bluuebunny/crossref_metadata_2025_split huggingface dataset:
456-P: Targeted Deletion of Insulin Receptor in Vascular Smooth Muscle Cells Accelerated Thinning of Plaque Cap and Apoptosis and Atherosclerosis
Authors: LI QIAN , FU JIALIN , PARK KYOUNGMIN , ZAITSU YUMI , ST-LOUIS RONALD , RASK-MADSEN CHRISTIAN , KING GEORGE L. 
Published: June 2020

Vascular smooth muscle cells (VSMCs) play an important role in the development of the stability of atherosclerotic plaque. We reported that knockout of insulin receptor (IR), but not IGF1 receptor, in VSMCs reduced intimal hyperplasia of femoral artery after wire injury. To investigate the role of IR in VSMCs on atherosclerotic plaque, ApoE and smooth muscle cells IR double knockout (SMIRKO/ApoE-/-) mice were generated. Extent of lipid deposition and atherosclerosis were increased in the aorta of SMIRKO/ApoE-/- mice compared to control mice (p&lt;0.05). However, total cell numbers and proliferation of VSMCs, cap thickness and collagen of the plaque were decreased, whereas VSMCs apoptosis, macrophage content and necrotic area were increased in the aortic plaques of SMIRKO/ApoE-/- mice compared to ApoE-/- mice. VSMCs cultured from SMIRKO/ApoE-/- mice exhibited more apoptosis by withdrawing growth factors which were inhibited by insulin compared to VSMCs from ApoE-/- mice. Analysis of genes in VSMCs subsets which were reported to be increased in aortic plaques showed MMP2, decorin, thrombospondin 1 and 5, sFRP3 and IGFBP4 were increased in the aortic media of SMIRKO/ApoE-/- mice vs. ApoE-/- mice, with MMP2, thrombospondin 1 and 5 and sFRP3 were also increased in the aortic media of mice fed with high fat diet or diabetic mice due to streptozotocin injection. Insulin downregulated MMP2 and thrombospondin 1 in cultured VSMCs, whereas overexpression of Foxo1 in VSMCs increased the expression of thrombospondin 1. Reanalysis of existed microarray data base and found MMP2 and thrombospondin 1 were also elevated in internal mammary artery of diabetic compared to nondiabetic people. Thus, insulin receptor action on VSMCs can change expression of extracellular genes and its turnover to increase the stability of atherosclerotic plaque.

Disclosure
Q. Li: None. J. Fu: None. K. Park: None. Y. Zaitsu: Employee; Self; Sunstar Group. R. St-Louis: None. C. Rask-Madsen: None. G.L. King: Research Support; Self; Janssen Pharmaceuticals, Inc.

Funding
National Institutes of Health (R01DK053105)
468-P: PDK1 Is a Key Regulator of Podocyte Viability and Maintenance of Glomerular Function
Authors: IWATA WAKIKO , UNOKI-KUBOTA HIROYUKI , KATO HIDEKI , MATSUMOTO MICHIHIRO , NANGAKU MASAOMI , KABURAGI YASUSHI 
Published: June 2020

