bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0511ENHANCER AND SUPER-ENHANCER DYNAMICS IN REPAIR AFTER ISCHEMIC ACUTE KIDNEY INJURY
Authors: Wilflingseder Julia , Willi Michaela , Lee Hye Kyung , Olauson Hannes , Jankowsky Jakub , Ichimura Takaharu , Erben Reinhold , Valerius M Todd , Hennighausen Lothar , Bonventre Joseph Vincent 
Published: June 2020

Abstract

Background and Aims
The endogenous repair process of the mammalian kidney allows rapid recovery after acute kidney injury (AKI) through robust proliferation of tubular epithelial cells. There is currently limited understanding of which transcriptional regulators activate these repair programs and how transcriptional dysregulation leads to maladaptive repair. Here we investigate the existence of enhancer dynamics in the regenerating mouse kidney.

Method
RNA-seq and ChIP-seq (H3K27ac, H3K4m3, BRD4, POL2 and selected transcription factors) were performed on samples from repairing kidney cortex 2 days after ischemia/reperfusion injury (IRI) to identify activated genes, transcription factors, enhancer and super-enhancers associated with kidney repair. Further we investigated the role of super-enhancer activation in kidney repair through pharmacological BET inhibition using the small molecule JQ1 in vitro and in acute kidney injury models in vivo.

Results
Response to kidney injury leads to genome-wide alteration in enhancer repertoire in-vivo. We identified 16,781 enhancer sites (H3K27ac and BRD4 positive, H3K4me3 negative binding) active in SHAM and IRI samples; 6,512 lost and 9,774 gained after IRI. The lost and gained enhancer sites can be annotated to 62% and 63% of down- and up-regulated transcripts at day 2 after kidney injury, respectively. Super-enhancer analysis revealed 164 lost and 216 gained super-enhancer sites at IRI day 2. 385 super-enhancers maintain activity before and after injury. ChIP-seq profiles of selected transcription factors based on motif analysis show specific binding at corresponding enhancer sites. We observed lost enhancer binding of HNF4A and GR mainly at kidney related enhancer elements. In contrast, STAT3 showed increased binding at injury induces enhancer elements. No dynamic was observed for STAT5. Both transcription factor groups show corresponding mRNA changes after injury. Pharmacological inhibition of enhancer and super-enhancer activity by BRD4 inhibition (JQ1: 50mg/kg/day) before IRI leads to suppression of 40% of injury-induced transcripts associated with cell cycle regulation and significantly increased mortality between days 2 and 3 after AKI.

Conclusion
This is the first demonstration of enhancer and super-enhancer function in the repairing kidney. In addition, our data call attention to potential caveats for use of small molecule inhibitors of BET proteins that are currently being tested in clinical trials in cancer patients who are at risk for AKI. Our analyses of enhancer dynamics after kidney injury in vivo have the potential to identify new targets for therapeutic intervention.
P0512SKLB023 ATTENUATED LIPOPOLYSACCHARIDE-INDUCED ACUTE KIDNEY INJURY VIA INHIBITION OF NF-KAPPPAB AND MAPKS PATHWAYS
Authors: Ren Qian , Ma Liang , Fu Ping 
Published: June 2020

Abstract

Background and Aims
Excessive nitric oxide (NO) production by the activation of inducible nitric oxide synthase (iNOS) during sepsis was considered to contribute to acute kidney injury (AKI) and selective inhibition of iNOS activity may be a promising strategy for the treatment of sepsis-induced AKI. The novel small molecule compound (Z)-N-(3-Chlorophenyl)-2-(4-((2,4-dioxothiazolidin-5-ylidene) methyl) phenoxy) acet-amide (SKLB023) as a selective iNOS inhibitor was designed, synthesized by our lab and exhibited therapeutic potential in arthritis and non-alcoholic steatohepatitis models with potent anti-inflammatory effects. This study aimed to evaluate whether SKLB023 as a drug candidate could offer renal protective effect against sepsis-induced AKI.

