bluuebunny/crossref_metadata_2025 huggingface dataset:
444-P: Evaluating Factors Associated with Cardiovascular and Cerebrovascular Complications of Type 2 Diabetes
Authors: PREWITT TODD G. , MANNER DAVID , DECLUE RICHARD W. , NAIR RADHIKA , BAJPAI SANJAY K. , WONG-JACOBSON SIEW HOONG , JUNEJA RATTAN , PENG XIAOMEI
Published: June 2019

Our aim was to understand characteristics of patients with type 2 diabetes mellitus (T2DM) who develop macrovascular complications. This was a retrospective observational study using administrative claims from a large national health plan. We identified patients with T2DM between January 2010 and June 2012; date of the first claim for T2DM was set as the index date. Patients 18-89 years of age enrolled in a health plan for at least one year prior (baseline) and post-index, with at least one HbA1c value during pre-index, were identified. Patients were categorized into incident and prevalent cohorts of T2DM based on absence/presence of claims during the baseline period. Patients with cerebrovascular and cardiovascular complications (CV complications), as defined by the diabetes complications severity index, were evaluated. Descriptive analyses were used to evaluate characteristics of patients with and without CV complications. Landmark dynamic risk modeling was used to identify factors predicting CV complications for those with no history of CV complications during the baseline period. We identified 111,205 patients with T2DM; 61,101 (54.6%) had no history of CV complications at baseline. During the five years post-index, 13,059 (21.4%) and 31,472 (51.5%) had a cerebrovascular or cardiovascular complication, respectively. Proportions were slightly higher in the prevalent T2DM cohort, especially for cerebrovascular complications (22.3% vs. 20.2%, P<0.001). For patients with no history of CV complications, landmark dynamic risk modeling identified smoking (HR 1.26), obesity/overweight (HR 1.14), hypertension (HR 1.27), dyslipidemia (HR 1.11) and peripheral vascular disease (HR 1.19) increased risk of CV complications (P<0.001). Identifying modifiable variables that change over time, and the presence of chronic comorbidities, can aide development of specific interventions to evaluate and treat patients with T2DM, while addressing risk of CV complications.

Disclosure

T.G. Prewitt: None. D. Manner: Employee; Self; Eli Lilly and Company. R.W. DeClue: Employee; Self; Humana. R. Nair: Stock/Shareholder; Self; Abbott Laboratories, AbbVie Inc., GlaxoSmithKline plc. S.K. Bajpai: Employee; Self; Eli Lilly and Company. S. Wong-Jacobson: None. R. Juneja: Employee; Self; Eli Lilly and Company. X. Peng: None.

Funding

Eli Lilly and Company
452-P: Albumin/Creatinine Ratio and Pulse Wave Velocity Predicts Coronary Artery Disease and Renal Outcome in Type 2 Diabetic and Prediabetic Subjects
Authors: SANCHEZ RAMIRO A. , SANCHEZ MARIA J. , RAMIREZ AGUSTIN J.
Published: June 2019

Introduction: Silent coronary heart disease is frequently undetected in type 2 diabetes mellitus (DM2) and prediabetes determined by glucose intolerance (GI). Pulse wave velocity (PWV) and albumin-creatinine ratio (ACR) have been considered markers of cardiovascular mortality, coronary heart disease and chronic renal failure.

Aims: To evaluate the incidence of coronary heart disease (CHD) and its relationship with urinary albumin-creatinine ratio, glomerular filtration rate (GFR) and PWV in type 2 DM with silent CHD.

Methods: We analyzed 92 individuals (44 male), 49 (60±7y) type 2 DM non-insulin dependents and 43 prediabetics (43±4y), with Grade I-II hypertension and no symptoms of CHD. All type 2 DM patients were under hypoglycemic treatment with an A1C hemoglobin between 5.5 and 6.5%. Every patient underwent a myocardial perfusion SPECT scan. In those subjects with ischemic patterns, a coronary angiography was performed. In addition, a PWV, glomerular filtration rate (CrCl), and ACR were evaluated. Statistics: mean±SEM, and ANOVA followed by Post Hoc analysis among groups.

Results: Silent CHD was found in 48.59% of DM2 and 25.58% of GI. DM2 and GI patients with CHD had higher ACR and PWV and reduced glomerular filtration rate (p<0.05). DM2 and GI showed a negative relationship between GFR and ACR (p<0.05). Type 2 diabetic and GI showed a negative relation between GFR and ACR or PWV when having GFR above 60 mil.min. Meanwhile, in type 2 Diabetics with CHD and GFR < 60 ml.min a negative relationship with PWV was only, suggesting a cardiorenal interaction in type 2 diabetes.

Conclusions: Higher PWV, lower GFR and ACR predict the incidence of CHD in DM2. GI also represent a group of higher risk for coronary heart disease with the same predictors showed in DM2. PWV seems to represent a reliable marker.

Disclosure

R.A. Sanchez: None. M.J. Sanchez: None. A.J. Ramirez: None.
46-OR: A Novel Interaction between AT1 Receptor and Cytochrome-p450 Lipid Metabolites in Diabetic Retinopathy
Authors: AL-SHABRAWEY MOHAMED , IBRAHIM AHMED , TAWFIK AMANY M. , HENG WANG MONG HENG
Published: June 2019

Nearly all patients with type 1 diabetes and 60% of those with type 2 diabetes develop diabetic retinopathy (DR). Thus, there is an urgent need to identify new therapeutic targets to prevent or at least to minimize microvascular dysfunction associated with DR. Activation of the renin-angiotensin system (RAS) contributes to breakdown of blood-retinal barrier (BRB) and neovascularization in DR. Clinical studies underscored RAS as a promising therapeutic target to treat DR. Although, blockade of RAS has reduced the incidence of DR in clinical studies, it did not reduce the progression of DR. Cytochrome-P450 metabolizes arachidonic acid to the pro-angiogenic and -permeability mediators, epoxyeicosatrienoic acids (EETs) which are metabolized by the soluble epoxide hydrolase (sEH) to inactive products. Our experiments demonstrated that:

1.) Angiotensin II (Ang II) increases retinal expression of sEH by 2.8 fold (2.84 ± 1.1 vs. 1 ± 0.08, P < 0.05, n = 4), which is blunted by angiotensin receptor-1 (AT1) inhibition;

