cultured human adult cells (NHK) were exposed to elevated glucose [25mM] to simulate hyperglycemic conditions in diabetic tissue in the presence/absence of Aldose Reductase inhibitor AT-001 [0.18nM]. AT-001 dose selection was based on previous dose response studies. Cytosolic oxidative stress was evaluated and quantified using dihydroethidium (DHE) staining and quantitated via colorimetric assessment. Mitochondrial-specific oxidative stress ROS levels were quantitated using MitoSOX staining. AT-001 prevented the production and accumulation of ROS as assessed by both DHE quantitation and MitoSOX staining, demonstrating effective inhibition of polyol pathway associated damage to these cells.
Conclusions: AT-001 effectively prevented cellular damage caused by oxidative stress under hyperglycemic conditions warranting further investigation of AT-001 in the treatment of diabetic complications. AT-001 is currently being evaluated in a pivotal phase 2/3 study (ARISE-HF NCT 04083339) to treat diabetic cardiomyopathy and prevent progression to overt heart failure and also includes sub-analyses for retinopathy and diabetic peripheral neuropathy.

Disclosure
R. Perfetti: Stock/Shareholder; Self; Applied Therapeutics. G. Yepuri: None. N.A. Quadri: None. R. Ramasamy: Consultant; Self; Applied Therapeutics. A.F. Ghannam: Employee; Self; Applied Therapeutics Inc. Stock/Shareholder; Self; Sanofi US. S. Shendelman: Employee; Self; Applied Therapeutics. Stock/Shareholder; Self; Applied Therapeutics.

Funding
Applied Therapeutics (NCT04083339)
467-P: Brown Fat Original Mir-30b Targeting Renal RUNX1 to Improve Diabetic Nephropathy
Authors: GUAN MEIPING 
Published: June 2020

It has been verified that adipose tissue was the major source of circulating microRNAs (miRNA), which could target downstream genes in distant organs such as liver. Here we show that brown adipose tissue (BAT) transplantation from healthy mice donor could reactivate endogenous BAT activity of diabetic mice induced by high fat diet (HFD) and streptozotocin (STZ) injection. BAT activation increased circulating miR-30b level and significantly improved diabetic nephropathy. In vitro, miR-30b was significantly downregulated by TGF β1 in HK-2 cells. In vivo, agomiR-30b was directly administered to the BAT of diabetic mice (0.25nmol/g body weight) twice per week for 4 consecutive weeks. Over-expressing miR-30b markedly decreased fibronectin expression, coupled with downregulated Runt-related transcription factor 1(Runx1) and snail family zinc finger 1 (Snail1) in the kidney of diabetic mice, while silencing of miR-30b displayed the opposite features. Next, knockdown or overpression of Runx1 both in vitro by using siRUNX1 or pCMV-RUNX1, and in vivo by AAV-U6-shRunx1 or AAV-EF1a-Runx1 respectively, could mimic the above phenotype of miR-30b mimic and inhibitor treatment respectively. Moreover, Runx1 promoted TGFβ1-induced fibrosis through upregulating PI3K pathway. These results suggest that BAT transplantation ameliorates diabetic nephropathy in mice through reactivating endogenous BAT and increasing circulating miR-30b level, which subsequently target renal Runx1 and Snail1, to improve renal fibrosis. BAT-original miRNAs might be a promising target for kidney protection in diabetes mellitus.

Disclosure
M. Guan: None.