Method
21 C57BL6 mice (20-22g, 6-8 weeks old) were randomly divided into three groups: control, model, SKLB023 pretreatment. AKI was induced by a single injection of 10mg/kg LPS, and SKLB023 was orally administrated at the dose of 50 mg/kg for three consecutive days and one hour before LPS injection. All mice were sacrificed 16h after LPS injection and blood and kidney tissues were collected for further evaluation. LPS-induced renal dysfunction was assessed by measurement of blood creatinine and urea nitrogen (BUN) as well as histopathological changes. Moreover, the kidney mRNA and protein expression of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule 1(KIM-1), newly identified kidney injury markers, were tested by polymerase chain reaction (PCR) and western blot. To explore underlying mechanisms, the renal expression of inflammatory cytokine genes IL-1β, IL-6, mcp-1 and proteins iNOS, COX-2, IL-1β, TNF-α, IL-6, HMGB1 in were measured. The total expression of nuclear factor-kappa B (NF-κB), IκBα and MAPKs proteins (ERK1/2, p38 MAPK) and their corresponding phosphoproteins (p-NF-κB, p-IκBα, p-ERK1/2, p-P38 MAPK) in mice kidneys were subsequently analyzed by western blot.

Results
16hours following LPS challenge, mice developed AKI as evidenced by elevated creatinine and BUN levels and corresponding pathological changes. Additionally, kidney expression of NGAL and KIM-1 were remarkably increased after LPS injection. SKLB023 pretreatment, however, alleviated LPS-induced renal dysfunction. Moreover, the excessive production of inflammatory cytokines during sepsis was significantly suppressed by SKLB023. Further observation from PCR and immunoblot results indicated that SKLB023 attenuated the activity of NF-kB and the phosphorylation of ERK1/2 and p38 MAPK in septic AKI mice.

Conclusion
These data demonstrated that SKLB023 exhibited renoprotective effect in LPS-induced AKI, which was associated with its anti-inflammatory activities by inhibiting the NF-kappaB and MAPK signaling pathways
P0869EXOGENOUS OSTEOGENIC FACTOR 1 ADMINISTRATION ATTENUATED VASCULAR CALCIFICATION AND IMPROVED BONE DISORDERS IN CHRONIC UREMIC RATS
Authors: Lee Chien-Te , Ng Hwee-Yeong , Chen Wen-Lin , Lee Yuai-Ting 
Published: June 2020

Abstract

Background and Aims
Hyperphosphatemia and secondary hyperparathyroidism are frequent complications in chronic kidney disease (CKD) which both contribute to increased morbidity and mortality in CKD. Osteogenic factor-1 (OP-1) is an important member of BMPs subfamily and its effects on CKD-associated mineral and bone disorders (MBD) is controversial. The study examined whether exogenous OP-1 administration can modulate disturbed CKD-MBD in adenine-induced chronic uremic rats

Method
Chronic renal failure was induced in adult male SD rats by feeding adenine-containing diet. After adenine diet feeding 3 weeks, animals were injected with OP-1 (5μg/kg/day) intraperitoneally for 2 weeks. The serum and urine phosphorus levels and associated mineral parameters, including fibroblast growth factor 23(FGF-23), DKK-1 and sclerostin were measured. Vascular calcification was assessed by immunohistochemistry staining on aortic tissue. Bony structure was evaluated by microCT (Bruker-microCT, Kontich, Belgium).

Results
A significant decrease of body weight and deteriorated renal function was observed in adenine and OP-1 treatment groups during study period and serum creatinine levels were similar (5.23 ±1.1 mg/dL vs. 5.4 ±1.2 mg/dL, p&gt;0.05). Animals in OP-1 group had lower serum phosphorous (18.7±5.1 vs. 29.0±9.6 mg/dL, p&lt;0.05) and intact parathyroid hormone levels (2906.1±1206.9 vs. 4669.7±2505.9 pg/dL, p&lt;0.05) compared to adenine group. Decreased urine phosphorous excretion was noted in both groups without significant difference. Levels of serum FGF-23, sclerostin and DKK-1 were significantly lower in OP-1 treatment group (all p&lt; 0.05). OP-1 administration diminished the staining of RUNX2 (59.1±3% of adenine-treated group), alkaline-phosphatase (49.4±5.7%), β-caterin (39.3±1.8%), BMP2 (43.2%±6.7%), and BMP7 (51.9±10%, all p &lt;0.05). MicroCT revealed that bone mineral density was increased by OP-1 treatment (0.46±0.1 vs.0.39±0.06 g/cm3). Total volume was increased but bone volume was not changed. OP-1 administration did not affect trabecular thickness and trabecular number.