In diabetic states, altered activity of insulin signaling pathway is likely to play key roles in insulin resistance and in kidney dysfunction. Since the 3-phosphoinositide-dependent protein kinase 1 (PDK1) serves as a major signaling molecule in the regulation of cellular responses in multiple organs by mediating the phosphoinositide 3-kinase signaling pathway, we tested the hypothesis that podocyte-specific deletion of PDK1 leads to podocyte injury and proteinuric kidney disease. PDK1 was selectively silenced in podocytes using Cre-mediated recombination controlled by the podocin promoter. Mice with a podocyte-specific deletion of PDK1 (PDK1 KO) developed proteinuria, and died due to end-stage renal disease by 5 weeks after birth. Histologically, features characteristic of glomerulosclerosis, including podocyte foot process effacement, mesangial sclerosis, and protein casts, were observed in PDK1 KO at 3 weeks of age. We also examined podocyte distribution in glomeruli using the podocyte markers synaptopodin and WT-1, and found that their expressions were lost in some glomeruli of PDK1 KO at 3 weeks of age although their expression patterns at 1 weeks of age PDK1 KO were almost comparable with control mice. We next examined a possible role of forkhead transcription factor, forkhead box O1 (FoxO1) in linking PDK1 of podocytes to kidney function. For this, we generated podocyte-specific PDK1 and FoxO1 double knockout mice (DKO) by introducing Foxo1-flox gene into PDK1 KO mice. DKO exhibited a reduced level of proteinuria compared with age-matched PDK1 KO, and the maximal survival time of DKO had extended to 6.6 weeks after birth. Taken together, PDK1-FoxO1 pathway may be one of the critical signal transduction pathways that links podocyte injury and kidney dysfunction in diabetic states.

Disclosure
W. Iwata: None. H. Unoki-Kubota: None. H. Kato: None. M. Matsumoto: None. M. Nangaku: Consultant; Self; Akebia Therapeutics, Astellas Pharma Inc., Bayer AG, Boehringer Ingelheim International GmbH, Daiichi Sankyo, GlaxoSmithKline plc., Japan Tobacco Inc., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Novo Nordisk Inc., Otsuka Pharmaceutical Co., Ltd., Scohia Pharma Inc. Research Support; Self; Takeda Pharmaceutical Company Limited. Speaker’s Bureau; Self; AstraZeneca, Torii Pharmaceutical Co. Ltd. Y. Kaburagi: None.
469-P: ROCK1/AMPK Axis Regulates the Development of Diabetic Kidney Disease via Modulation of Fatty Acid Metabolism
Authors: NAGAI YOSUKE , MATOBA KEIICHIRO , TAKEDA YUSUKE , AKAMINE TOMOYO , KAWANAMI DAIJI , UTSUNOMIYA KAZUNORI , NISHIMURA RIMEI 
Published: June 2020

The small GTPase Rho and its effector Rho-kinase are involved in the pathogenesis of diabetic kidney disease. Accumulating evidence shows that renal dysfunction in diabetic patient is associated with abnormal fatty acid oxidation in the kidney. However, the interaction of Rho-kinase and fatty acid oxidation in diabetic kidney remain unclear. Glomeruli isolated from type 2 diabetic db/db mice demonstrated decreased gene expression of fatty acid oxidation mediators such as peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), CD36, carnitine palmitoyltransferase 1A (CPT1A). Chemical inhibition of Rho-kinase restored expression of fatty acid oxidation-related genes in both isolated glomeruli and cultured mesangial cells. An investigation of mechanisms underlying this observation revealed activated Rho-kinase functions through the phosphorylation of AMPK and increased expression of PGC-1α. Extracellular flux analyzer demonstrated that Rho-kinase inhibition improves TGF-β-induced mitochondrial dysfunction. Furthermore, fluorescence histochemistry showed that Rho-kinase inhibitors suppresses ROS production as a result of mitochondrial damage due to abnormal fatty acid metabolism. Knockdown by small interfering RNA against two Rho-kinase isoforms, ROCK1 and ROCK2, showed that ROCK1 but not ROCK2 controls this metabolic machinery. Consistent with this result, mesangial cells isolated from ROCK1 deficient mice were protected from TGF-β-mediated downregulation of fatty acid metabolism. These observations indicate that ROCK1 is a key player in the development of diabetic renal injury. Glomerular ROCK1 may be a potential therapeutic target for the treatment of diabetic kidney disease.