Funding
National Natural Science Foundation of China (81870612, 81628004, 31400992, 81470047); Guangdong Science and Technology Project (2019A1515011997)
472-P: The Profiles of MicroRNAs from Urinary Extracellular Vesicles (EVS) Prepared by Various Isolation Methods and Their Correlation with Serum EV MicroRNAs in Patients with Type 2 Diabetes and Diabetic Nephropathy
Authors: KIM YOOMI , LEE DAEHO , LEE KIYOUNG 
Published: June 2020

MicroRNAs that circulate in biological fluids are frequently enclosed in extracellular vesicles. However, urinary EVs and their cargo miRNAs have not been systematically studied according to their EV isolation methods. In type 2 diabetics with nephropathy (n=4), we compared miRNA species in urine EVs prepared by various isolation methods, including ultracentrifugation, qEV original size exclusion column, ExoQuick-TC Plus (ExoQuick), and simple ultrafiltration using Amicon Ultra centrifugal filter devices (Amicons) 10K and 100K. EV miRNAs were profiled by next-generation sequencing . We evaluated the correlations of EV miRNA expression between the urine and serum samples isolated by UC. From each of 100 ml of urine, the UC method yielded the highest number of EV miRNA species (233 ± 37.3), with the ExoQuick yielded the lowest (103 ± 17.4). Urine EV miRNA profiles were highly correlated between UC, qEV, ExoQuick and Amicon 10K methods. EV miRNA profiles between the urine and serum samples showed variable correlations between the patients (paired sample number=3, r =0.39-0.72). we examined EV miRNAs from urine isolated using two pore sizes with 10 kDa and 100 kDa filters. The expression level patterns of EV miRNAs isolated from urine were strongly correlated between Amicon 10K and the other three validated methods (UC, qEV, and ExoQuick). Pearson correlation coefficients between Amicon 100K and the three methods varied from 0.19 to 0.33. UC, qEV, ExoQuick, and Amicon 10K are suitable for urinary EV isolation to profile miRNAs. Urine- and serum-derived EV miRNA profiles have positive and variable correlations depending on specific patients. Further studies are required to identify a factor that influences these correlations and to determine the sample type that better describes the clinical and pathophysiological status of diabetic patients with diabetic kidney disease.

Disclosure
Y. Kim: None. D. Lee: None. K. Lee: None.
478-P: Nox5 Aggravates Renal Injury Independent of Nox4 Expression in Diabetes
Authors: JHA JAY C. , URNER SOFIA , DAI AOZHI , COOPER MARK E. , RODEN MICHAEL , KENNEDY CHRIS , JANDELEIT-DAHM KARIN 
Published: June 2020

Background: Oxidative stress plays an important role in the pathogenesis of diabetic kidney disease (DKD). Evidences suggest roles for pro-oxidant enzymes Nox4 and Nox5 in animal models of DKD. Nox5 is present in mans but not in mice/rats. It appears that Nox5 could be a main culprit in human DKD. Therefore, we aimed to examine the roles of Nox5 vs. Nox4 and their relative contribution to renal pathology in DKD.
Methods: We examined the expression and interaction of Nox5 and Nox4 and ROS formation in human kidney biopsies. In vitro, certain human renal cells being knockdown for Nox4 and Nox5 were exposed to high glucose. In vivo, we examined the effect of Nox5 expression in the absence of Nox4 in STZ- diabetic mice. We developed rabbit models of DKD and Nox5KO rabbits.
Results: Increased Nox5 expression and enhanced ROS level was seen in human diabetic kidney biopsies compared to nondiabetic kidney. Nox5 shows the highest upregulation in human renal cells exposed to high glucose in comparison to other Nox isoforms. Nox5 silencing attenuated high glucose induced increased expression of markers of fibrosis, inflammation and putative elements via reduction in ROS formation. Nox5 appears to be upstream of Nox4 and that Nox5 inhibition downregulates Nox4, but not vice versa. In vivo, cell specific expression of Nox5 in both Nox4 KO and GKT137831 (a renal Nox4 inhibitor) treated diabetic mice demonstrated a 30-40% increase in albuminuria, mesangial expansion, renal fibrosis and inflammation as well as enhanced ROS production in comparison to diabetic mice not expressing Nox5. In addition, both high fat feeding and alloxan induced diabetic rabbits showed increased renal Nox5 expression in association with increased renal injury along with upregulation of CTGF, fibronectin and MCP-1 as well as enhanced renal ROS production.
Conclusions: These findings provide evidence that Nox5 plays a superior pathogenic role in DKD in comparison to Nox4. Therefore, targeting Nox5 may represent a better approach to treat and prevent DKD in human.

