bluuebunny/crossref_metadata_2025 huggingface dataset:
Gut Incretin Release as a Mediator of Metabolic Effects of Metformin in Type 2 Diabetic Patients with Chronic Heart Failure
Authors: STOLARIKOVA EVA , KOPECKY JAN , VELEBA JIRI , VELEBOVA KATERINA , BELINOVA LENKA , MELENOVSKY VOJTECH , KOPECKY JAN , PELIKANOVA TEREZIE
Published: June 2018

Introduction: Metformin (MET) is a 1st choice drug used in patients with type 2 diabetes (DM). Treatment with MET in patient with DM and heart failure (HF) is associated with better cardiovascular outcomes, but mechanisms of MET-mediated effects are unexplained. Our aim was to evaluate the effects of MET on substrate metabolism, cardiac function and structure in patient with DM and HF.

Methods: 40 patients with DM and stable chronic HF (age: 59 ± 9 years) were studied in a random sequence cross-over clinical study testing the effect of 3-month usage of MET vs. placebo. The subjects were randomized to MET (2 g/day) or to placebo group. After the three months of treatment the medication was changed and the treatment continued for next 3 months. At the beginning and the end of each intervention period (3 times in total) the panel of various metabolic and cardiovascular tests was done: echocardiography, spiroergometry and meal test with evaluation of several variables like incretins (GLP-1 and peptide YY) and parameters of oxidative stress (MCP-1, TNFα).

Results: Compared to placebo, MET was accompanied with significantly higher increase of selected gut-related hormones. GLP-1 area-under-curve increased during the meal test (MET vs. PL: 8571.39±6899.57 vs. 5665.82±4580.57pmol.hour.l-1; p<0.01). Also the peptide YY area-under-curve increased during the meal test (MET vs. PL: 29809.67±13442.43 vs. 23636.21±8183.08pmol.hour.l-1; p<0.05). MCP-1 showed inconsistent trend to increase during meal test. Fasted levels of TNFα were higher after MET, but there was no change during the meal test compared to placebo. MET therapy led to reduction of HbA1C (p<0.001) and reduced glucose area-under-curve (p<0.001) during glucose tolerance test. MET had neutral effect on cardiac structure and function.

Conclusions: Our results suggest that metabolic effects of MET might be mediated by improvement of gut endocrine function. MET had neutral effect on cardiac function or structure.

Disclosure

E. Stolarikova: None. J. Kopecky: None. J. Veleba: None. K. Velebova: None. L. Belinova: None. V. Melenovsky: None. J. Kopecky: None. T. Pelikanova: None.
Novel "Dual Hit" Rat Model of Diabetic Cardiomyopathy
Authors: THISTED LOUISE , LINDSAY ROSS T. , FOSGERAU KELD , SECHER THOMAS , THOMSEN MORTEN B. , JESPERSEN THOMAS , MURRAY ANDREW J. , PEDERSEN PHILIP J. , VRANG NIELS , FINK LISBETH N. , PEDERSEN TANJA X. , ZOIS NORA E.
Published: June 2018

The pathogenesis of diabetic cardiomyopathy (DC) is poorly understood and new drugs targeting the myocardium are absent. This may be ascribed to the failure of available pre-clinical models to recapitulate essential clinical features of DC and heart failure. We hypothesized that the combination of experimental diabetes and pharmacologically induced cardiac stress might provide a novel rat model displaying a distinct profile of DC. Non-insulin dependent type 1 like diabetes mellitus was induced by partial (90%) pancreatectomy (Px). Pronounced hyperglycemia was established within two weeks (blood glucose levels of 23.3 ± 3.7 mM). Five weeks post Px or sham surgery, vehicle or the sympathomimetic agent, isoproterenol (Iso, 1 mg/kg, SC) was administered for 10 days. Ten weeks after surgery, the heart was isolated for detailed histological and molecular characterization. The relative weight of the left ventricle (LV) was significantly increased in Px-Iso rats compared to sham-vehicle treated control rats (0.22 ± 0.03 vs. 0.25 ± 0.02 mg/g BW, p<0.05), indicating Px-Iso-induced cardiac hypertrophy. Compared to sham-vehicle rats, cardiac muscle fibers of Px-vehicle rats had attenuated respiratory function and increased reliance upon oxidation of fatty acid substrates, measured by high-resolution respirometry. Interestingly, this aggravation was reversed in Px-Iso rats. Moreover, Iso induced cardiac fibrosis demonstrated by quantitative histology (area fraction; Px-vehicle vs. Px-Iso: 3.1 ± 0.2% vs. 6.3 ± 0.5%; Sham-vehicle vs. Sham-Iso: 3.7 ± 0.4% vs. 7.6 ± 0.5%, p<0.001). Echo- and electrocardiography as well as transcriptome analyses were applied to further assess LV remodeling and cardiac function.

In conclusion, the Px-Iso rat model may provide a state-of-the-art model of DC displaying key features of the clinical condition. Hereby, this model may be useful in the evaluation of cardiovascular effects of novel compounds in the pre-clinical phase of drug development.

Disclosure

L. Thisted: Employee; Self; Gubra. R.T. Lindsay: None. K. Fosgerau: Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Self; Gubra. Stock/Shareholder; Self; Gubra. T. Secher: Employee; Self; Gubra. Employee; Spouse/Partner; Novo Nordisk A/S. M.B. Thomsen: None. T. Jespersen: None. A.J. Murray: None. P.J. Pedersen: None. N. Vrang: Board Member; Self; Gubra. Employee; Self; Gubra. Stock/Shareholder; Self; Gubra. L.N. Fink: Employee; Self; Gubra. Stock/Shareholder; Self; Novo Nordisk A/S. T.X. Pedersen: Employee; Self; Gubra. N.E. Zois: Employee; Self; Gubra.
Effects of NAFLD on Acetyl-CoA Partitioning and Ketone Kinetics in Response to a 24-Hour Fast
Authors: FLETCHER JUSTIN A. , DEJA STANISLAW , BURGESS SHAWN C. , BROWNING JEFFREY
Published: June 2018