Disclosure
Y. Nagai: None. K. Matoba: Research Support; Self; Sanofi K.K., Takeda Pharmaceutical Company Limited. Y. Takeda: None. T. Akamine: 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.
473-P: HDAC Inhibition Augments TGF-ß-Induced JNK Activation, CTGF Expression, and NF-E2 Degradation Promoting Profibrotic Signaling in Renal Proximal Tubules
Authors: LI JIA , RANE MADHAVI JAGDISH , CAI LU , TAN YI 
Published: June 2020

The role of TGF-β in diabetes-induced renal fibrosis remains incompletely understood. We showed decreased NFE2 expression in TGFβ-treated renal proximal tubule cells and in diabetic mouse kidneys. TGF-β treatment increased NFE2 mRNA level in HK11 cells suggesting a role for NFE2 degradation. ERK and p38 MAPKs promoted TGFβ-induced NFE2 degradation, CTGF and FN expression. NFE2 over-expression/silencing inhibited exacerbated TGFβ-induced CTGF. Proteasome inhibitor MG132 preserved TGFβ-induced NFE2 expression and inhibited CTGF and FN expression. Current study examined role of JNK MAPK and HDAC inhibition on NFE2 and CTGF expression in TGF-β treated HK-11 cells and effects of MG132 on JNK activation in OVE26 mouse kidneys and in TGFβ-treated HK-11 cells.HK11 cells were pre-treated with/without JNK inhibitor SP600125, HDAC inhibitor trichostatin A, and MG132, for an hour prior to treatment with TGFβ for 24 h.JNK blockade inhibited TGFβ-induced CTGF expression but not NF-E2 expression. MG132 blocked TGFβ-induced JNK activation in HK11 cells and in OVE26 mice. HDAC inhibition exacerbated TGFβ-induced NFE2 degradation and induced CTGF expression. ERK, p38 and JNK pathways contribute to enhanced CTGF expression but only p38 and ERK MAPK pathways contribute to NFE2 degradation. MG132 blocked TGFβ-induced JNK activation, suggesting that proteasome function is necessary for JNK activation and therefore, blocking JNK had no effect on preserving NF-E2 expression. Blocking HDAC activation possibly promoted HAT activity, NFE2 acetylation and enhanced degradation. Thus, proteasome inhibition plus TGFβ preserved NFE2 expression and prevented JNK activation and CTGF. HDAC inhibition plus TGFβ exacerbated NFE2 degradation and enhanced JNK activation and CTGF. We identified NFE2 as a potential anti-fibrotic protein in the kidney.

Disclosure
J. Li: None. M. Rane: None. L. Cai: None. Y. Tan: None.
474-P: Gut Microbiome–Derived Phenyl Sulfate Contributes to Albuminuria in Diabetic Kidney Disease
Authors: KIKUCHI KOICHI , ABE TAKAAKI 
Published: June 2020

Diabetic kidney disease is a major cause of renal failure in urgent of breakthrough in disease management. Type 2 diabetes causes significant changes in an array of plasma metabolites, and in the humans, SLCO4C1 is the only transporter contributes to transport them into urine. We generated transgenic (Tg) rats overexpressing SLCO4C1 in the proximal tubule, a typical model for human renal excretion. Using this model, we characterize metabolites that are increased in diabetic wild type rats, but reduced in diabetic Tg rats.PS increase with the progression of diabetes and was decreased in Tg rats with limited proteinuria. In diabetes models, PS administration induced albuminuria and podocyte damage due to the mitochondrial dysfunction. Phenol is synthesized from tyrosine by gut bacterial-specific tyrosine phenol-lyase (TPL) and phenol is metabolized to PS in the liver. Administration of TPL inhibitor reduced not only serum PS level but also albuminuria in diabetic mice. Furthermore, TPL inhibitor significantly ameliorate renal dysfunction in adenine-induced renal failure model. Because TPL inhibitor did not alter the major composition, the non-lethal inhibition of microbial-specific enzymes has a therapeutic advantage with lower selective pressure for the development of drug resistance. We next confirmed whether PS also related with the diabetes and DKD progression in humans. In a diabetic patient cohort (U-CARE study, n=362), serum PS levels related with the basal albuminuria level. In addition, logistic regression analysis showed that among known ACR predictive factors, PS was the only factor which significantly related 2-year progression of albuminuria in patients with microalbuminuria. These data suggested that PS may have a potential as important predictive marker of DKD.PS is a modifiable cause and a target for the treatment of DKD. Chemical reduction of TPL should represent another aspect for developing drugs preventing DKD (Kikuchi et al. Nat. Commun. 10: 1835, 2019).