2.) both sEH blockade and knockout (KO), which accumulate EETs, enhance Ang II-induced BRB injury as shown by 1.58 fold increase in retinal albumin leakage (1.58 ± 0.4 vs. 1 ± 0.13, P < 0.05, n = 4) and 2.8 fold (2.8 ± 1.7 vs. 1 ± 0.09, P < 0.05, n = 4), respectively. These results were confirmed by fluorescein angiogram analysis;

3.) Administration of synthetic 11,12-EET enhances intravitreal Ang II-induced retinal albumin leakage by 2.2 fold (2.2 ± 0.5 vs. 1 ± 0.18, P < 0.05, n = 4); and

4.) sEH KO potentiates diabetes (STZ)-induced retinal damage. This was associated with upregulation of retinal vascular endothelial growth factor (VEGF) and glucose transporter-1 (GLUT-1).

In conclusion, our data demonstrate that in DR, activation of retinal AT1 triggers a compensatory mechanism by increasing retinal sEH that decreases EETs production. Thus, the use of combined EETs blockade with AT1 blocker holds the promise, as a new therapeutic strategy to prevent or reduce DR.

Disclosure

M. Al-Shabrawey: None. A. Ibrahim: None. A.M. Tawfik: None. M.H. Wang: None.

Funding

American Heart Association (SE00144); National Institutes of Health (R01EY023315 to M.A-S.)
462-P: Diabetes Is Associated with Progression of Coronary Arterial Calcification in Patients New to Dialysis
Authors: ROY NEIL , YANG DANWEN , ROSAS SYLVIA
Published: June 2019

Background and Objective: Coronary artery calcification is a risk marker of atherosclerosis. Recently, a large multicenter, prospective observational study showed the CAC density was inversely related to the occurrence of cardiac vascular events. The proposed hypothesis is that low density area is more prone to rupture due to large lipid core and a marker of plaque instability. However, high CAC density in 125 chronic hemodialysis patients was associated with increased mortality. Our aim was to determine CAC density in new dialysis patients, and the role of DM in the progression of CAC density.

Design, Setting, Participants, and Measurements: Ninety-eight initial dialysis patients were enrolled. CAC density was calculated by Agatson score divided by calcium volume score. CAC density progression was measured by the difference in score between CT/time. Sixty-seven study participants had repeat CAC measures at one year.

Results: The mean age of study participants was 50.1 ± 12.7 years. A third were women, and 64.7% were black. The baseline CAC density for DM group was 85.2 (interquartile range (IQR), 10.3-452) compared to 0(IQR, 0-90.3) for non-DM group was (P=0.01). Seventy percent of diabetes patients had progression of calcium density while forty one percent of nondiabetic patients had progression of calcium density score. (P=0.02). Hazard ratio for DM after adjust for age and smoke is 2.0 (1.05-72.6).

Conclusions: Our results indicate that DM is a risk factor for high calcium density of individuals new to dialysis patients and is a risk factor for progression of calcium density score. Increased calcium density in new dialysis patient is a risk factor for CAC progression, rather than a marker of plaque stabilization.

Disclosure

N. Roy: None. D. Yang: None. S. Rosas: Research Support; Self; Bayer US, Ironwood Pharmaceuticals, Inc., Janssen Pharmaceuticals, Inc.
468-P: Nonalcoholic Steatohepatitis Is Associated with Diabetic Cardiomyopathy in Type 2 Diabetes
Authors: CHO DONG-HYEOK
Published: June 2019

Background: Abnormalities in cardiac structure and function in type 2 diabetic patients may develop in the absence of ischemic heart disease. These abnormalities are attributed to diabetic cardiomyopathy. Impaired diastolic heart function has been observed in persons with nonalcoholic fatty liver disease (NAFLD) and/or with type 2 diabetes. We investigated the association between liver fibrosis and left ventricular (LV) diastolic dysfunction in type 2 diabetes.

Methods: We studied 92 patients with type 2 diabetes (51 men; mean age 62 ± 6 years) who had undergone liver ultrasonography and conventional Doppler echocardiography. Presence of NAFLD and/or advanced liver fibrosis was determined by abdominal ultrasonography and NAFLD fibrosis score (NFS). LV diastolic dysfunction was defined according to transmitral peak early to late ventricular filling (E/A) ratio and deceleration time (DT), using echocardiography.

Results: Fifty-four patients (58.7%) had NAFLD. On echocardiography, LVEF was within normal range in both groups, whereas LV mass index (P=0.035) and LA diameter (P=0.021) were significantly greater in the NAFLD patients. The systemic vascular resistance and arterial elasticity were not different. NAFLD patients had lower E/A ratio (P=0.014) and longer DT (P=0.042) than those without steatosis. When NAFLD was stratified by NFS, subjects with advanced liver fibrosis exhibited a higher prevalence of diastolic dysfunction (46.2%, 55.6%, 60.0%; none, simple steatosis, advanced fibrosis, respectively; P for trend = 0.025). In multivariate logistic regression, liver fibrosis was independently associated with diastolic dysfunction (OR=1.46, 95% CI=1.02-2.46, P=0.041) after adjusting for cardiometabolic risk factors.

Conclusions: Our data show that in patients with type 2 diabetes and NAFLD, liver fibrosis was associated with LV diastolic dysfunction and may be an independent risk factor for diastolic dysfunction.