During an overnight fast (12 hour), the liver provides glucose as fuel for other tissues via glycogenolysis and the energetically expensive process of gluconeogenesis. During this period, energy to support this process is generated primarily by the tricarboxylic acid (TCA) cycle. As the duration of fasting prolongs, and glycogen stores are depleted (>24 hour), the liver shifts to the production of ketone bodies that can be used as a secondary fuel source by peripheral tissues. In healthy humans, this period of fasting is generally associated with increased β-oxidation and partitioning of acetyl-CoA away from the TCA cycle and toward ketone body production. We have previously shown that subjects with nonalcoholic fatty liver disease (NAFLD) have increased rates of acetyl-CoA oxidation in the TCA cycle after a 12 hour fast. To examine how NAFLD affects hepatic response to a 24 hour fast, we infused stable isotope tracers into BMI and age matched subjects with (n=12) and without (n=12) NAFLD. At enrollment, NAFLD subjects had greater liver triglycerides (P<0.05) and hepatic insulin resistance (P<0.05), as well as higher fasting plasma concentrations of cholesterol, triglycerides, and glucose (P<0.05). After fasting for 24 hours, subjects with NAFLD failed to suppress endogenous glucose production as compared to controls. This was associated with a higher ratio of plasma insulin to glucagon as well as a blunted increase in the concentration of plasma free fatty acids as fasting duration prolonged. As previously shown in overnight fasted subjects, TCA cycle activity was significantly higher in NAFLD subjects after a 24 hour fast. However, despite elevated TCA cycle flux, NAFLD subjects failed to increase overall β-oxidation compared to controls due to reduced utilization of acetyl-CoA for ketone synthesis.

In conclusion, after a 24 hour fast, the liver of NAFLD subjects produced more glucose and disposed of less fat via β-oxidization by shuttling more acetyl-CoA to the TCA cycle but much less to ketogenesis.

Disclosure

J.A. Fletcher: None. S. Deja: None. S.C. Burgess: None. J. Browning: None.
Nrf2 Exaggerates Cardiomyopathy Associated with Type 1 Diabetes in Mice
Authors: ZANG HUI M. , WU WEIWEI , QI LEI , BOWEN CHRISTOPHER , CUI TAIXING , NAGARKATTI PRAKASH , NAGARKATTI MITZI
Published: June 2018

Nuclear factor-erythroid factor 2-related factor 2 (Nrf2), a transcription factor for cellular defense against oxidative stress and other insults, has been postulated to be an inhibitor of diabetes and diabetic cardiomyopathy. However, this notion has been challenged by the detrimental role of Nrf2 in the pathogenesis of diabetes and diabetic cardiomyopathy. Thus, we investigated the nature of the Nrf2-mediated dichotomy in a mouse model of streptozotocin (STZ)-induced type 1 diabetes over a time period of 9 months. We found that global Nrf2 knockout (Nrf2 KO) increased the death rate in the setting of STZ-induced diabetes, but it partly protected against STZ-induced diabetes and insulin-resistance in both male and female mice. Nrf2 KO hardly affected the onset and the progression of cardiac dysfunction associated with the diabetes by 6 months in both male and female mice; however, it partly prevented the progression of cardiac dysfunction at 9 months. Cardiomyocyte hypertrophy, cardiac fibrosis, and cardiac oxidative stress associated with the diabetes not at 3 months but at 6 and 9 months were inhibited by Nrf2 KO in both male and female mice. In addition, cardiomyocyte-restricted (CR) Atg5 KO had minimal impact on the blood glucose level, but exaggerated the diabetic cardiomyopathy, which could be attenuated in part by additional Nrf2 KO. At the molecular level, diabetes-induced autophagy impairment turned off Nrf2-operated antioxidant defense while switching on Nrf2-mediated detrimental signaling to cardiomyocytes. Taken together, our findings reveal that Nrf2 is cardiac protective when myocardial autophagy is intact; however, it becomes detrimental to the heart when myocardial autophagy is impaired due to a chronic diabetic setting.

Disclosure

H.M. Zang: None. W. Wu: None. L. Qi: None. C. Bowen: None. T. Cui: None. P. Nagarkatti: None. M. Nagarkatti: None.
Poldip2 Affects Collagen I Accumulation by Regulating the Expression of Zyxin in Vascular Smooth Muscle Cells
Authors: FUJII MASAKAZU , SONODA NORIYUKI , OKAMOTO MISATO , MORINAGA HIDETAKA , OGAWA YOSHIHIRO , GRIENDLING KATHY
Published: June 2018

Objectives: Polymerase delta interacting protein 2 (Poldip2) has previously been implicated in migration, proliferation and extracellular matrix (ECM) production in vascular smooth muscle cells. Thereby, regulation of Poldip2 protein expression is suggested to prevent the development and progression of diabetic macroangiopathy. We recently reported that Poldip2+/- mouse aortic smooth muscle cells (MASMs) exhibit higher β1-Integrin expression and activity of the PI3K/Akt/mTOR signaling pathway, leading to increased ECM protein synthesis. We further investigated the mechanism of Podlip2 in regulating the β1-Integrin expression.

Approach and Results: Cell adhesion and cell signaling, below the focal adhesion, were investigated. Poldip2+/- MASMs (HET) showed both quicker cell attachment and detachment than Poldip2+/+ MASMs (WT). Paxillin, one of the components of focal adhesion, is expressed in all areas (from front to back) of lamellipodia in WT, on the other hand HET express the Paxillin only front area, which may induce weaker cell adhesion. The mature focal adhesion marker, Zyxin, expression is decreased in HET significantly. Intriguingly, the activity of RhoA is increased in HET and induced MRTF-A translocation from cytosol to nacreous.

Conclusions: It is revealed that Poldip2 is positive regulator of Zyxin. HET has weak cell adhesion, and may induce Poldip2+/- MASMs exhibit higher β1-Integrin expression as a compensation against immature focal adhesion. Thus PI3K/Akt/mTOR signaling pathway is activated, leading to increased ECM protein synthesis. Translocation of MRTF-A from cytosol to the nucleus, may contribute to bind to transcriptional factor for Collagen I. This phenomenon is suggested as one more pathway, which Collagen I accumulates in HET. These findings have important implications for atherosclerotic vascular diseases such as diabetic macroangiopathy in which ECM accumulation plays a role.