Disclosure
K. Kikuchi: None. T. Abe: None.
484-P: Abnormal Ketone Body Metabolism Caused by Bdh1 Deficiency Promoted the Progression of Diabetic Kidney Disease by Activating ROS
Authors: WAN SHENGRONG , TENG FANGYUAN , JIANG ZONGZHE , XU YONG 
Published: June 2020

Objective: This study was intended to reveal the mechanism of ketone body metabolism abnormality caused by bdh1 deficiency promoting the progression of diabetic kidney disease(DKD) by activating reactive oxygen species (ROS).
Methods: Eight-week-old wild type(n=8) and db/db (n=8) mice were treated with normal diet and high-fat diet for three months, respectively. Then two groups of mice kidney samples were taken for RNA-Seq analysis to select the target gene. It was verified by qRT-PCR, Western blot, immunofluorescence, and immunohistochemistry in mice kidney. A high glucose-induced ROS activation model of HK-2 renal cells was established, and then siRNA was used to interfere with bdh1.
Results: RNA-seq showed that the pathway of ketone body metabolism in DKD changed significantly, and further analysis showed that bdh1, the key protein of ketone body metabolism, decreased significantly. The sequencing result was further confirmed by qRT-PCR, Western blot, immunofluorescence, and immunohistochemistry, and we found that bdh1 was mainly expressed in tubular epithelial cells. In order to identify the molecular mechanism of bdh1 involved in DKD, we constructed the high glucose-induced ROS activation model of HK-2 cells in vitro. It was found that bdh1 was significantly down-regulated in high glucose treated HK-2 cells. The knockdown of bdh1 by siRNA could promote the production of ROS induced by high glucose. Further research found that bdh1 promoted the expression of nrf2 then inhibited ROS through the ketone body metabolism pathway, thus protected HK-2 cells in a high glucose environment.
Conclusions: Bdh1 promoted the expression of nrf2 through the pathway of ketone body metabolism and inhibited the production of ROS to protect HK-2 cells in a high glucose environment. Abnormal ketone body metabolism caused by bdh1 deficiency participated in the progress of DKD.

Disclosure
S. Wan: None. F. Teng: None. Z. Jiang: None. Y. Xu: None.
49-OR: Synergistic Effect of ERAD and Autophagy in Preventing Proinsulin Aggregation and Promoting Beta-Cell Survival
Authors: SHRESTHA NEHA , REINERT RACHEL B. , ARVAN PETER , QI LING 
Published: June 2020

Decline of proteostasis in the endoplasmic reticulum (ER) is thought to underlie molecular changes in β cell function and drive the pathogenesis of diabetes. ER-associated degradation (ERAD) and macroautophagy (hereafter “autophagy”) are two principal quality-control mechanisms responsible for the clearance of ER-resident proteins or protein aggregates, respectively, in the cell. However, the physiological significance of ERAD-autophagy crosstalk remains unexplored. Previously, using β cell-specific knockout mouse models, we reported that ERAD deficiency leads to β cell dedifferentiation, while autophagy deficiency leads to β cell loss by 4-5 months of age. Interestingly, we observed that loss of ERAD in β cells triggered activation of autophagy pathways, as monitored by LC3 flux. Furthermore, chloroquine-mediated inhibition of autophagy in ERAD-deficient islets led to an increased proinsulin aggregation. In mice, combined deletion of ERAD and autophagy in β cells caused marked hyperglycemia soon after weaning, with extensive loss of β cells and premature death at 2-3 months of age. Thus, our data demonstrate that ERAD and autophagy act synergistically to prevent proinsulin aggregation and promote β cell survival. This study reveals a critical role for crosstalk between the ERAD and autophagy pathways in preventing diabetes and provides the first evidence for synergism between ERAD and autophagy in vivo.