Disclosure
J.C. Jha: None. S. Urner: None. A. Dai: None. M.E. Cooper: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim International GmbH, Mundipharma International. Research Support; Self; Boehringer Ingelheim International GmbH, Novo Nordisk A/S. Speaker’s Bureau; Self; AstraZeneca, Merck Sharp & Dohme Corp., Novartis AG, Sanofi-Aventis, Servier. M. Roden: Advisory Panel; Self; Servier. Board Member; Self; Poxel SA. Consultant; Self; Eli Lilly and Company, Gilead Sciences, Inc., ProSciento, TARGET PharmaSolutions. Research Support; Self; Boehringer Ingelheim International GmbH, Novartis Pharma K.K., Sanofi US. Speaker’s Bureau; Self; Novo Nordisk A/S. C. Kennedy: None. K. Jandeleit-Dahm: None.
481-P: Renal Distribution and Expression Analysis of ROCK Isoforms in Diabetic Kidney Disease
Authors: MATOBA KEIICHIRO , TAKEDA YUSUKE , NAGAI YOSUKE , AKAMINE TOMOYO , KANAZAWA YASUSHI , KAWANAMI DAIJI , UTSUNOMIYA KAZUNORI , NISHIMURA RIMEI 
Published: June 2020

The small GTPase Rho and its effector, Rho-kinase (ROCK), are implicated in the fundamental cellular processes (e.g., cell adhesion, migration). Studies over the past decade have illuminated ROCK as an important regulator of diabetic kidney disease. We previously demonstrated elevated renal ROCK activity in both type 1 and type 2 diabetic murine models. In an attempt to elucidate the roles of the activated ROCK, we provided evidence that ROCK inhibition attenuates an increase of urinary albumin excretion, glomerular sclerosis, and podocyte loss in rodents. Mechanistically, ROCK regulates inflammatory events as well as hypoxia-induced fibrotic reactions. However, specific roles of ROCK isoforms, ROCK1 and ROCK2, in diabetic kidneys remain unclear. In the present study, we first investigated the renal distribution of ROCK1 and ROCK2 using wild type mice and human kidneys obtained at necropsy. The immunohistochemical analysis detected both isoforms in glomeruli, tubules, and vessel walls. Whereas ROCK1 was mainly expressed in the cytoplasm, ROCK2 was accumulated in the nucleus of these cells. Double staining with anti-nephrin antibody showed co-localization of ROCK2 with nephrin, indicating distinctive roles of ROCK2 in the glomerular podocyte. In order to elucidate the role of podocyte ROCK2, we next evaluated gene expression levels of ROCK2 in high-fat diet-fed mice and patients with diabetic kidney disease. In both mice and patients, ROCK2 was increased in the setting of diabetes compared to healthy subjects. Of note, glomerular ROCK2 mRNA levels were positively correlated with the increase of urinary albumin excretion in mice. When considered alongside our previous observations, the current work highlights the importance of ROCK2 in diabetic podocytopathy.

Disclosure
K. Matoba: Research Support; Self; Sanofi K.K., Takeda Pharmaceutical Company Limited. Y. Takeda: None. Y. Nagai: None. T. Akamine: None. Y. Kanazawa: None. D. Kawanami: None. K. Utsunomiya: None. R. Nishimura: Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc. Speaker’s Bureau; Self; Abbott, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly Japan K.K., Medtronic, Merck & Co., Inc., Novo Nordisk Inc., Sanofi K.K., Takeda Pharmaceutical Company Limited.