Disclosure

M. Fujii: None. N. Sonoda: None. M. Okamoto: None. H. Morinaga: None. Y. Ogawa: Research Support; Self; AstraZeneca. Advisory Panel; Self; Gilead Sciences, Inc.. Research Support; Self; Japan Agency for Medical Research and Development, Japan Society for the Promotion of Science, Mitsubishi Tanabe Pharma Corporation, Mochida Pharmaceutical Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd.. K. Griendling: None.
Renin-Angiotensin System Inhibition Alters Triacylglycerol Metabolism in Diabetic Kidney Disease
Authors: SAS KELLI , LIN JIAHE , NAIR VIJI , KRETZLER MATTHIAS , BROSIUS FRANK , PENNATHUR SUBRAMANIAM
Published: June 2018

Lipids are essential metabolites with diverse functions impacting physiology. We recently identified several lipidomic abnormalities in diabetic kidney disease (DKD), including remodeling of triacylglycerol (TAG) fatty acid composition to increase availability of long, polyunsaturated TAGs. Using the BKS eNOS-/- db/db diabetic mouse model, we studied the effect of renin-angiotensin system (RAS) inhibition with combination treatment of lisinopril (20 mg/kg/d) and losartan (30 mg/kg/d) for 12 weeks, an intervention which ameliorates DKD in this mouse model without impacting glycemia. We examined the kidney cortical lipidome to identify lipid alterations that may be pathogenic in DKD. As expected, RAS inhibition did not significantly alter glycemic control or total plasma triglycerides or cholesterol, but decreased the 24-hour urine albumin/creatinine ratio (2564 vs. 615 μg/mg creatinine) and decreased the glomerular periodic acid-Schiff stained area by 13% (p<0.05). We performed mass-spectrometry based untargeted lipidomic profiling to quantify 635 lipids across 17 lipid classes in kidney cortex. Lisinopril/losartan treatment remodeled TAG fatty acid composition and increased conversion between TAGs and several glycerophospholipid classes, particularly medium-chain phosphatidylcholines, phosphatidylethanolamines, phosphatidylinositols and cardiolipins (p<0.05). These correlations in lipid levels were associated with changes in corresponding enzymes of lipid metabolism, suggesting increased metabolism of TAGs into glycerophospholipids, which are key components of cell membranes and precursors for signaling molecules. In addition to the known salutary effect of RAS inhibition on DKD, our results suggest a previously unrecognized role of RAS inhibition on kidney complex lipid levels through increased TAG metabolism. These findings raise the possibility that TAG metabolism might be a target for intervention aimed at decreasing DKD progression.

Disclosure

K. Sas: None. J. Lin: None. V. Nair: None. M. Kretzler: Research Support; Self; AstraZeneca, Novo Nordisk Inc., Boehringer Ingelheim GmbH, National Institute of Diabetes and Digestive and Kidney Diseases, Eli Lilly and Company, Gilead Sciences, Inc.. F. Brosius: None. S. Pennathur: None.
Histone H3 Serine 10 Phosphorylation Facilitates Endothelial Activation in Diabetic Kidney Disease
Authors: ALGHAMDI TAMADHER , BATCHU SRI NAGARJUN , HADDEN MITCHELL J. , LIU YOUAN , BOWSKILL BRIDGIT , GELDENHUYS LAURETTE , SIDDIQI FERHAN S. , MAJUMDER SYAMANTAK , ADVANI ANDREW
Published: June 2018

Post-translational modification of histone proteins epigenetically affects gene transcription. The histone modifications that have been most extensively linked to the development of diabetes complications are histone (de)methylation and (de)acetylation, whereas other modifications have been comparatively overlooked. Here, by examining the means by which podocytes may influence the glomerular endothelial phenotype, we identified a role for the phosphorylation of histone H3 on serine residue 10 (H3Ser10) in mediating endothelial activation in diabetes. Culture media conditioned by high glucose-exposed podocytes induced glomerular endothelial vascular cell adhesion molecule-1 (VCAM-1) upregulation and was enriched for the chemokine CCL2 in a JAK2-dependent manner. Antagonism of CCR2, the cognate receptor for CCL2, decreased the endothelial VCAM-1 upregulation that was induced either by conditioned medium or by recombinant CCL2, whereas CCR2 knockout mice exhibited diminished glomerular VCAM-1 upregulation when challenged by diabetes. Recombinant CCL2 induced glomerular endothelial VCAM-1 upregulation through a pathway mediated by p38 MAPK, its nuclear substrates MSK1/2 and subsequent H3Ser10 phosphorylation, which was enriched at the VCAM-1 promoter. H3Ser10 phosphorylation levels were elevated in the kidneys of streptozotocin-diabetic endothelial nitric oxide synthase knockout (STZ-eNOS-/-) mice and in the glomeruli of humans with diabetic kidney disease. Finally, treatment of STZ-eNOS-/- mice with an inhibitor of JAK2 decreased both urinary CCL2 excretion and renal H3Ser10 phosphorylation. Collectively, these findings indicate that histone phosphorylation promotes endothelial activation in diabetes. Histone protein phosphorylation should be set alongside other more canonical histone modifications when considering chromatin modifications that influence diabetic kidney disease and that may be amenable to therapeutic intervention.

Disclosure

T. Alghamdi: None. S. Batchu: None. M.J. Hadden: None. Y. Liu: None. B. Bowskill: None. L. Geldenhuys: None. F.S. Siddiqi: Research Support; Self; AstraZeneca. S. Majumder: None. A. Advani: Research Support; Self; AstraZeneca, Boehringer Ingelheim GmbH. Other Relationship; Self; Boehringer Ingelheim GmbH.
Transplantation of P53-Silenced Endothelial Progenitor Cells (EPCs) Under Renal Capsule—A Potential Treatment for Diabetic Kidney Disease (DKD)
Authors: KUNDU NABANITA , ASICO LAUREANO D. , DOMINGUES CLEYTON C. , AHMADI NEEKI , SEN SABYASACHI
Published: June 2018

Diabetic kidney disease (DKD) is one of the major vascular complications of diabetes, which is associated with glomerulosclerosis and poor perfusion. Therefore, improving the renal perfusion may help to halt or reverse the kidney injury. Here, we investigated whether transplanting mouse EPC (that has been modified by transiently silencing p53 gene using Adenovirus transduction ex-vivo), under renal capsule could improve angiogenesis and renal perfusion, by preventing EPC apoptosis in a hyperglycemic milieu.