Disclosure
N. Shrestha: None. R.B. Reinert: None. P. Arvan: None. L. Qi: None.

Funding
JDRF (2-SRA-2018-539-A-B); National Institutes of Health (1R01DK117639-01, 1R01DK111174)
500-P: Renal Function Decline and the Related Risk Factors in a Cohort of Beijing Community Diabetes Study
Authors: YUAN MINGXIA , YUAN SHENYUAN ,   BEIJING COMMUNITY DIABETES STUDY (BCDS) GROUP
Published: June 2020

Objective: To explore the renal function decline and the related risk factors in a cohort of Beijing Community Diabetes Study.
Methods: A multi-center, prospective study with ten-year follow-up was conducted at Beijing communities between 2008 and 2018. The clinical information and physical examination were recorded. Laboratory test including hemoglobin A1c, serum uric acid, fasting blood glucose, serum creatinine, and blood lipid profile were conducted. The patients were divided into two groups according to their age (&lt;65 and ≧ 65). Annual eGFR decline rate≧3ml/min/1.73m2 was defined as rapid renal function decline. The tendency of renal function change over 10 years in different age groups had been described. Logistic multivariable models were employed to investigate the risk factors of rapid renal function decline.
Results: A total of 1655 patients with type 2 diabetes were enrolled. The median (quartile1, quartile3) annual rate of decline in eGFR of T2DM was 0.21(-2.51, 3.06) ml/min/1.73m2. Among them, 25.5% of the patients (n=422) showed rapid GFR decline, with an annual decrease of 5.28(3.85, 7.46) ml/min/1.73m2. Non-rapid decliner was 74.5% (n=1233), and the annual decrease was -0.99(-3.66, 0.82) ml/min/1.73m2. The patients with end-stage renal disease increased from 0.12% to 0.91% compared with the baseline. Uric acid was an independent risk factor for rapid GFR decline in both non-elder (OR=1.01, 95%CI: 1.00-1.01, P=0.001) and elder population (OR=1.01, 95%CI: 1.00-1.01, P=0.002), adjusting for age, sex and duration. Hyperfiltration was another independent risk factor for rapid GFR decline in non-elder (OR=14.72, 95%CI: 8.97-24.15, P&lt;0.001) and elder population (OR=7.91, 95%CI: 4.04-15.48, P&lt;0.001).
Conclusions: A quarter of patients with type 2 diabetes showed rapid renal function decline over 10 years. Serum uric acid and glomerular hyperfiltration are independent risk factors to rapid renal function decline.

Disclosure
M. Yuan: None. S. Yuan: None.

Funding
Capital Health Research and Development (2016-1-2057)
502-P: Tubular Markers and Diabetic Retinopathy Can Predict Early Diabetic Kidney Disease
Authors: ZHANG LIN , LU JING 
Published: June 2020

Aim: To evaluate the association of tubular markers and diabetic retinopathy (DR) in 2-DM patients with normal creatinine(Cr) and urinary microalbumin/Cr(ACR).
Methods: In this study, we selected 1164 patients with 2-DM to integrate the clinical biochemical indexes and urinary alpha-1 microglobulin/Cr(A1CR), urinary immunoglobulin G/Cr(IGUCR), urinary transferrin/Cr(TRUCR), ACR and fundus examination. 274 patients underwent nuclear medicine 99mTc-EC to measure renal effective plasma flow (ERPF) and 99mTc-DTPA to measure glomerular filtration rate (GFR). They were divided into 2 groups: 52 cases with DR and 222 cases non-DR. Renal parameters were compared between 2 groups.
Results: In this cross-sectional study, we fund the tubular markers: A1CR, IGUCR, and TRUCR in the DR group were higher (P &lt; 0.001) than non-DR group, while ERPF decreased (P &lt; 0.001). The glomerular parameters: ACR was slightly higher in the DR group, GFR had no significant difference between 2 groups (P &gt; 0.05). Pearson correlation analysis showed A1CR had a better correlation with GFR and ERPF (P &lt; 0.001) than ACR (P &gt; 0.05). In a binary logistic analysis, after adjusting sex, age, course of DM, and other factors, A1CR, IGUCR, and ERPF were still independently associated with DR.
Conclusion: In 2-DM patients, tubular indexes and DR were the sensitive and noninvasive biomarkers for early prediction of diabetic kidney disease.