Funding
Japan Society for the Promotion of Science (18K15985)
492-P: Associations between Novel Biomarkers of Inflammation and Estimated Glomerular Filtration Rate in Adults with Recent-Onset Type 1 and Type 2 Diabetes
Authors: MAALMI HAIFA , HERDER CHRISTIAN , STRASSBURGER KLAUS , URNER SOFIA , JANDELEIT-DAHM KARIN , ZAHARIA OANA P. , KARUSHEVA YANISLAVA , BODIS KALMAN , RATHMANN WOLFGANG , MARKGRAF DANIEL F. , BURKART VOLKER , SZENDROEDI JULIA , RODEN MICHAEL 
Published: June 2020

Background: The development of anti-inflammatory therapies for renal function loss in patients with diabetes requires identification of specific biomarkers which might differ by diabetes type. This study aimed to investigate associations between a novel panel of biomarkers of inflammation and estimated glomerular filtration rate (eGFR) in patients with type 1 diabetes (T1D) and type 2 diabetes (T2D).
Methods: This study was based on 145 participants with T1D (mean age 33.8 years, 60.7% males) and 250 participants with T2D (mean age 55.3 years, 66.4% males) from the observational prospective German Diabetes Study (GDS), which includes adults with a diabetes duration &lt;1 year. A panel of 74 biomarkers of inflammation was measured in serum of fasted participants using the proximity extension assay technology. eGFR was calculated from serum creatinine and cystatin C using the CKD-EPI equation. Associations between each biomarker and eGFR were assessed using multiple linear regression adjusting for age, sex, BMI, HbA1c, diabetes duration, lipids, hypertension and use of NSAIDs and glucose-lowering drugs.
Results: Baseline eGFR was higher in T1D compared to T2D (mean (SD) 101.8 (15.3) vs. 89.3 (15.6) mL/min/1.73 m2; P&lt;0.0001). After correction for multiple testing, 6 and 24 biomarkers were inversely associated with eGFR in T1D and T2D, respectively (all P&lt;0.05). TNFB was T1D-specific, while ADA, CD40, CD244, CCL3, CCL20, CCL25, CX3CL1, CXCL9, CSF1, CST5, FGF23, FGF5, IL-15RA, IL12B, MMP10, NT3, SCF, TGFα and uPA were T2D-specific. CCL23, CD5, IL10RB, PDL1 and TNFRSF9 were associated with eGFR in both diabetes types.
Conclusion: We identified novel inflammatory biomarkers, independently associated with eGFR in adults with recently diagnosed T1D and T2D. As a next step, these biomarkers shall be evaluated as potential predictors of incident and progressive diabetic nephropathy.

Disclosure
H. Maalmi: None. C. Herder: Research Support; Self; Sanofi-Aventis. K. Strassburger: None. S. Urner: None. K. Jandeleit-Dahm: None. O.P. Zaharia: None. Y. Karusheva: None. K. Bodis: None. W. Rathmann: Advisory Panel; Self; AstraZeneca. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc. Research Support; Self; Novo Nordisk Inc. Speaker’s Bureau; Self; Eli Lilly and Company, Novo Nordisk Inc. D.F. Markgraf: None. V. Burkart: None. J. Szendroedi: None. M. Roden: Advisory Panel; Self; Servier. Board Member; Self; Poxel SA. Consultant; Self; Eli Lilly and Company, Gilead Sciences, Inc., ProSciento, TARGET PharmaSolutions. Research Support; Self; Boehringer Ingelheim International GmbH, Novartis Pharma K.K., Sanofi US. Speaker’s Bureau; Self; Novo Nordisk A/S.
493-P: Cardiac Microvascular Endothelial Enhancement of Cardiomyocyte Function Is Impaired by Uremic Serum and Restored by Empagliflozin
Authors: JUNI RIO , AL-SHAMA RUSHD , KUSTER DIEDERIK , DER VELDEN JOLANDA VAN , VERVLOET MARC , PAULUS WALTER , ERINGA ETTO C. , KOOLWIJK PIETER , VAN HINSBERGH VICTOR W. 
Published: June 2020