Disclosure

D. Cho: None.
476-P: Pemafibrate, a PPAR Alpha Agonist, Attenuates Neointima Formation after Vascular Injury in Nondiabetic and Diabetic Mice
Authors: HORIKAWA TSUYOSHI , NOMIYAMA TAKASHI , TAKAHASHI HIROYUKI , KAWANAMI TAKAKO , HAMAGUCHI YURIKO , TANAKA TOMOKO , YANASE TOSHIHIKO
Published: June 2019

Recently, the prevention of cardiovascular events has become one of most important purpose of diabetes care. We have previously reported that GLP-1 receptor agonist exendin-4 (BBRC 2011) and DPP-4 inhibitor linagliptin (Cardiovasc Diabetol 2014) attenuate neiontima formation after vascular injury. On the other hand, PPAR agonist is also reported its vascular protective effect. Then, we examined whether selective PPAR alpha agonist pemafibrate could attenuate neointima formation after vascular injury and vascular smooth muscle cell (VSMC) proliferation. We performed endothelial denudation injury in mice treated with high fat diet (HFD) or normal chow. Orally administered pemafibrate significantly attenuated neointima formation after vascular injury in both HFD and normal chow mice. Interestingly, pemafibrate increased serum FGF-21 concentration and decreased serum insulin concentration in HFD mice. In addition, body weight was slightly but significantly decreased by pemafibrate in HFD mice. In vitro experiments, pemafibrate attenuated VSMC proliferation in growth curve in a dose dependent manner, but not another PPAR alpha agonist bezafibrate. This suggested that the attenuation of VSMC proliferation by pemafibrate could be the drug effect, not class effect of PPAR alpha agonist. BrdU assay revealed that pemafibrate inhibited DNA synthesis in VSMC dose-dependently. In cell cycle distribution, G1 to S phase entry was significantly inhibited by pemafibrate in flow cytometry analysis. However, apoptosis was not induced by pemafibrate in VSMC in TUNEL assay. These data suggested that pemafibrate attenuates neiontima formation after vascular injury and VSMC proliferation inhibiting cell cycle progression.

Disclosure

T. Horikawa: None. T. Nomiyama: None. H. Takahashi: None. T. Kawanami: None. Y. Hamaguchi: None. T. Tanaka: None. T. Yanase: None.
480-P: Diabetes and Experimental Myocardial Infarction Synergize to Accelerate Atherosclerosis Destabilization
Authors: KRAMER FARAH , MARTINSON AMY M. , PAPAYANNOPOULOU THALIA , MURRY CHARLES E. , KANTER JENNY E.
Published: June 2019

In addition to increasing the risk of an initial myocardial infarction (MI), diabetes increases the risk of a recurrent MI. Work by others suggests that an experimental MI can accelerate atherosclerosis via monocytosis. To test if diabetes and experimental myocardial infarction synergize to accelerate atherosclerosis, we induced atherosclerosis in Ldlr-/-; Gp+ mice, lethally irradiated them and transplanted with either wild type (WT) or LysM-Cre-mediated CD49D-deficient bone marrow. Following recovery, diabetes was induced using lymphocytic choriomeningitis virus, or saline was used as a control. The mice were subjected to ligation of the left anterior descending coronary artery to induce experimental MI or a sham surgery and then maintained for an additional 3 weeks. Diabetic (D) mice exhibited hyperglycemia and elevated plasma cholesterol and triglyceride levels compared to nondiabetic (ND) mice, but neither hematopoietic genotype or MI influenced that. All mice subjected to MI had significantly reduced left ventricular function. Neither diabetes nor MI resulted in monocytosis compared to ND sham mice. Despite not affecting total lesion size, the combination of diabetes and MI resulted in larger necrotic cores in the aortic sinus and in the brachiocephalic artery (9.8 ± 1% of total lesion in aortic sinus in ND sham, 11.9 ± 1% in ND MI, 15.4 ± 1% in D sham and 18.1 ± 2% in D MI, P<0.001 comparing ND Sham to D MI, n=14-24, ANOVA). The expansion of the necrotic cores was dependent on monocyte recruitment, since mice with myeloid cells deficient in the adhesion molecule CD49D were protected from necrotic core expansion. Diabetes and MI also synergized to accelerate macrophage cell death (6.1 ± 3% of peritoneal macrophages were dead in ND sham, 6.1 ± 3% in ND MI, 11± 1% in D sham and 20 ± 3% in D MI, n=3-6, p<0.05 ANOVA).

In summary, diabetes and experimental MI destabilize the atherosclerotic lesion, potentially explaining why patients with diabetes are at an increased risk of recurrent MI.

Disclosure

F. Kramer: None. A.M. Martinson: None. T. Papayannopoulou: None. C.E. Murry: Other Relationship; Self; Cytocardia. J.E. Kanter: None.

Funding

American Diabetes Association (1-16-IBS-153 to J.E.K.)
481-P: KCNMA1 Transcriptional Regulation by Nrf2 in Diabetic Vessels
Authors: LEE HON-CHI , SUN XIAOJING , WANG XIAO-LI , LU TONG
Published: June 2019

KCNMA1 encodes the large conductance Ca2+-activated K+ channel pore-forming subunits (BK-α), a major ionic mechanism that regulates vascular smooth muscle physiology and is impaired in high fat diet-induced (HFD) diabetic mice. The nuclear factor erythroid 2-related factor 2 (Nrf2) is a master transcription factor that regulates a wide variety of cellular proteins by binding to the antioxidant response elements (AREs). We found that the promoter region of KCNMA1 contains multiple AREs, but whether Nrf2 regulates KCNMA1 expression is unknown. In this study, we hypothesize that Nrf2 is a transcriptional regulator of KCNMA1 and the reduced Nrf2 activity in diabetes contributes to BK channelopathy. Using videomicroscopy, patch clamp recordings, molecular and cellular approaches, we found that BK channel activity and BK channel-mediated vasodilation were diminished in the coronary arteries of mice after 4 months on HFD. This was accompanied by significant reductions in BK-α and Nrf2 protein expression. In cultured coronary artery smooth muscle cells (CASMCs), adenoviral transduction with Nrf2 (50 MOI) resulted in an increase of BK-α mRNA by 10-fold and BK-α protein expression by 2-fold, while a 70% knockdown of Nrf2 by shRNA produced a 30% downregulation of BK-α protein levels. Luciferase reporter assays showed robust luciferase signals in cells 48 h after co-transfection with Nrf2 vectors and reporter vectors containing a segment of the KCNMA1 promoter region with a predicted ARE-Nrf2 binding site, but not those with empty vectors or KCNMA1 reporter vectors with the ARE-Nrf2 binding site deleted. Most importantly, treatment with dimethyl fumarate (a Nrf2 activator) increased BK-α mRNA and protein expression in cultured CASMCs, and rescued BK channel function and BK channel-mediated vasodilation in coronary arteries of HFD mice. Our results indicate that Nrf2 transcriptionally regulates KCNMA1 expression in CASMCs and has a therapeutic potential in the treatment of BK channelopathy and vasculopathy in diabetes.