Disclosure
L. Zhang: None. J. Lu: None.
526-P: Urine Tricarboxylic Acid Cycle Organic Anions and Progression of Diabetic Kidney Disease
Authors: LUNYERA JOSEPH , NGUYEN MAGGIE , DIAMANTIDIS CLARISSA J. , BOSWORTH HAYDEN B. , PATEL UPTAL D. , DAVENPORT CLEMONTINA A. , BAIN JAMES R. , MUEHLBAUER MICHAEL , ILKAYEVA OLGA , SHAH SVATI H. , SCIALLA JULIA 
Published: June 2020

Background: Urine tricarboxylic acid (TCA) cycle organic anions (OAs) are elevated in diabetes and may be biomarkers for diabetic kidney disease (DKD) progression.
Methods: We conducted an ancillary study to the Simultaneous Risk Factor Control Using Telehealth to Slow Progression of Diabetic Kidney Disease (STOP-DKD) Trial - a randomized trial of pharmacist-led medication and behavior management in 281 patients with early to moderate DKD at Duke from 2014-2015. In this study, we used linear mixed models to assess associations of 10 urine TCA cycle OAs with estimated glomerular filtration rate (eGFR) and eGFR decline. TCA cycle OAs were measured in 24h urine samples by targeted MS and modeled as: (1) the average of z-scores for each OAs; and (2) factor scores derived by principal component analysis (PCA). We extended follow-up for eGFR to mid-2019 using electronic health records.
Results: Among 132 participants (50% men; 58% Black; mean age 64 years [SD 9]; median eGFR 72 ml/min/1.73m2 [IQR 59-89] and urine albumin-to-creatinine [UACR] 25 mg/g [IQR 9-81]), PCA identified 3 OA metabolite factors (F). Fumarate, malate, α-ketoglutarate, citrate, lactate, and pyruvate loaded positively on F1; succinate, pyruvate, and lactate loaded negatively on F2; and methylsuccinate, ethylmalonate, and methylmalonate loaded positively on F3. Over a median follow-up of 1 year [IQR 1.3], higher average OA z score, F1, and F3 were each strongly associated with higher baseline eGFR after adjustment for age, sex, race, and log(UACR) at baseline (all p&lt;0.01). Higher F2, but not other scores, was associated with faster eGFR decline (p=0.05).
Conclusion: Lower urine TCA cycle OAs, most notably a factor comprising lower succinate, pyruvate and lactate, are potential biomarkers for faster DKD progression.

Disclosure
J. Lunyera: None. M. Nguyen: None. C.J. Diamantidis: None. H.B. Bosworth: Board Member; Self; Preventric Diagnostic. Consultant; Self; Abbott, Medicines Company, Novartis Pharmaceuticals Corporation. Research Support; Self; improved patient outcomes, Novo Nordisk Foundation, Otsuka Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., Pharma foundation, Proteus Digital Health, Proteus Digital Health, Sanofi. U.D. Patel: Board Member; Self; Goldfinch Bio. Employee; Self; Gilead Sciences, Inc. C.A. Davenport: None. J.R. Bain: None. M. Muehlbauer: None. O. Ilkayeva: None. S.H. Shah: Research Support; Self; AstraZeneca, Verily Life Sciences LLC. J. Scialla: Advisory Panel; Self; Tricida. Other Relationship; Self; GlaxoSmithKline plc., Sanofi.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (DK076169)