Background: It has been proposed that diabetes and chronic kidney disease cause heart failure with preserved ejection fraction (HFpEF) through low-grade inflammation and microvascular endothelial dysfunction. We have previously shown that endothelial enhancement of cardiomyocyte contraction and relaxation is mediated by NO, impaired by cytokines, and restored upon treatment with the SGLT2 inhibitor Empagliflozin. Chronic kidney disease (CKD) is linked to diabetes and HFpEF and causes systemic endothelial dysfunction.
Objective: To test the hypothesis that uremic serum from patients with CKD impairs endothelial control of cardiomyocyte contraction and relaxation, and that Empagliflozin protects against this effect.
Methods and Results: Co-culture of rat cardiomyocytes (CM) with human cardiac microvascular endothelial cells (CMECs) enhanced their contraction and relaxation. Pre-treatment with serum from patients with CKD abrogated these effects of CMEC, and treatment of CMECs with Empagliflozin restored the effects of CMECs on CM function. Exposure of CMECs to the uremic serum reduced endothelial NO bioavailability and increased intracellular reactive oxygen species ROS by increasing ROS generation by mitochondria. Uremic serum also increased 3-nitrotyrosine (3-NT) level, indicating scavenging of NO by ROS. Empagliflozin attenuated uremic serum-induced mitochondrial and intracellular ROS, leading to restoration of endothelial NO bioavailability.
Conclusions: Exposure of CMECs to serum from CKD patients impairs CMEC-mediated enhancement of CM contraction and relaxation. Empagliflozin restores the beneficial effect of CMECs on CM function by reducing mitochondrial oxidative damage, leading to a decrease in intracellular ROS accumulation and increased endothelial NO bioavailability.

Disclosure
R. Juni: None. R. Al-Shama: None. D. Kuster: None. J. van der Velden: None. M. Vervloet: None. W. Paulus: None. E.C. Eringa: None. P. Koolwijk: None. V.W. van Hinsbergh: None.

Funding
Netherlands Heart Foundation
5-LB: Consuming High-Fructose Corn Syrup or Sucrose-Sweetened Beverages Increases Hepatic Lipid Content and Decreases Insulin Sensitivity in Young Adults
Authors: SIGALA DESIREE , HIERONIMUS BETTINA , PRICE CANDICE , LEE VIVIEN , NUNEZ MARINELLE , MEDICI VALENTINA , CHAUDHARI ABHIJIT J. , MCGAHAN JOHN P. , ABDELHAFEZ YASSER , KEIM NANCY L. , GORAN MICHAEL I. , PACINI GIOVANNI , TURA ANDREA , HAVEL PETER J. , STANHOPE KIMBER L. 
Published: June 2020

Rodent studies demonstrate that compared with diets sweetened with sucrose, high fructose corn syrup (HFCS) produces greater metabolic dysfunction. However, the effects of sustained consumption of HFCS and sucrose have not been adequately compared in humans. The objective of this study was to compare HFCS-, sucrose-, or aspartame- sweetened beverage (-SB) consumption on hepatic lipid content and insulin sensitivity in young adults. In a parallel, double-blinded study, adult men and women (18-40 yrs) were assigned to beverage groups matched for sex, BMI and fasting triglycerides. Inpatient magnetic resonance imaging for hepatic lipid content and oral glucose tolerance tests were performed at baseline and intervention 15 days later. For 12 outpatient days subjects consumed their usual diets with 3 servings/day of HFCS- (n=28) or sucrose-SB (n=24) providing 25% energy requirement (Ereq), or aspartame-SB (n=23). Matched standardized diets were provided during inpatient testing with the sugar in beverages replacing complex carbohydrate during intervention.
Hepatic lipid content was increased by consumption of HFCS-SB (LS means of absolute Δ±SEM: 0.5%±0.2, P&lt;0.05) and sucrose-SB (Δ: 0.5%±0.2, P&lt;0.01) compared with baseline levels and compared with aspartame-SB (Δ: -0.2%±0.2, P&lt;0.05 vs. HFCS- or sucrose-SB). The 3-hr Matsuda insulin sensitivity index was decreased by consumption of HFCS-SB (Δ: -0.3±0.2) and sucrose-SB (Δ: -0.5±0.2, P&lt;0.01 vs. baseline) compared with aspartame-SB (Δ: 0.3±0.2, P&lt;0.05 vs. HFCS- or sucrose-SB). The 2-hr Oral Glucose Insulin Sensitivity index was also decreased by consumption of HFCS-SB (Δ: -13.6±7.5) and sucrose-SB (Δ:-11.5±7.9) compared with aspartame-SB (Δ: +17.9±8.3, P&lt;0.05 vs. HFCS- or sucrose-SB).
Consumption of SB providing 25% of Ereq as HFCS or sucrose for 2 weeks increased hepatic lipid content and reduced insulin sensitivity in young adults compared with consumption of aspartame-SB.