Disclosure

H. Lee: None. X. Sun: None. X. Wang: None. T. Lu: None.

Funding

American Diabetes Association (1-18-IBS-210 to H-C.L.); National Institutes of Health (R01HL74180)
490-P: Administration of AAV-Containing Glyoxalase-I Attenuated Nephropathy in Rats with Chronic Diabetes
Authors: BIDASEE KESHORE , ALOMAR FADHEL AHMED
Published: June 2019

Supra-physiologic flux/production of methylglyoxal (MG) has emerged as an underlying cause of nephropathy in chronic type 1 diabetes mellitus (T1DM). This increased MG arises in part from a reduction in expression of its detoxifying enzyme, glyoxalase-I (Glo-I). Adeno-associated virus (AAV)-mediated gene transfer has materialized as a promising strategy to increase expression of deficit proteins in disease states. Here we compared the efficacies of AAV2/9 containing Glo-I driven by two promoters, endothelin-1 (AAV2/9-Endo-Glo-I) and cytomegalovirus (AAV2/9-CMV-GloI) to attenuate nephropathy in T1DM rat. After eight weeks of T1DM, urine production increased 4-fold, urine norepinephrine increased 2.5-fold, serum creatinine increased 4-fold, creatinine clearance decreased 4-fold, BUN increased 5-fold, and proteinuria increased 8-fold. Renal Glo-I protein was 50% lower, MG was 2.5-fold higher, and extravasation of albumin from peritubular and glomerular capillaries and fibrosis were prominent. TNF-α and NF-κB (p65) phosphorylation were increased 3-4 folds. A single bolus injection of AAV/2/9 Glo-I and AAV2/9-Glo-I (1.7×1012 viron particles/kg) ten days after T1DM onset blunted increases in water consumption and urine assayed eight weeks later. AAV2/9-Glo-I also attenuated increases in norepinephrine, creatinine, BUN, proteinuria, albumin extravasation from renal microvessels, fibrosis, MG, TNF-α, NF-κB, activation and increased creatinine clearance rate. The magnitudes of the effects were higher in AAV2/9-Endo-Glo-I injected rats in part to higher expression of Glo-I in tubules and reduced expression of the MG-synthesizing enzyme vascular adhesion protein-1 within glomeruli. These new data demonstrate that intravenous administration of AAV2/9-containing Glo-I after the onset of T1DM attenuated renal dysfunction in rats via multiple mechanisms involving reductions in sympatho-excitation, inflammation, microvascular leakage and fibrosis.

Disclosure

K. Bidasee: None. F.A. Alomar: None.

Funding

National Institutes of Health (P30GM103335); University of Nebraska Medical Center (GTRP 1053); UNeMed
494-P: Whole-Kidney 3D Imaging and Image Analysis Enables Detailed Quantification of Diabetic Nephropathy Features in the Uninephrectomized db/db Mouse Model
Authors: FINK LISBETH N. , SEMBACH FREDERIKKE E. , ROOSTALU URMAS , JOHANSEN THEA , SKYTTE JACOB LERCKE , SECHER THOMAS , FOSGERAU KELD , VRANG NIELS , JELSING JACOB , HECKSHER-SØRENSEN JACOB , PEDERSEN TANJA X.
Published: June 2019

Diabetic nephropathy (DN) is a serious complication of diabetes and the leading cause of end-stage renal disease. To reproducibly test novel therapies, there is a need for precise quantification of DN pathology in rodent models. We compared the accelerated development of DN in male and female uninephrectomized db/db mice using advanced imaging techniques. Unilateral nephrectomy (UNx) was performed in 7-8 weeks old male and female db/db mice. Sham-operated db/+ mice served as controls. Kidneys were preserved for histology and stereology. In a separate study, UNx was performed in 18 weeks old male db/db mice. These mice and control db/+ mice were injected with lectin-594 prior to termination to visualize glomerular morphology by 3D light sheet microscopy. All mice were terminated at 24 weeks of age. Male and female db/db UNx mice showed comparable increases in albuminuria. Total glomerular volume was markedly higher in male and female db/db UNx mice compared to db/+ controls (mean males; 4.42 vs. 2.71 mm3; p<0.001; females; 5.04 vs. 2.69 mm3; p<0.001). Tubulointerstitial collagen III immuno-staining was significantly increased only in female db/db UNx mice compared to db/+ mice. In contrast, total glomerular collagen IV levels were increased in male db/db UNx mice compared to db/+ controls (mean 1.30 vs. 0.80 mm3; p<0.01), and female db/db UNx mice displayed increased glomerular collagen IV compared to male db/db UNx mice (mean 1.80 vs. 1.30 mm3; p<0.05). 3D imaging at 24 weeks following UNx at 18 weeks of age confirmed expansion in glomerular volume (median db/+ 96.000 vs. db/db 145.000 µm3). Moreover, a marked increase was seen in cortical lectin-594 staining, indicating glomerular leakage in db/db UNx mice.

In conclusion, male and female db/db UNx mice displayed comparable features of DN and both constitute valid models for DN. The use of whole-kidney 3D imaging provides a novel avenue for assessment of individual glomerular volume and permeability.

Disclosure

L.N. Fink: Employee; Self; Gubra. Stock/Shareholder; Self; Novo Nordisk A/S. F.E. Sembach: Consultant; Self; Novo Nordisk A/S. Employee; Self; Gubra. U. Roostalu: Employee; Self; Gubra. T. Johansen: Employee; Self; Gubra. J.L. Skytte: Employee; Self; Gubra. T. Secher: Employee; Self; Gubra. Employee; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Self; Gubra. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. K. Fosgerau: Stock/Shareholder; Self; Gubra. N. Vrang: Board Member; Self; Gubra. Stock/Shareholder; Self; Gubra. J. Jelsing: Stock/Shareholder; Self; Gubra. J. Hecksher-Sørensen: Employee; Self; Gubra. Stock/Shareholder; Self; Novo Nordisk A/S. T.X. Pedersen: Employee; Self; Gubra.
497-P: Fyn-Dependent Tyrosine Phosphorylation of Tgm2 Modulates p53-Tgm2-p62 Complex in Renal Proximal Tubular Epithelial Cells
Authors: YAMADA EIJIRO , OKADA SHUICHI , BASTIE CLAIRE C. , YAMADA MASANOBU
Published: June 2019