Methods: First, we confirmed hyperglycemia and progressive proteinuria in STZ-induced type 1 diabetic C57BL/6J mouse. Next, we transplanted 0.3 million p53-silenced EPCs,or Ad-Null-EPCs (control), bilaterally, under each kidney capsule. Another comparator was non-STZ normal mouse. Urine was collected weekly for volume and protein estimation. Renal blood flow was measured by laser Doppler. Kidneys were harvested post sacrifice and qRT-PCR were performed for targeted (towards angiogenic genes) gene expression assays.

Results: Excess urine volume as noted in the hyperglycemic mouse models was reduced 4-fold post transplantation of p53sh-EPCs compared to null. There was no proteinuria after week 2 in p53sh-EPC transplanted mouse unlike control. Enhanced blood flow by laser doppler (3.2 fold) was also noted with delivery of p53sh-EPCs compared to null EPC. Absence of proteinuria (after 2 weeks) and renal blood flow measurements for p53sh group were similar to the non-STZ mouse. Interestingly, markers for neovascularization, such as eNOS (4.5 fold, p=0.002) and VEGF-A (1.5 fold, p=0.03) upregulated significantly post p53 silenced EPC transplantation compared to null EPC. CD31 staining is pending.

Conclusion: Transient silencing of p53 gene in mouse EPCs help to improve proteinuria, diabetic polyuria and renal blood flow, most likely by increasing angiogenesis and perfusion and may have a prominent therapeutic role in DKD.

Disclosure

N. Kundu: None. L.D. Asico: None. C.C. Domingues: None. N. Ahmadi: None. S. Sen: None.
Renal Mitochondrial Oxidative Stress Induced by NAD+-Dependent Sirt3 Inactivation via Overexpression of CD38 (NAD+ase) in Diabetic Kidney Disease
Authors: OGURA YOSHIO , KITADA MUNEHIRO , MONNO ITARU , WATANABE AI , KOYA DAISUKE
Published: June 2018

Diabetic kidney disease (DKD) is a major cause of end-stage renal disease worldwide. Although the detailed pathogenesis of DKD is not elucidated yet, aging is recognized as one of the risk factors for the development of end-stage renal failure due to chronic kidney disease including DKD. Therefore, investigation of aging-related mechanism would be a novel therapeutic target for DKD. NAD levels decrease during aging. Previous report demonstrated that expression and activity of the CD38 (NADase) increased with aging, and that CD38 is required for the age-related NAD decline and mitochondrial dysfunction via a pathway mediated at least in part by regulation of Sirt3 activity. However, the role of CD38 in the pathogenesis for DKD has never been investigated. In this study, we evaluated the role of CD38 on the mechanisms of renal mitochondrial oxidative stress which is related to changing Sirt3 activity, in Zucker Diabetic Fatty rats (ZDFRs) and in cultured HK2 cells under high-glucose condition. At 28 weeks of age, the ZDFRs exhibited elevated HbA1c levels, heavier kidney weight, increased urinary albumin, L-FABP and 8-OHdG excretion, histological tubulo-interstitial fibrosis and tubular cell damage, compared to nondiabetic Zucker Lean rats. Additionally, in renal mitochondria, the NAD+/NADH ratio was reduced, and acetylation levels of mitochondrial antioxidant enzymes, IDH2 and SOD2, which are regulated by Sirt3, were increased in ZDFRs. Similarly, in cultured HK2 cells under high-glucose condition, CD38 expression was increased, resulting in the reduction of NAD+/NADH ratio and Sirt3 activity, which increased in acetylation levels of IDH2 and SOD2. Administration of the CD38 inhibitor, apigenin, to ZDFRs and HK2 cells, restored the NAD+/NADH ratio, decreased the levels of IDH2 and SOD2 acetylation. Therefore, restoring Sirt3 activation by suppression of CD38 may be a novel therapeutic target for DKD.

Disclosure

Y. Ogura: None. M. Kitada: Research Support; Self; Astellas Pharma US, Inc., Kyowa Hakko Kirin Co., Ltd., Daiichi Sankyo Company, Limited, Ono Pharmaceutical Co., Ltd., Novartis Pharma K.K., Sumitomo Dainippon Pharma Co., Ltd., Boehringer Ingelheim GmbH, Taisho Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Corporation. I. Monno: None. A. Watanabe: None. D. Koya: None.
Novel Rat Models of Diabetic Nephropathy Displaying Pronounced Tubular Fibrosis and Glomerular Alterations
Authors: SECHER THOMAS , JOHANSEN THEA T. , OESTERGAARD METTE V. , PEDERSEN PHILIP J. , ZOIS NORA E. , PEDERSEN TANJA X. , VRANG NIELS , FOSGERAU KELD , FINK LISBETH N.
Published: June 2018

Diabetic nephropathy (DN) is a long-term complication of diabetes characterized by proteinuria and loss of kidney function. Up to one third of patients with diabetes develop DN, and no new therapies targeting DN have been introduced for more than a decade. The lack of treatment options may, in part, be caused by the scarcity of translatable rodent models. We have established a surgically induced rat model of non-insulin dependent type 1-like diabetes (90% pancreatectomy, Px), displaying a stable blood glucose of >20 mmol/L. In addition, both Px and uninephrectomy (UNx) was performed simultaneously to potentially increase progression of renal pathological changes. Kidneys in the Px diabetic rat model displayed hypertrophy of glomeruli, cortex, and medulla 11 weeks post-surgery. Renal fibrosis was significantly increased in Px rats vs. nondiabetic control animals, as determined by quantitative histology following picro-sirius red staining of collagen (total kidney collagen, Px vs. controls; 127 ± 8 mg vs. 39 ± 5 mg, p<0.001). Plasma urea was increased in Px rats compared to nondiabetic animals (8.3 ± 0.6 vs. 5.5 ± 0.3 mmol/L, p<0.01). Finally, gene expression of the kidney injury markers NGAL and KIM1 as well as gene expression of pro-fibrotic macrophage markers (mannose receptor, MGL1, and CD163) was increased in Px rat kidneys. Px-UNx rats developed a diabetic phenotype similar to that observed in Px rats. In contrast, a highly increased kidney weight was detected with the more advanced surgical approach. Detailed histological analyses will establish whether glomerular and tubular pathology is exacerbated to a comparable extent.