Conclusion
Our data indicated administration of exogenous OP-1 improved hyperparathyroidism and attenuated vascular calcification. OP-1 treatment was also associated with beneficial effects on bony structure in animals with renal failure.
P1371HEART FAILURE HOSPITALISATIONS IN THE PIVOTAL TRIAL OF IV IRON IN HAEMODIALYSIS PATIENTS: A PRE-SPECIFIED SECONDARY ANALYSIS
Authors: Jhund Pardeep , Petrie Mark , Robertson Michele , Mark Patrick , Macdonald Michael , Connolly Eugene , White Claire , Reid Chante , Anker Stefan D , Bhandari Sunil , Farrington Kenneth , Kalra Philip A , Tomson Charles R V , Wheeler David , Ford Ian , McMurray John , Macdougall Iain 
Published: June 2020

Abstract

Background and Aims
Heart failure (HF) is a common and potentially deadly complication in patients receiving haemodialysis which is difficult to diagnose and treat. The impact of a proactive high-dose strategy compared to a reactive low-dose strategy of IV iron administration was investigated in the PIVOTAL randomised trial in incident haemodialysis patients, with fatal HF and HF hospitalisation being components of the composite primary cardiovascular endpoint as well as a pre-specified key secondary event.
The aim of this analysis was to examine the effect of a proactive high-dose strategy compared to a reactive lower dose strategy on HF events in patients recruited to PIVOTAL.

Method
As with the primary composite cardiovascular endpoint, HF hospitalisations in PIVOTAL were recorded prospectively via an electronic database, and events were then adjudicated by a blinded Endpoint Adjudication Committee. The time-to-event analyses of the primary, secondary and post hoc outcomes were performed in the intention-to-treat population using Cox proportional hazards regression, with treatment group as the only explanatory variable. The Kaplan–Meier method was used to estimate event rates. Both fatal and non-fatal HF events were analysed as time to first event, and a recurrent event analysis was also performed for non-fatal events.

Results
Overall, 2141 participants were followed for a median of 2.1 years. A first fatal or non-fatal HF event occurred in 51 of 1093 patients (4.7%) in the high-dose iron group and in 70 of 1048 patients (6.7%) in the low-dose iron group (hazard ratio 0.66, 95% confidence interval 0.46 to 0.94; P&lt;0.001) (Figure). There was a total of 63 HF events (including first and recurrent events) in the high-dose iron group and 98 in the low-dose iron group, giving a rate ratio of 0.59 (0.40-0.87); p=0.0084. A history of HF and diabetes were independent predictors of a heart failure event.

Conclusion
Compared with a low-dose regimen, high-dose intravenous iron decreased the occurrence of first and recurrent heart failure events in incident patients undergoing haemodialysis, with large relative and absolute risk reductions.
P0338CRUMBS HOMOLOG2-EZRIN SIGNALING INDUCED PODOCYTE INJURY, LEADING TO MINIMAL-CHANGE DISEASE(MCD) AND FOCAL SEGMENTAL GLOMERULOSCLEROSIS(FSGS)
Authors: Hada Ichiro 
Published: June 2020

Abstract

Background and Aims
The etiology and cellular pathogenesis of podocyte injury leading to minimal-change disease (MCD) and focal segmental glomerulosclerosis (FSGS) remain largely obscure. Genetic mutation of crumbs homolog 2 (CRB2) is a cause of congenital nephrotic syndrome. Type-1 transmembrane proteins including CRB2 transduce outside-in signals that are involved in various cellular events including changes in the cytoskeletal network. The aim of the present study is to determine whether alteration of CRB2-mediated signaling in podocytes causes MCD and FSGS.