Diabetic kidney disease (DKD) is one of the major diabetic complications, the leading cause of the end stage renal disease (ESRD). Recently autophagy, which is also implicated in diabetes and insulin resistance, was shown to regulate DKD. Previously, we reported that Fyn functions as a muscle mass regulator by suppressing autophagy but it is still not known whether Fyn regulates autophagy in kidney and functions as a pathogenesis of DKD. Recently, we demonstrated that over-expressing of Fyn in HK2 cells as an in vitro cell model of renal proximal tubular epithelial cells inhibits autophagy. SiRNA mediating knockdown of Fyn increased autophagy demonstrating Fyn has important role for autophagy in HK2 cells. Phospho-proteomic analysis revealed that Fyn phosphorylates Transglutaminase 2 (Tgm2), a known autophagic inhibitor through Beclin1, on Y369 and Y617. More recently, we demonstrated that Fyn dependent phosphorylation of Tgm2 could regulate autophagy in HK2 cells.

While it has been reported that Tgm2 could cross-link Beclin1 to inhibits its activity, no cross-linking of Beclin1 was seen in Tgm2 expressing HK2 cells, indicating there could be another mechanism of Fyn-Tgm2 regulating autophagy in HK2 cells. Recently, Tgm2 was shown to make a complex with p53 and p62, known autophagy regulator/substrate to degrade p53 in autophagosome in cancer cells. Interestingly, we found that p53 expression was decreased in Tgm2 knock-downed HK2 cells and increased in Tgm2 expressing HK2 cells. More importantly co-immunoprecipitation assay showed that Tgm2-Y369,617F which could not be phosphorylated by Fyn bind to p53 more than WT and dissociated from p62. Taken together, these data suggest that Fyn phosphorylates Tgm2 on Y369, 617 to modulate p53-Tgm2-p62 complex in the renal tubulus cells leading to a pathogenesis of DKD.

Disclosure

E. Yamada: None. S. Okada: None. C.C. Bastie: None. M. Yamada: None.
50-OR: Identifying Risk of Blindness in Patients with Diabetic Retinopathy
Authors: NGUYEN QUAN DONG , HALL REBECCA , KELLY SCOTT P. , LUM FLORA , STOILOV IVAYLO , ABBASS IBRAHIM M. , TO TU MY , ABOLIAN ANNA , GARMO VINCENT
Published: June 2019

Purpose: Using real-world data from the American Academy of Ophthalmology (Academy) IRIS® Registry (Intelligent Research in Sight), this retrospective study sought to identify risk factors for developing sustained blindness (SB) in eyes with newly diagnosed diabetic retinopathy (DR).

Methods: IRIS Registry data were reviewed to identify eyes newly diagnosed with DR between Jan 1, 2013 and Dec 31, 2017 (index date). Included eyes had ≥ 2 visual acuity (VA) readings of 20/40 or better before or ≤ 3 months post index; eyes with evidence of prior DR, retinal disease or retinal treatment were excluded. Patients were followed until the development of SB (defined as ≥ 2 VA readings of 20/200 or worse ≥ 3 months apart), or most recent VA reading. Kaplan-Meier analyses estimated time to SB by DR severity; multivariable Cox proportional hazards models assessed the impact of baseline characteristics and ocular conditions during follow-up on the risk of developing SB.

Results: A total of 53,535 eyes were analyzed, 678 of which developed SB during follow-up. At index, 49.4% of eyes were diagnosed with mild non-proliferative DR (NPDR), 9.1% moderate NPDR, 1.5% severe NPDR, 10.5% proliferative DR (PDR), and 29.5% unspecified DR. Relative to eyes with mild NPDR at index, eyes diagnosed with severe NPDR and PDR were 3.6 and 4.0 times more likely to develop SB within 2 years, respectively. Factors associated with increased risk of developing SB included black (hazard ratio (HR), 1.24; P < 0.0001) and Asian race (1.27; P < 0.011), female sex (HR 1.08; P = 0.01), and enrollment in Medicare or Medicaid compared with private plans (HR 0.70-0.91; P < 0.05), in addition to diabetic macular edema and retinal vein occlusion during follow-up (HR 1.96-4.91; P < 0.0001).

Conclusions: Our model identified that race, sex, and other characteristics can predict likelihood of developing SB in patients with DR. Close monitoring or early intervention should be considered for patients who are black or Asian race, female, or have developed other ocular conditions.

Disclosure

Q. Nguyen: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Genentech, Inc., Regeneron Pharmaceuticals. R. Hall: Employee; Self; American Academy of Ophthalmology. S.P. Kelly: None. F. Lum: None. I. Stoilov: Employee; Self; Genentech, Inc. I.M. Abbass: Employee; Self; Genentech, Inc. T. To: Employee; Self; Genentech, Inc. Stock/Shareholder; Self; Genentech, Inc. A. Abolian: Employee; Self; Genentech, Inc. V. Garmo: Employee; Self; Genentech, Inc.
500-P: The Performance of Four Different Exosome Isolation Methods for Urinary Exosomal microRNA Profiling in Patients with Diabetic Kidney Disease
Authors: LEE KIYOUNG , LEE DAEHO , PARK IE BYUNG
Published: June 2019

Both exosome and microRNAs (miRNAs) in biological fluids can be good biomarkers for pathologic conditions. Thus, urinary exosome miRNAs would be reasonable non-invasive tools for biomarker development for diabetic kidney disease. To be eligible for exosome miRNA biomarker development, the performance of exosome isolation methods for miRNA profiling need to be tested. In the present study, using urine samples from patients with type 2 diabetes, we compared four different exosome isolation methods for their performance in urine exosome isolation and detectable urinary exosome miRNA profiles. Four methods compared in the present study were ultracentrifugation (UC), size exclusion chromatography using qEV column (qEV), ExoQuick-TC (ExoQuick), and Amicon tube 100K (Amicon). Isolated exosomes were subjected to next generation sequencing (NGS) to profile miRNAs. The counts, RNA concentrations, and protein marker expressions of isolated urine exosome were variable between the extraction methods, with the highest exosome count with the qEV method. The number of miRNAs detected (read count ≥1) in 100 mL urine each were 314, 174, 91, and 130 from UC, qEV, ExoQuicK, and Amiconmethods, respectivley. And, 57 miRNAs were detected with all 4 methods concordantly. There were correlations in the levels of exosomal miRNAs between samples from UC, qEV, and ExoQuick methods, whereas exosome miRNA profiles from Amicon sample showed only a weak correlation with those from other methods. Our results suggest that exosome isolation by using UC or qEV method may be a better option among different isolation methods to profile urinary exosome miRNAs in diabetic kidney disease. However, ultrafiltration method using Amicon tube produced a different profile of miRNAs with other methods. Further studies are needed to characterize both exosome and their genetic materials from different extraction sources.