Disclosure
D. Sigala: None. B. Hieronimus: None. C. Price: None. V. Lee: None. M. Nunez: None. V. Medici: None. A.J. Chaudhari: None. J.P. McGahan: None. Y. Abdelhafez: None. N.L. Keim: None. M.I. Goran: Advisory Panel; Self; Yumi. Other Relationship; Self; Penguin Random House. G. Pacini: None. A. Tura: None. P.J. Havel: Research Support; Self; Arrowhead Pharmaceuticals, Inc., Magnamosis, Inc. Other Relationship; Self; None. K.L. Stanhope: None.

Funding
National Institutes of Health (R01HL091333, R01HL107256)
5-OR: Prior Bariatric Surgery and Perioperative Cardiovascular Outcomes following Noncardiac Surgery in Patients with Type 2 Diabetes Mellitus
Authors: JIN JIEWEN , LI HAI , ZHANG PENGYUAN , LI YANBING 
Published: June 2020

Background: Diabetes mellitus is a risk factor for perioperative major adverse events. This study aims to evaluate the association between prior bariatric surgery (prior-BS) and perioperative cardiovascular outcomes following noncardiac surgery in patients with type 2 diabetes mellitus (T2DM).
Methods: We used the National Inpatient Database to identify major noncardiac surgery in T2DM patients from 2006 to 2014. The primary outcome is major perioperative adverse cardiovascular and cerebrovascular events (MACCE), which included death, acute myocardial infarction and acute ischemic stroke. In-hospital outcomes between patients with prior-BS and morbid obesity were compared using unadjusted logistic, multivariable logistic and propensity score matching analyses.
Results: A weighted of 152,6820 patients diagnosed with T2DM who underwent noncardiac surgery were included. Both rates of prior-BS and morbid obesity significantly increased during the study period (P&lt;0.0001). Patients with prior-BS were younger, more likely to be female and had more smoking, alcohol abuse, anemia and prior venous thromboembolism and prior percutaneous coronary intervention. Incidence of MACCE was 1.01% and 3.25% in patients with prior-BS and morbid obesity, respectively. After multivariable adjustment, we found that prior-BS was associated with reduced risk of MACCE (odds ratio[OR]: 0.71; 95% confidence interval [CL]: 0.62-0.81), death (OR: 0.64; 95%CI: 0.52-0.78), acute kidney injury (OR: 0.66; 95%CI: 0.62-0.70) and acute respiratory failure (OR: 0.46; 95%CI: 0.42-0.50).
Conclusions: Prior bariatric surgery in diabetes mellitus patients who underwent noncardiac surgery is associated with lower risk of MACCE. Prospective studies are need to verify the benefits of bariatric surgery in noncardiac surgery.

Disclosure
J. Jin: None. H. Li: None. P. Zhang: None. Y. Li: None.

Funding
China Postdoctoral Science Foundation (2018M640871)