Method
Mice were immunized with a partial recombinant protein including the extracellular part of mouse CRB2. Urinalysis was obtained, and the kidney was subjected to histopathology. Kidney samples were also subjected to immunofluorescence microscopy and glomerular isolation to determine whether activation of the ezrin/radixin/moesin (ERM) family of cross-linkers between plasma membrane proteins and the actin cytoskeleton is involved in the pathogenesis of this nephrotic model. A CRB2-expressing mouse podocyte cell line was generated and incubated with anti-CRB2 antibody, and cell lysates were subjected to immunoblot analysis of ERM phosphorylation. The presence of anti-CRB2 antibody in the serum was determined by Western blot analysis.

Results
Apparent anti-CRB2 antibody was detected in the serum from 4 weeks onward. Immunized mice developed proteinuria at 4 weeks, which continued at least until 29 weeks. Mice developing extremely heavy proteinuria also developed hematuria from 18 weeks onward. Light microscopy revealed MCD in mice with proteinuria alone and FSGS in mice with heavy proteinuria and hematuria. Immunofluorescence microscopy revealed positive granular IgG staining in podocyte foot processes, but not complement C3. Electron microscopy and immuno-electron microscopy revealed alteration of actin organization associated with prominent foot process effacement. Strong phosphorylation of ezrin was observed in the glomerulus from the proteinuric stage and in the cellular lysates from the CRB2-expressing podocyte cell line incubated with anti-CRB2 antibody.

Conclusion
The current data revealed that binding of anti-CRB2 antibody to the extracellular domain of CRB2 on the podocyte foot process activated the ezrin-cytoskeleton network, leading to podocyte injury. Our data also indicated that signaling by this one molecular can induce two different phenotypes of glomerular injury: MCD and FSGS. In our model, the signaling was activated by anti-CRB2 antibody, but in patients with nephrotic syndrome the CRB2 ligands remain unknown. Therefore, it will be important to identify the soluble factors interacting with CRB2, which may be novel factors contributing to the pathogenesis of MCD and FSGS.
P0768NOVEL URINARY PROTEIN BIOMARKERS FOR UROTHELIAL CANCER IN PATIENTS WITH CHRONIC KIDNEY DISEASE
Authors: Huang Chiu-Ching , Chou Che-Yi , Shu Kuo-Hsiung , Chen Hung-Chun , Wang Ming-Chang , Chang Chia-Chu , Hsu Bang-Gee , Wu Mai-Szu , Yang Yuan-Lung , Chen Chao-Jung 
Published: June 2020

Abstract

Background and Aims
Introduction: Chronic kidney disease (CKD) patients are known to have greater risk of urothelial cancer (UC) than general population. Urine is the most non-invasive examination and is directly linked to UC. Urinalysis and urine cytology are the major screening tools; however, these tests have low sensitivity or specificity in early disease. In this study, we used both normal and Non-UC CKD patients as controls to discover novel and more specific protein biomarkers in urine for early UC diagnosis.

Aims
To discover novel and more specific urinary protein biomarkers for screening of urothelial cancer (UC) in CKD patients.

Method
This is a multicenter prospective cohort study, carried out at 10 nationwide hospitals covering different regions of Taiwan from 2013-2018. Subjects were Taiwanese healthy adults (healthy controls, n=210), patients with CKD but without UC (disease controls, n=273) and patients with ongoing UC (n=211). For non-UC patients, urine and blood specimens were collected at the time of the structured interview. For UC patients, urine and blood samples were collected before operation. iTRAQ-LC-MS/MS approach was applied to compare protein expression among normal, CKD and UC cohorts.