Disclosure

K. Lee: None. D. Lee: None. I. Park: None.
505-P: ROCK2 Regulates TGF-Beta-Induced Expression of CTGF and Profibrotic Genes via NF-kappa B and Cytoskeleton Dynamics in the Mesangial Cells
Authors: NAGAI YOSUKE , KAWANAMI DAIJI , MATOBA KEIICHIRO , TAKEDA YUSUKE , UTSUNOMIYA KAZUNORI
Published: June 2019

The small GTPase Rho and its effector Rho-kinase are involved in the pathogenesis of diabetic kidney disease. Accumulating evidence shows profibrotic mediators, including connective tissue growth factor (CTGF), are key regulators of glomerulosclerosis under diabetic conditions. However, the interactions of Rho-kinase and these profibrotic mediators in the development of renal dysfunction remain unclear. Glomeruli isolated from type 2 diabetic db/db mice demonstrated increased gene expression of transforming growth factor β (TGF-β) and its downstream profibrotic mediators. Chemical inhibition of Rho-kinase suppressed the expression of profibrotic mediators in both isolated glomeruli and cultured mesangial cells. An investigation of mechanisms underlying this observation revealed activated Rho-kinase functions through the phosphorylation of JNK and Erk, and the nuclear translocation of nuclear factor-kappa B (NF-κB). Immunostaining studies demonstrated that a Rho-kinase inhibitor and Cytochalasin D, an inhibitor of actin polymerization, attenuated TGF-β-mediated nuclear translocation of NF-κB, indicating that Rho-kinase-mediated actin reorganization regulates NF-κB activation. Knockdown by small interfering RNA against two Rho-kinase isoforms, ROCK1 and ROCK2, showed that ROCK2 but not ROCK1 controls this fibrotic machinery. SLx-2119, a chemical ROCK2 inhibitor, also modulated the expression of TGF-β-induced profibrotic mediators via a similar mechanism by downregulating ROCK2 activity. A further in vivo study showed that ROCK2 activity in the renal cortex of db/db mice was elevated compared to control db/m mice. These observations indicate that ROCK2 is a key player in the development of diabetic renal injury. Glomerular ROCK2 may be a potential therapeutic target for the treatment of diabetic kidney disease.

Disclosure

Y. Nagai: None. D. Kawanami: Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc. K. Matoba: Research Support; Self; Daiichi Sankyo Company, Limited, Novo Nordisk Inc., Sanofi, Shionogi & Co., Ltd., Takeda Pharmaceutical Company Limited. Y. Takeda: None. K. Utsunomiya: None.
512-P: Sexual Differences in Progression of Diabetic Nephropathy in over 26 Type 1 Diabetic Mice
Authors: WANG WANNING , CAI LU
Published: June 2019

As a transgenic type 1 diabetic model, OVE26 mice develop diabetic nephropathy (DN) with the feature close to advanced DN in human, which makes it a valuable rodent model to study DN. Nevertheless, sexual differences in renal lesions, especially dynamic progression with different ages, remain undefined in OVE26 mice. To explore these issues, male and female OVE26 and wild type nondiabetic FVB mice at 4-, 12-, 24 - and 36-weeks old were used (N=6). Urine albumin-to-creatinine ratio (UACR) was detected to evaluate renal function. Renal histopathology and immunochemical and Western blotting protein expression of inflammation, tubular epithelial-myofibroblast transdifferentiation and fibrosis markers were examined. In FVB mice, blood glucose, triglyceride, UACR and podocytes numbers were constant during aging and comparable in male and female. Age-dependent slightly increase of glomerular mesangial matrix, inflammatory cells infiltration and renal fibrosis were observed in FVB mice. Compared with the age- and sex- matched FVB mice, male OVE26 mice developed hyperglycemia, mesangial matrix accumulation and increased trend of triglyceride at 4-week-old and albuminuria, glomerular enlargement, and upregulations of TGF-β1, α-SMA at 12 weeks old. Significant loss of podocytes, tubular dilation, collagen accumulation, increased trend of macrophage and T lymphocytes infiltrations, and increased protein levels of TNF-α, ICAM and fibronectin in kidney were observed in 24-week old male OVE26 mice. Compared with age-matched male OVE26 mice, female OVE26 mice had significant hypertriglyceridemia, worse renal function and kidney lesions at older age, including higher UACR and sustained loss of podocytes at age of 24 weeks; more inflammatory cells infiltration and renal fibrosis at age of 36 weeks. Our findings suggest the sexual difference in the progression of DN in OVE26 mice to provide evidence for researchers to properly select the age and sex of OVE26 mice in the future type 1 DN study.

Disclosure

W. Wang: None. L. Cai: None.
514-P: Group 2 Innate Lymphoid Cells Promote Renal Fibrosis through the Production of Th2 Cytokines in Diabetic Kidney Disease
Authors: QIN LU DAN , LIU CUIPING , ZHOU LUPING , GAO CHENLIN , ZHANG TING , GUO MAN , HUANG WEI , JIANG ZONGZHE , DING JING YA , XU YONG
Published: June 2019

Aim: To explore the role of Group 2 innate lymphoid cells (ILC2s) in the pathogenesis of renal fibrosis in diabetic kidney disease (DKD).