In conclusion, our data suggest that the Px model develops profound renal hypertrophy and fibrosis, and that renal hypertrophy is exacerbated in the Px-UNx model. Thus, Px alone and Px-UNx in rats may represent novel, strong alternatives to streptozotocin-induced diabetes and genetic models for the study of DN drug candidates.

Disclosure

T. Secher: Employee; Self; Gubra. Employee; Spouse/Partner; Novo Nordisk A/S. T.T. Johansen: None. M.V. Oestergaard: None. P.J. Pedersen: None. N.E. Zois: Employee; Self; Gubra. T.X. Pedersen: Employee; Self; Gubra. N. Vrang: Board Member; Self; Gubra. Employee; Self; Gubra. Stock/Shareholder; Self; Gubra. K. Fosgerau: Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Self; Gubra. Stock/Shareholder; Self; Gubra. L.N. Fink: Employee; Self; Gubra. Stock/Shareholder; Self; Novo Nordisk A/S.
Early Growth Response Protein 1 Mediates Long Noncoding RNA Arid2-IR to Promote Extracellular Matrix Production in Diabetic Kidney Disease
Authors: YANG YAN-LIN , HU FANG , XUE MENG , JIA YIJIE , ZOU MEINA , XUE YAO-MING
Published: June 2018

Background: In order to investigate the mechanism through which Early growth response protein 1 (Egr1) mediates long noncoding RNA Arid2-IR to promote diabetic kidney disease.

Method: In vivo, we induced type 2 diabetes and overexpressed or knocked down Egr1 by rapidly injecting of a large volume of DNA solution through the tail vein. In vitro, we assessed mouse mesangial cells that were stimulated with high glucose conditions and transiently transfected with siRNA and plasmids.

Results: Compared with those in the control group, the expression of Egr1 and Arid2-IR in diabetes mice increased with the up-regulation of Col1a1 and α-SMA. Arid2-IR expression changes with Egr1 knockdown and overexpression both in vivo and in vitro. Knockdown Arid2-IR in vitro reduced Col1a1 and α-SMA expression. Overexpressing Egr1 and silencing Arid2-IR reduced Col1a1 and α-SMA expression.

Conclusion: Egr1 promoted ECM production by up-regulating Arid2-IR. Thus, Arid2-IR may be a new target for treating diabetic kidney disease.

Disclosure

Y. Yang: None. F. Hu: None. M. Xue: None. Y. Jia: None. M. Zou: None. Y. Xue: None.
miR-877-3p Induces Apoptosis by Targeting BCL2 in Tubular Epithelial Cells
Authors: ZOU MEINA , XUE MENG , HU FANG , JIA YIJIE , YANG YAN-LIN , XUE YAO-MING
Published: June 2018

Background: Apoptosis of renal tubular epithelial cells plays an important role in the progression of diabetic kidney disease(DKD). Our previous study found that miR-877-3p of urinary exosomes significantly increased in patients with DKD, but its mechanism of action in DKD remains unclear. Here, we found that miR-877-3p significantly aggravated the progression of diabetic kidney disease. miR-877-3p was found to significantly increase tubular epithelial cell apoptosis and appeared to act by down-regulating the anti-apoptotic gene BCL2 in human proximal tubule cell line(HK-2 cell). Bioinformatics software and luciferase reporter gene analysis showed that miR-877-3p promotes apoptosis and aggravates the progression of DKD through targeting BCL2 3’UTR region in HK-2 cells.

Methods: miR-877-3p and BCL2 were detected in HFD/STZ-induced mice upon their sacrifice at 6 and 12 w. In vitro studies, we incubated HK-2 cells under high-glucose (HG) to mimic DKD.Then,BCL2 expression was detected in HK-2 cells with overexpression of miR-877-3p. BCL2, Bax and cleaved caspase3 gene and protein expression levels were determined by RT-qPCR and western blotting, respectively.The direct relationship between miR-877-3p and BCL2 was explored via luciferase reporter assay.

Results: miR-877-3p expression was more significantly increased in the renal cortices of diabetes mellitus (DM) mice at 12 w after treatment compared to the renal cortices of 6W.In contrast, BCL2 expression was significantly decreased in DM mice compared to control mice only at 12 w after treatment.We also found that miR-877-3p was increased in HG treated HK-2 cells.Overexpression of miR-877-3p in HK-2 cells downregulate the expression of BCL2.

Conclusions: miR-877-3p promotes DKD progression through targeting BCL2.

Disclosure

M. Zou: None. M. Xue: None. F. Hu: None. Y. Jia: None. Y. Yang: None. Y. Xue: None.
Improved Diabetes Control Using SMBG Pattern Management in High-Risk Minorities (HRM)
Authors: MYERS IKENNA
Published: June 2018

SMBG in a diabetes education program improves T2D outcomes. This study was done to evaluate if use of SMBG taught by clinical pharmacists or Certified diabetes educators improved A1C compared to diabetes classes and medication review in HRM. A pretest-posttest format was used. Subjects (n=807) were referred by providers from 5 federally qualified health centers and a practice group.

HEDIS measures collected included: A1C, BP, FPG, BMI, total cholesterol (TC), LDL-C, HDL-C, and TGs. Subjects received a validated knowledge test initially, 3 months after taking a diabetes class and at study end. A psychosocial inventory was collected initially and at study end. This multi-component model was a four-arm study comparing Diabetes Classes (DC), Pharmacist Medication Management (MTM), Pattern Management (PM) using a glucose meter data management system, and control subjects who did not participate (Ctl). Subjects had at least two interventions prior to assignment to PM. 258 subjects attended the diabetes classes, 71 in the MTM arm, 44 in the PM arm, and 138 Ctls. 296 were lost due to incomplete and/or lost data. Demographics revealed 77% in the 20-64 years old range, 21% in the >65 age group, and 16% in the <20 age group, with 65% female subjects. The mix was 86% non-Hispanic black and 13% non-Hispanic white. 60% of subjects were on Medicare/Medicaid, 30% employer-insured, and 16% uninsured. Mean A1C levels in the DC group decreased from 7.93 to 7.60, 8.06 to 7.32 in the MTM group, 7.70 to 7.28 in the PM group, while Ctls increased from 7.00 to 7.36. The PM group had the greatest mean changes in multiple measures including BMI, BP, FP, TC, and TG. The MTM group had the greatest improvement in A1C.