Results
A total of 3838 proteins in urine were identified and 2497 proteins of them can be quantified. By filtering with fold changes, published papers and Human ATLAS, 27 candidates were selected as potential biomarkers and were validated with ELISA assay in a large sample size of urine specimens. Among the 27 candidates, we found BRDT, CYBP, GARS and HDGF were highly-expressed in the UC cohort when compared to normal and CKD cohorts (Figure 1). To increase the diagnostic value of UC, we combined ROC of our newly discovered 4 protein biomarkers. The AUC values of our newly discovered 4-biomarker panel were higher than the AUC values of the published biomarker panel, including the FDA-approved protein biomarkers (Figure 2).

Conclusion
We discover and validate a highly specific urinary protein biomarker panel (BRDT, GARS, HDGF and CYBP) to distinguish UC from CKD and healthy controls. It may be used for high throughput screening of subjects, such as CKD patients, who are at high risk to develop UC.
P0966PLACENTAL GROWTH FACTOR DEFICIENCY AGGRAVATES DIABETIC NEPHROPATHY
Authors: Lim Ji Hee , Kim Yaeni , Kim Eun Nim , JIN YONGJIE , Kim Hyung Duk , Ban Tae Hyun , Choi Bum Soon , Park Cheol Whee 
Published: June 2020

Abstract

Background and Aims
Placental growth factor (PlGF) is a member of the vascular endothelial growth factor (VEGF) family. PlGF exerts favorable angiogenetic and lymphangiogenic activity by binding to VEGF-R1 and -R3, respectively. Due to its functional synergy with VEGF-A, it is required for a correct neovascularization during pathological conditions while inactivation of PlGF contributes to decrease of pathological angiogenesis. Because reduced angiogenesis and lymphagiogenesis that contribute to defective lipid drainage are implicated in the progression diabetic kidney disease, we investigated the role of PlGF in the development of diabetic nephropathy by using PlGF-knockout mice.

Method
Diabetes was induced by a low-dose streptozotocin injection in 9-week-old male C57BL/6J PlGF-KO and wild-type mice and biochemical and morphological parameters were examined at 12 weeks later.

Results
In diabetic PlGF-KO mice, fasting blood glucose and HbA1c levels increased significantly and the development of glomerular sclerosis and mesangial area expansion were accompanied by albuminuria. Diabetic PlGF-KO mice exhibited increased expression of type IV collagen, transforming growth factor-β1 and glomerular IHC staining for nephrin, PECAM-1 and WT-1-positive cells and VEGF-R1,-R2,-R3 expression decreased, suggesting decreased endothelial cell and podocyte structure. Intrarenal expression of pLKB1, and pAMPK decreased and that of PPARα, PGC1α, ERRα, p-eNOS, and urinary Nox concentration decreased while iNOS increased, indicating disturbed lipid metabolism and endothelial dysfunction in the same group. Increased intrarenal FFA, TG, and cholesterol concentration represents presence of lipid accumulation. F4/80- and TUNEL-positive cells increased, suggesting increased inflammatory cell infiltration and apoptosis. CD68 and arginase-II increased indicating that macrophage subtype M1 polarization is involved in the inflammatory process. Expression of Bcl2/bax decreased and that of SOD1 and 2 decreased, indicating increased apoptosis and oxidative stress, respectively. Increased expression of intrarenal 8-OHdG and urinary isoprostane level indicates increased oxidative stress.

Conclusion
Impaired angiogenesis and lymphangiogenesis are implicated in PlGF deficiency and this promotes lipotoxicity-induced inflammation, oxidative stress and deteriorates renal functional and phenotypic parameters in the diabetic kidney.
P1000URINE MYO-INOSITOL AS A NOVEL PROGNOSTIC BIOMARKER FOR DIABETIC KIDNEY DISEASE
Authors: Lim Chun Soo , Kwon Soie , Lee Jung Pyo 
Published: June 2020

Abstract

Background and Aims
Diabetic kidney disease (DKD) is a leading cause of chronic kidney disease (CKD) and end-stage kidney disease (ESRD). Metabolomics has been increasingly applied to identify the cause of CKD, as it can present epigenetics and suggest corresponding treatment options. Only a few metabolomics studies were conducted in DKD patients, and the results are inconclusive. We investigated the association between urine metabolites and renal disease progression and mortality in DKD, using 800 MHz NMR spectroscopy based targeted metabolomics profiling.