Methods: The proportion of ILC2s and ILC2s-producted Th2 cytokines (IL-4, IL-5, IL-13) in the peripheral blood of normal control subjects (NC) or patients with type 2 diabetes (DM), early diabetic kidney disease (DKD1), or late diabetic kidney disease (DKD2) were analyzed using flow cytometry and ELISA. Renal tubular epithelial cells (HK-2) were stimulated with IL-4 or IL-13 in the presence or absence of high glucose and the expression and release of transforming growth factor-β1 (TGF-β1) and fibronectin (FN) was analyzed using qPCR and ELISA.

Results: The proportion of ILC2s and the level of IL-4, IL-5, and IL-13 were significantly increased with the severity of DKD (P<0.05). The expression and release of TGF-β1 and FN were significantly upregulated by IL-4 or IL-13 in HK-2 cells (P<0.05). Compared with high glucose stimulation alone, treatment with IL-4 or IL-13 combined with high glucose significantly stimulated HK-2 cells to increase expression of FN and TGF-β1 (P<0.05).

Conclusions: ILC2s may promote the development and progression of diabetic kidney disease by secreting Th2 cytokines (IL-4 and IL-13) and promoting renal fibrosis.

Disclosure

C. Liu: None. L. Zhou: None. C. Gao: None. T. Zhang: None. M. Guo: None. W. Huang: None. Z. Jiang: None. J.Y. Ding: None. Y. Xu: None.
530-P: Early Insulin Initiation in T2DM Is Associated with Prevention of eGFR Decline and Lower Insulin Dose Requirements
Authors: KESAVADEV JOTHYDEV , WARRIER ROHIT , RAMACHANDRAN LAKSHMY , SHANKAR ARUN , DAVID ASHWIN , AJAI NEETHU A. , SANAL GEETHU , THAMPIRAJ KAVITHA , KRISHNAN GOPIKA , JOTHYDEV SUNITHA
Published: June 2019

Early initiation of insulin therapy often remains contentious due to various patient and physician-related barriers. Insulin is delayed by several years despite high A1c and is the foremost reason for costly future complications. In addition, the problem of early aggressive use of insulin depleting drugs causes β-cell fatigue leading to "brittle diabetes" and higher insulin dose requirements. In this observational cohort study, a comparison has been made between early insulin initiators (EII) (within 7-8 years of diabetes onset) and late insulin initiators (LII) (>8 years of onset), in terms of progression of CKD (eGFR decline), if any. A total of 200 subjects in each arm were matched at baseline for T2DM duration (16.78±4.32 year), BP, BMI, HbA1c and presence of co-morbid illnesses. Both groups were closely followed-up through our Diabetes Tele Management System based health care model that have previously shown to be successful in safely achieving glycemic targets compared to conventional models of care. A paired t test was done for analyzing the eGFR decline within each arm. EII arm required a significantly lower total daily dose of insulin (TDD) (average of 9 units/day) to maintain glycemic control throughout than the LII arm (average of 30 units/day). The decline in eGFR was significant in the LII arm, whereas in the EII arm, eGFR was maintained throughout. Change in A1c was also significant.

Disclosure

J. Kesavadev: None. R. Warrier: None. L. Ramachandran: None. A. Shankar: None. A. David: None. N.A. Ajai: None. G. Sanal: None. K. Thampiraj: None. G. Krishnan: None. S. Jothydev: None.
532-P: Effect of ABC Control of Diabetes on the Prevalence of Chronic Kidney Disease in Patients with Type 2 Diabetes from Tibet in China
Authors: CHENGHUI ZHANG , YUNHONG WU , SUYUAN WANG , MINGXIA LI
Published: June 2019

Aim: To investigate the effect of ABC control of diabetes (A: HbA1c, B: blood pressure and C: LDL-C) on the prevalence of chronic kidney disease (CKD) in patients with type 2 diabetes (T2D) from Tibet based on electronic medical records.

Methods: Patients (aged 18-79 years) documented from 2014 to 2018 were enrolled. Control criteria were defined base on the 2017 Chinese Guidelines for the Prevention and Treatment of Type 2 Diabetes: HbA1c < 7.0%, blood pressure <130/80 mmHg and LDL-C <1.8 mmol/L with coronary heart disease (CHD) and 2.6 mmol/L without CHD. CKD was defined as eGFR <60 ml/min per 1.73 m2 and/or urinary albumin/creatine ratio ≥ 30 mg/g. Patients were divided into four groups according to ABC control.

Results: A total of 1999 patients were enrolled. The mean (SD) age was 55.2 (10.5) years. The median diabetic duration was 7.0 (range: 0.1-32) years and 43.9% patients were female. As shown in Figure 1, the prevalence of CKD has increased in parallel with the bad control of ABC (P<0.001). Compared with patients achieved all three targets, patients with none of target achieved had a 2.70-fold (95% CI: 1.25-5.86, P = 0.012) higher risk for CKD after adjusted for age, gender and diabetic duration.

Conclusions: The achievement of ABC target was associated with the prevalence of CKD. More intensive T2D management is warranted to prevent the development of CKD in this population.

Disclosure

Z. Chenghui: None. W. Yunhong: None. W. Suyuan: None. L. Mingxia: None.
534-P: Serum Sclerostin, Body Composition, and Sarcopenia in Hemodialysis Patients with Diabetes
Authors: BANDEIRA FRANCISCO , MEDEIROS MARIA CAROLINA W. , JUNIOR MARIO HENRIQUES , SILVA NATALIA ROCHA , BANDEIRA FARIAS MARIA ELBA , FARIAS LEONARDO
Published: June 2019

Sclerostin (Scl) is an osteoblast inhibiting glycoprotein secreted mainly by osteocytes which is regulated by hormonal changes and skeletal loading. Decreased physical function and high serum Scl concentrations have been reported in chronic renal failure patients but little is known about the differences among diabetic and nondiabetic patients on hemodyalisis who are susceptible to both sarcopenia and bone fragility.

Objective: To determine the prevalence of sarcopenia and its association with serum sclerostin concentrations and metabolic parameters in 92 patients on hemodyalisis. Anthropometric data and physical performance were evaluated. Blood sample were collected for sclerostin, glucose, cholesterol, triglycerides, calcium, phosphate, PTH e 25OH-vitamin D measurements. Lean mass was evaluated using multifrequency electro-bioimpedance at a “dry weight” time (after dialysis session).