Conclusions: The PM group had the largest changes in overall measures of T2D control. There was increased diabetes knowledge and reduced ER visits. Thus, in HRM using PM from SMBG, the improvements suggested better monitoring of multiple metrics of relevance in T2D control.

Disclosure

I. Myers: None.
TRIM29 Is a New Gene That Regulates IRS-1 to Induce Insulin Resistant in Diabetes
Authors: HABIB SAMY L.
Published: June 2018

Insulin resistance is a major risk for development of type 2 diabetes. Several genes involved in regulation of insulin receptor substrate 1 (IRS1) and lead to chronic diabetes. Our RNA sequence data from kidney of diabetic mice show a new candidate gene involved in the regulation of IRS1. New candidate gene, Tripartite motif protein 29 (TRIM29), acts as an E3 ligase targeting insulin receptor (IR) and insulin receptor substrate 1 (IRS1) for ubiquitin-dependent degradation, resulting in insulin resistance. RNA seq data show that TRIM29 is higher 2.5 and 3.6 fold in kidney cortex of diabetic mice (db/db) at age of 10 and 12 months compared to wild type mice, respectively. Mouse renal proximal tubular epithelial cells treated with high glucose (25mM) for 48 hours showed significant increased in TRIM29 and decreased in IRS-1 compared to cells grown in normal glucose (5mM). Pre-transfected cells with siRNA against TRIM29 resulted in significant increased IRS-1 and cell proliferative marker, cyclin D1 expression. Downregulation of tuberin by siRNA significantly decreased TRIM29 expression and increased cyclin D1 expression. In addition, downregulation of AMPK by DN-AMPK and compound C resulted in significant decreased TRIM29 compared to cells transfected with control-siRNA. Kidney cortex of db/db mice and WT mice at age 10 and 12 months were analyzed by Western blot showed that TRIM29 is significantly increased and IRS-1 decreased compared to wild type mice. Increase TRIM29 in kidney diabetic mice was associated with increased in cyclin D1. Here, we identified a new candidate gene, TRIM29 that involves in the regulation IRS-1 pathways to increase cell proliferation and enhance insulin resistance diabetes. The new gene, TRIM29, may be used as a new candidate target to treat insulin resistant in diabetes.

Disclosure

S.L. Habib: None.
Circulating MicroRNAs Associated with Incident End-Stage Renal Disease in Chinese with Type 2 Diabetes
Authors: FAN BAOQI , LEE HEUNG MAN , LIM CADMON K.P. , CHOY RICHARD , CHAN JULIANA C. , LUK ANDREA , MA RONALD C.
Published: June 2018

Objectives: We aimed to identify novel serum micro(mi)RNAs associated with incident end-stage renal disease (ESRD) in Chinese with type 2 diabetes.

Methods: We conducted a nested case-control study in patients with type 2 diabetes enrolled in the Hong Kong Diabetes Registry between 1995 and 2007. Cases were patients who were free from ESRD at enrolment but developed renal endpoint as defined as eGFR < 15 ml/min/1.73m2 or dialysis during follow-up until 2015. Controls were patients who had normal renal function at both baseline and during observation period. MicroRNAs were extracted from stored serum collected at baseline. Discovery cohort included 22 cases and 21 controls and miRNAs were screened by Agilent microRNA microarray. Validation cohort included 361 ESRD cases and 241 T2D controls and miRNAs were quantitated using qRT-PCR. Two spike-in miRNAs were used to control for efficiency in RNA extraction and reverse transcription. Batch difference was adjusted by a positive RNA control.

Results: In the discovery analysis, miR-X and miR-Y were elevated with respective fold-change of 5.97 and 4.43 in patients with incident ESRD compared to those without renal events on follow-up. Pathway analysis revealed that the two miRNAs may involve in signaling transduction, adrenergic signaling, or relate to peroxisome. Increased levels of miR-X and miR-Y were also detected in the validation cohort. Using binary logistic regression adjusted for age, sex and disease duration, doubling of miR-X and miR-Y were associated with 14.2% and 26.3% higher odds of progressing to ESRD, respectively. The association remained significant for miR-Y when further adjusted for metabolic indices, baseline albuminuria and eGFR.

Conclusions: Baseline serum levels of miR-X and miR-Y are independently associated with higher risk of incident ESRD. These results indicate the potential of circulating miRNAs to serve as prognostic indicators for disease progression.

Disclosure

B. Fan: None. H. Lee: None. C.K.P. Lim: None. R. Choy: None. J.C. Chan: Consultant; Self; Bayer AG. Other Relationship; Self; Bayer AG. Consultant; Self; Sanofi. Other Relationship; Self; Sanofi, Eli Lilly and Company, Amgen Inc.. Consultant; Self; AstraZeneca, Merck & Co., Inc., Pfizer Inc.. Other Relationship; Self; Pfizer Inc.. Board Member; Self; Asia Diabetes Foundation. Stock/Shareholder; Self; GemVCare. Other Relationship; Self; Merck Sharp & Dohme Corp.. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Novartis AG, Eli Lilly and Company. A. Luk: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc.. Research Support; Self; Sanofi. R.C. Ma: Research Support; Self; AstraZeneca, Bayer AG, Merck Sharp & Dohme Corp., Pfizer Inc.. Advisory Panel; Self; Boehringer Ingelheim GmbH, Nippon Boehringer Ingelheim Co. Ltd..
Empagliflozin Improves Renal Outcomes Irrespective of Control of Blood Pressure, Low-Density Lipoprotein Cholesterol, and HbA1c
Authors: WANNER CHRISTOPH , COOPER MARK E. , INZUCCHI SILVIO , ZINMAN BERNARD , HEHNKE UWE , EYNATTEN MAXIMILIAN VON , KOITKA-WEBER AUDREY
Published: June 2018