Method
Prospectively stored urine samples from consecutive patients with DKD stage 1 to 5 (n=208) and their healthy controls (n=26) were analyzed. Cross-sectional associations were evaluated between eGFR or urine protein creatinine ratio (UPCR) and 26 urinary metabolites. Multivariable adjusted Cox models were conducted for the risk prediction of ESRD and mortality. The receiver-operating characteristic (ROC) analyses were used to assess the additive effect of each metabolite to predict the progression to ESRD.

Results
ESRD occurred in 103 (44.0%) patients and 65 (27.8%) deaths occurred during median 4.5 years (IQR, 2.06–6.58) follow-up period. The median fold change in 9 metabolites (glucose, mannose, myo-inositol, glycerol, lactate, fumarate, creatine, taurine and choline) in patient group revealed a trend corresponding to DKD stages. Linear regression showed that myo-inositol had a strongest association with eGFR. The relationship between the competitive metabolites and outcomes (ESRD and mortality) was investigated by multivariate Cox models after adjusting for the baseline covariates. Of these, 4 metabolites (myo-inositol, glycerol, fumarate, oxoisocaproate) had predictive values for ESRD, and among them, only myo-inositol retained predictive significance in mortality (adjusted HR 1.004, 95% confidence interval 1.002-1.006, P &lt; 0.001). At ROC analysis, urinary myo-inositol had additive effect to serum creatinine concentration and UPCR in the prediction for ESRD progression (NRI = 2.9%, P = 0.03; IDI = 35.1%, P = 0.02).

Conclusion
These results suggest that myo-inositol can be a predictive biomarker for the risk of ESRD progression and mortality in DKD. Further studies are needed to elucidate the pathophysiological roles of myo-inositol in DKD.
P1431MULTI-CENTER REGISTRY ANALYSIS COMPARING SURVIVAL ON HOME HEMODIALYSIS AND KIDNEY TRANSPLANT RECIPIENTS IN AUSTRALIA AND NEW ZEALAND
Authors: Ethier Isabelle , Cho Yeoungjee , Hawley Carmel , Pascoe Elaine M , Roberts Matthew A , Semple David , Nadeau-Fredette Annie-Claire , Wong Germaine , Lim Wai , Sypek Matthew P , Viecelli Andrea , Campbell Scott , Van Eps Carolyn , Isbel Nicole M , Johnson David W 
Published: June 2020

Abstract

Background and Aims
In the era of organ shortage, home hemodialysis (HHD) has been identified as the possible preferential bridge to kidney transplantation. Data are conflicting regarding the comparability of HHD and transplantation outcomes. This study aimed to compare patient and treatment survival between HHD patients and kidney transplant recipients.

Method
The Australia and New Zealand Dialysis and Transplant (ANZDATA) Registry was used to include incident HHD patients on day 90 after initiation of kidney replacement therapy and first kidney-only transplant recipients in Australia and New Zealand from 1997 to 2017. Survival times were analyzed using the Kaplan-Meier product limit method comparing HHD patients to subtypes of kidney transplant recipients using the log-rank test. Adjusted analyses were performed with multivariable Cox proportional hazards regression models for time to all-cause mortality. Time-to-treatment failure or death was assessed as a composite secondary outcome.

Results
The study compared 1411 HHD patients to 4960 living donor (LD) recipients, 6019 standard criteria donor (SCD) recipients and 2427 expanded criteria donor (ECD) recipients. While LD and SCD recipients had reduced risks of mortality compared to HHD patients (LD adjusted hazard ratio [HR] 0.57, 95%CI 0.46-0.71; SCD HR 0.65 95%CI 0.52-0.79), the risk of mortality was comparable between ECD recipients and HHD patients (HR 0.90, 95%CI 0.73-1.12). LD, SCD and ECD kidney recipients each experienced superior time-to-treatment failure or death compared to HHD patients.

Conclusion
This large registry study showed that kidney transplant offers a survival benefit compared to HHD but that this advantage is not significant for ECD recipients.