Results: Mean age was 63.3±13.6 years, 63% were male and 44.6% had diabetes. Mean BMI was higher in diabetics (26.6±5.2 vs. 24.1±3.7; p=0.01) and there were no differences in gait speed and hand grip strength between diabetic and nondiabetic subjects. A low skeletal muscle mass index (SMI) was identified in 65.2% of the participants, and among them 76.7% were men and 36.7% diabetics. Mean serum sclerostin was 86.9 ± 39.0 pmol/L, which was higher in men (94.6 ± 41.7; p = 0.017), in individuals with low SMI (94.9 ± 40.7; p < 0.001) and in diabetics (97.2 ± 46.6; p < 0.003). After multivariate analysis and adjustments for potential confounders, high serum sclerostin was independently associated with low SMI and with the presence of diabetes. The following variables correlated positively with diabetes: blood pressure, body mass index (BMI) waist circumference, waist/hip ratio, plasma glucose, serum sclerostin and fat mass.

Conclusions: We found higher serum sclerostin concentrations in hemodialysis patients with diabetes which were inversely related to muscle mass.

Disclosure

F. Bandeira: None. M.W. Medeiros: None. M.H. Junior: None. N.R. Silva: None. M. Bandeira Farias: None. L. Farias: None.

Funding

Arab Society for Paediatric Endocrinology and Diabetes
537-P: sCD93 Is a Potential Biomarker for Diabetic Nephropathy
Authors: LEE SANG BAE , PARK HO SEON , LEE MINYOUNG , CHOI ARIM , PARK JONG SUK , AHN CHUL WOO , LEE SEUNG-PYO , KANG SHINAE
Published: June 2019

Objective: CD93 is a glycoprotein that is known to enhance phagocytosis. The extracellular portion of CD93 can be enzymatically cleaved and secreted to the serum as a soluble form (sCD93). Diabetic nephropathy (DN) is a well-known inflammatory disease that commonly progresses to end stage renal disease (ESRD) in diabetes. Although albuminuria and estimated glomerular filtration rate (eGFR) is currently used for the diagnosis and the prognostication of DN, the prognostic value of the two parameters is modest at best. Here, we hypothesized that sCD93 would be a new biomarker for DN.

Methods: We registered 97 human subjects with type 2 diabetes and evaluated the association between serum sCD93 and DN. The underlying mechanism of CD93 regulation was elucidated using human umbilical cord endothelial cell (HUVEC) in vitro and diabetic db/db mice in vivo.

Results: The group with higher sCD93 has lower eGFR level and higher prevalence of CKD defined by eGFR. The log(sCD93) was significantly associated with albumin-to-creatinine ratio (ACR) (r=0.229, p=0.028) and the sCD93 level was an independent determinant of ACR and eGFR. The odds ratio of having DN was higher in the high serum sCD93 group even after adjusting for other covariates (adjusted odds ratio 7.283, 95% CI 1.255-42.268, p=0.027). In vitro, CD93 was highly expressed in HUVEC and both CD93 expression and sCD93 secretion was upregulated after LPS stimulation. In db/db mice, sCD93 level was much higher in the peritoneal fluid compared to the db/m mice. The db/db mice showed significantly higher CD93 expression in the renal glomerulus compared to that of the db/m mice suggesting that the elevated sCD93 according to the degree of DN progression might be associated with upregulated expression of CD93 in glomerulus.

Conclusion: Based on these human and rodent data, we conclude that sCD93 could be a candidate molecule associated with DN.

Disclosure

S. Lee: None. H. Park: None. M. Lee: None. A. Choi: None. J. Park: None. C. Ahn: None. S. Lee: None. S. Kang: None.

Funding

Korean National Research Foundation (2014R1A1A3051221)
543-P: A Clinical, Proteomics and Artificial Intelligence-Driven Model to Predict Acute Kidney Injury in Diabetic Patients Undergoing Coronary Angiography—Results from the Catheter Sampled Blood Archive in Cardiovascular Diseases Study
Authors: IBRAHIM NASRIEN E. , MCCARTHY CIAN P. , SHRESTHA SHREYA , GAGGIN HANNA , MUKAI RENATA , MAGARET CRAIG A. , RHYNE RHONDA F. , JANUZZI JAMES
Published: June 2019

Background: We previously developed a clinical/proteomics panel for predicting procedural acute kidney injury (AKI) and now examine its performance in those with diabetes (DM).

Methods: We measured 109 biomarkers in blood from patients undergoing coronary angiography. Procedural AKI defined as increase in serum creatinine ≥0.3 mg/dL, increase in serum creatinine of ≥50%, or documented oliguria ≤7 days after procedure. Clinical and biomarker predictors of AKI identified using least-angle regression; a final prognostic model was developed with LASSO; the model was then measured in those with DM.

Results: Besides DM, 5 predictors were in the final model: 3 (blood urea nitrogen to creatinine ratio, C-reactive protein and osteopontin) had positive association, while 2 (CD5 antigen-like and Factor VII) had negative association with AKI risk. Among 217 patients with DM, 18 (8.3%) developed AKI. The final model had an AUC of 0.87 for predicting procedural AKI (P<0.001). The optimal score cut-off had 100% sensitivity and a negative predictive value (NPV) of 100% for procedural AKI.

Conclusions: We describe a clinical and proteomics-supported biomarker model with high sensitivity/NPV for AKI in patients with DM following coronary angiography.

Disclosure

N.E. Ibrahim: Other Relationship; Self; Novartis Pharmaceuticals Corporation. C.P. McCarthy: None. S. Shrestha: None. H. Gaggin: Other Relationship; Self; Dr. Gaggin has received research grant support from Roche Diagnostics, Jana Care, Ortho Clinical, Novartis; consulting income from Merck, Roche Diagnostics; research payments for clinical endpoint com. R. Mukai: None. C.A. Magaret: Consultant; Self; Prevencio. R.F. Rhyne: Board Member; Self; Prevencio. Employee; Self; Prevencio. Stock/Shareholder; Self; Prevencio. J. Januzzi: Consultant; Self; Abbott, Roche Diagnostic USA. Research Support; Self; Abbott, Prevencio, Quest Diagnostics, Roche Diagnostic USA.

Funding

Prevencio, Inc.