The EMPA-REG OUTCOME trial demonstrated the renoprotective role of the sodium-glucose co-transporter 2 inhibitor empagliflozin (EMPA) by reducing the risk of incident or worsening nephropathy by 39% vs. placebo (PBO) in patients with T2DM and established cardiovascular (CV) disease. We investigated the effects of controlling the CV risk factors of blood pressure (BP), low-density lipoprotein cholesterol (LDL-C) and HbA1c on treatment differences in renal outcomes. Patients were randomized 1:1:1 to EMPA 10 mg, EMPA 25 mg, or PBO. Risk of incident or worsening nephropathy was assessed in the pooled EMPA group vs. PBO adjusting for control of BP, LDL-C and HbA1c at baseline and during the study as time-dependent covariates. Control of the various parameters was defined as systolic BP <140 mmHg and diastolic BP <90 mmHg, LDL-C <100 mg/dL, and HbA1c <7.5%. Adjusting for control of BP, LDL-C or HbA1c individually, hazard ratios (HR)s for time to incident or worsening nephropathy with EMPA vs. PBO ranged from 0.61 to 0.67 (Figure). Adjusting for control of all 3 parameters, the HR (95% CI) was 0.65 (0.57, 0.75) (Figure). EMPA reduced the risk of incident or worsening nephropathy to the same extent when analyses were adjusted for control of BP, LDL-C and HbA1c over time. These data suggest that risk reductions in renal outcomes were preserved irrespective of control of conventional CV risk factors.

Disclosure

C. Wanner: Board Member; Self; Boehringer Ingelheim GmbH, Sanofi Genzyme. M.E. Cooper: Advisory Panel; Self; AbbVie Inc.. Research Support; Self; Boehringer Ingelheim GmbH. Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc.. Speaker's Bureau; Self; Eli Lilly and Company. Advisory Panel; Self; Merck Sharp & Dohme Corp., AstraZeneca. Research Support; Self; Novo Nordisk A/S. Advisory Panel; Self; Novartis AG. S. Inzucchi: Other Relationship; Self; Boehringer Ingelheim Pharmaceuticals, Inc., AstraZeneca, Novo Nordisk Inc., Sanofi, Lexicon Pharmaceuticals, Inc.. Consultant; Self; VTV Therapeutics. Other Relationship; Self; Intarcia Therapeutics, Inc.. Consultant; Self; Janssen Pharmaceuticals, Inc., Alere Inc. B. Zinman: Consultant; Self; Novo Nordisk A/S, Boehringer Ingelheim Pharmaceuticals, Inc., AstraZeneca, Eli Lilly and Company, Janssen Pharmaceuticals, Inc., Sanofi, Merck & Co., Inc., Abbott. U. Hehnke: Employee; Self; Boehringer Ingelheim GmbH. M. von Eynatten: Employee; Self; Boehringer Ingelheim GmbH. A. Koitka-Weber: Employee; Self; Boehringer Ingelheim GmbH.
Urinary NGAL and KIM-1 Are Significantly Elevated in Young Adults (YA) with Type 1 (T1D) and Type 2 (T2D) Diabetes
Authors: SHEANON NICOLE M. , MOTTL AMY K. , DAGOSTINO RALPH , SUERKEN CYNTHIA , AFKARIAN MARYAM , DABELEA DANA , IMPERATORE GIUSEPPINA , MARCOVINA SANTICA M. , PETTITT DAVID J. , SAYDAH SHARON , DOLAN LAWRENCE M.
Published: June 2018

Urinary (U) neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) are markers of renal tubular injury. Whether NGAL and KIM-1 are elevated in early diabetic kidney disease (DKD) is not known. We performed a pilot study to compare the U concentrations (C) of NGAL and KIM-1 in YA with T1D or T2D from the SEARCH registry and healthy controls (n 45 in each group). The YA with diabetes were selected based on age at dx > 10 years and duration >8 years and were randomly selected across the quartiles of eGFR with similar age, gender and ethnicity. T1D (age 21 ± 2 years; duration 9.6 ± 1.0 years; HbA1c 9.3 ± 2.1% eGFR 126 ± 19 mL/min/1.73m2, UACR 11 ± 16 µg/mg), T2D (age 24 ± 3 years, duration 9.8 ± 1.2 years, HbA1c 8.5 ± 3.0%, eGFR 128 ± 21 mL/min/1.73m2, UACR 38 ± 71 µg/mg), controls (age 23 ± 3 years). In each group 50% were female, 41% white, 43% AA, and 16% Hisp. NGAL and KIM-1 were significantly elevated in T1D and T2D compared with controls (p < 0.001, p< 0.007). Although there was a trend for a greater UC of NGAL in T2D than T1D and a greater UC of KIM-1 in T1D than T2D, these differences were not statistically significant. Elevated UC of NGAL and KIM-1 are present in YA with T1D and T2D suggesting the presence of tubular damage early in the course of diabetes. Further investigation will determine if these markers can be used to assess the natural history of tubular injury or monitor therapeutic intervention in DKD.

Disclosure

N.M. Sheanon: None. A.K. Mottl: None. R. Dagostino: Consultant; Self; Teva Pharmaceutical Industries Ltd., Acelity, RedHill Biopharma, Edwards Lifesciences. C. Suerken: None. M. Afkarian: None. D. Dabelea: None. G. Imperatore: None. S.M. Marcovina: None. D.J. Pettitt: None. S. Saydah: None. L.M. Dolan: None.
Effect of Thiazolidinediones on Renal Outcomes in Diabetic Patients with Microalbuminuria or Macroalbuminuria-A Systematic Review and Meta-analysis
Authors: QIN YAO , WANG XING , ZHANG MEI
Published: June 2018

Introduction: Diabetic nephropathy (DN), has recently become the leading cause of end-stage renal disease (ESRD) in most countries, recent studies suggest that thiazolidinediones (TZDs) have renal benefits. The aim of this meta-analysis was to assess the effect of TZDs on renal outcomes in type 2 diabetic patients with microalbuminuria or macroalbuminuria.

Methods: Medline, Embase, Web of Science and Cochrane Central Register of Controlled Trials were searched from 1991 to September 2017. Randomized controlled trials (RCTs) were selected to evaluate the impact of TZDs on urinary albumin, protein excretion, eGFR or Scr in diabetic patients. Weighted (WMDs) and standardized mean differences (SMDs) were used for changes in urine albumin or protein excretion, eGFR and Scr between the TZDs and control groups.

Results: 26 RCTs (9 with rosiglitazone and 17 with pioglitazone) involving 8129 patients with type 2 diabetes were included in this review. Meta-analysis of RCTs showed that in patients with baseline normo- and/or microalbuminuria, the WMD of proportional changes between the TZDs and control groups in urinary albumin excretion was -64.77% (95% CI, -75.60 to -53.94) and the WMD of changes in albumin-creatinine ratio was -24.94% (95% CI, -38.83 to -11.05). Overall, in participants with normo- and/or microalbuminuria, TZDs treatment was associated with a significant decrease in urinary albumin excretion (SMD, -0.91 units of standard deviation [SD]; 95% CI, -1.19 to -0.64). Similarly, TZDs were associated with a significant decrease in urinary protein excretion in patients with macroalbuminuria (SMD, -1.units of SD; 95% CI, -1.86 to -0.27). However, TZDs treatment couldn’t effectively improve eGFR or the level of Scr in diabetic patients whether with microalbuminuria or macroalbuminuria.

Conclusions: Treatment with TZDs can benefit patients with early diabetic nephropathy in absence of renal dysfunction.

Disclosure

Y. Qin: None. X. Wang: None. M. Zhang: None.
Glycemic Fluctuation and the Risk of Hypoglycemia Unawareness Associated with Hemodialysis in Type 2 Diabetic Renal Failure
Authors: HAYASHI AKINORI , MOMOZONO AKARI , KAWAI SAYUKI , MASAKI TSUGUTO , YOSHINO SONOMI , OGAWA AKIFUMI , TAKANO KOJI , SHICHIRI MASAYOSHI
Published: June 2018

Despite the high-risk of asymptomatic hypoglycemia associated with hemodialysis (HD) using glucose-free dialysate, presence of dialysate glucose is believed to prevent intradialytic hypoglycemia. However, exact glycemic fluctuation profiles and frequency of asymptomatic hypoglycemia during HD remain unappreciated. Using continuous glucose monitoring, we evaluated glycemic excursions in 63 type 2 diabetic HD patients (44 male, HbA1c 6.4±1.2%) treated with a dialysate containing either 100, 125 or 150 mg/dL glucose. Average sensor glucose level (SGL) at start of HD after 7.2±5.0 U bolus insulin and breakfast was 187.9±56.2 mg/dL. SGL showed gradual and sustained decrease during HD irrespective of the dialysate glucose levels (Figure). SGL nadir reached below the dialysate glucose levels in 23 of 37 patients treated with dialysate containing 100 mg/dL glucose, in 14 of 17 patients with 125 mg/dL and in all 9 patients with 150 mg/dL. Fourteen of all 63 patients (22%) presented with HD-related hypoglycemia as defined by SGL <63 mg/dL during HD or immediately after HD up until the next meal. All hypoglycemic episodes revealed asymptomatic.

In conclusion, type 2 diabetic HD patients present a high-risk of HD-related hypoglycemia unawareness despite the use of dialysate containing 100-150 mg/dL glucose. SGL may fall well below the dialysate glucose level toward the end of HD.

Disclosure

A. Hayashi: None. A. Momozono: None. S. Kawai: None. T. Masaki: None. S. Yoshino: None. A. Ogawa: None. K. Takano: None. M. Shichiri: None.
Artificial Intelligence Predicts Progress of Diabetic Kidney Disease-Novel Prediction Model Construction with Big Data Machine Learning
Authors: MAKINO MASAKI , ONO MASAKI , ITOKO TOSHINARI , KATSUKI TAKAYUKI , KOSEKI AKIRA , KUDO MICHIHARU , HAIDA KYOICHI , KURODA JUN , YANAGIYA RYOSUKE , SUZUKI ATSUSHI
Published: June 2018

Background: Diabetic kidney diseases (DKD) including diabetic nephropathy is the most frequent cause of hemodialysis (HD), and more precise prediction model could be useful to early intervention of DKD.

Methods: We constructed new prediction model for DKD by using artificial intelligence (AI) based on electronic medical records (EMRs). From EMRs of 64,059 diabetes patients who visited our hospital, we extracted a variety of features. This model uses the stage of nephropathy as labels, and predicts whether the stage 1 patients will move up their stage after 180 days.

Results: AI constructed new prediction model by big data machine learning. First, AI extracted raw features in past 6 months at reference period, and selected 22 factors. Then, time series data analysis using convolutional autoencoder was conducted to find time series patterns relating to 6-month DKD aggravation. AI then constructed the prediction model with 17raw features as well as time series and text as secondary features using logistic regression. Finally, AI predicted DKD aggravation with 0.74 AUC score at maximum. Furthermore, DKD aggravation group had significantly higher incidence of HD than non-aggravation group in 10 years.

Conclusion: The new prediction model by AI could detect progress of DKD, which could contribute to more effective and accurate intervention to reduce HD.

Disclosure

M. Makino: Research Support; Spouse/Partner; THE DAI-ICHI LIFE INSURANCE COMPANY, LIMITED. Self. M. Ono: None. T. Itoko: Employee; Self; IBM. T. Katsuki: Employee; Self; IBM. A. Koseki: Employee; Self; IBM. M. Kudo: Employee; Self; IBM. K. Haida: Employee; Self; Daiichilife Insurance Company. J. Kuroda: Employee; Self; The Dai-ichi Life Insurance Company, Limited. R. Yanagiya: None. A. Suzuki: Research Support; Self; THE DAI-ICHI LIFE INSURANCE COMPANY, LIMITED.. Speaker's Bureau; Self; Astellas Pharma Inc., Mitsubishi Tanabe Pharma Corporation. Research Support; Self; EA Pharma Co Ltd, Daiichi Sankyo Company, Limited, Chugai Pharmaceutical Co., Ltd., Kyowa Hakko Kirin Co., Ltd., MSD K.K., Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Pfizer Inc., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited.