bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0041CHARACTERISTICS OF URINE-DERIVED RENAL EPITHELIAL CELLS (UREC) FROM CHILDREN WITH POLYCYSTIC KIDNEY DISEASE (PKD)
Authors: Ziegler Wolfgang H , Lüdiger Sarah , Mett Charlotte , Hasan Fatema , Soetje Birga , Haffner Dieter 
Published: June 2020

Abstract

Background and Aims
In hereditary PKD, epithelial cell defects are mostly caused by mutation of proteins of the cilia-centrosome complex. Use of primary cells from patients allows to study specific cell characteristics and altered activation of signal pathways in epithelial cell function-related culture models. In this ex vivo approach, comparison of cell properties is based on defined genetic origin and can be related to kidney function at the time of UREC collection, revealing consequences of genetic disposition and kidney-stress related changes on cell function.

Method
The study considers patients with genetically confirmed causes of PKD, autosomal recessive polycystic kidney disease (ARPKD), nephronophtisis (NPH), Bardet-Biedl syndrome (BBS), and age-matched controls with normal kidney function. UREC are grown from cellular urine sediment using selective growth conditions. Primary cells are cultured up-to 21 days (passages 2 & 3) and tested with respect to their proliferation, tubular origin and epithelial properties in 2D/3D culture conditions. To allow pathway analysis and improve control conditions, immortalized UREC lines are also included.

Results
UREC preparations of cohorts from ARPKD, NPH, and BBS patients, and controls were studied quantitatively to determine specific epithelial cell properties. Considering basic characteristics of each primary culture including cell morphology, expression of epithelial markers, and formation of cell-cell adhesions, the potential of UREC cells to polarize and generate liquid-filled epithelial spheroids was measured. The capacity of lumen formation in 3D culture strongly varies among individual cell preparations and appears to reflect the genetic origin of UREC cells, whereas no correlation to (remaining) kidney function (eGFR) is observed.

Conclusion
Determining specific, quantitative UREC cell properties related to genetics and kidney function, we expect to gain a better mechanistic understanding of cellular and renal epithelial defects in PKD patients. Ex vivo analysis based on UREC cells may provide options for testing of personalized pharmaceutical intervention.
P0042SPECTRUM OF RENAL INVOLVEMENT IN CHILDREN WITH HNF1B NEPHROPATHY
Authors: Papizh Svetlana , Prikhodina Larisa 
Published: June 2020

Abstract

Background and Aims
Hepatocyte nuclear factor 1 gene (HNF1B) encodes for HNF-1ß, a transcription factor that is expressed early in embryonic development and plays a key role for tissue-specific regulations of gene expression in various organs such as the kidneys, urogenital tract, liver, biliary ducts and pancreas. HNF1B disease (MIM 137920) is recognized to represent an autosomal dominant syndromal disorder comprising variable phenotype with renal and extrarenal manifestations. Renal phenotype in HNF1B nephropathy is highly heterogeneous. The aim of the study was to evaluate the clinical characteristics of renal involvement in children with genetically proven HNF1B nephropathy.

Method
Five children (2M/3F) from unrelated families with heterozygous HNF1β mutations and kidney disorders were examined. We evaluated serum levels of electrolytes, magnesium (Mg), phosphorus (P), uric acid, acid-base composition; urinary excretion of Ca, P, creatinine (Cr); fractional excretion (FE) of Mg, urate (Ur), Na, K; tubular maximum phosphate reabsorption per GFR (TmP/GFR), and urinary β-2 microglobulin. Molecular genetic analysis was performed using by next generation sequencing.

Results
Cortical or medullary bilateral renal cysts were detected in all five children with HNF1B nephropathy. Kidney hypoplasia, medullary nephrocalcinosis, and low-grade proteinuria (&lt;0.5 g/m2/d) revealed in 2\5 (40%) of cases. One child had vesicoureteral reflux. All patients with HNF1B nephropathy had normal serum level of uric acid and all electrolytes, including Mg. Increased FE of Mg and urinary excretion of β-2 microglobulin was found in all cases. Increased FE of Ur, Na and K revealed in 4\5 (80%) of subjects. Decreased TmP/GFR had 3\5 (60%) of cases. At the first examination of patients aged 1.0 (IQR: 1.0; 2.0) years all of them already had decreased eGFR to CKD stage 2-3.

Conclusion
The present study demonstrates that patients with HNF1B-associated disease had wide spectrum of renal involvement. The most prevalent features of HNF1B nephropathy in our children were bilateral renal cysts, defects of proximal tubular function with wasting of electrolytes and progression to CKD stage 2-3 at early age.
P0049IDENTIFYING HUB GENES ASSOCIATED WITH CLINICAL CHARACTERISTICS IN IGA NEPHROPATHY BYWGCNA
Authors: Yang Chengyu , Xu Yan 
Published: June 2020

Abstract

Background and Aims
Clinically, IgA nephropathy has a variety of symptoms including paroxysmal gross hematuria, nephritic syndrome and nephrotic syndrome. This study aimed at investigating hub geneand genes modular related to IgA nephropathy clinical characteristics by using weighted gene co-expression network analysis combining clinical, microarray and network database parameters.

Method
We collected 32 human samples from the European Renal cDNA Bank and used RMA method to preprocess the raw data and utilize the limma package to obtain differentially expressed gene in renal interstitium and glomeruli. And then, we used the WGCNA package to construct the gene co-expression of differential expression genes and identify the hub genes associated with clinical characteristics in renal interstitium and glomeruli, respectively. Gene ontology enrichment analysis and KEGG analysis for hub genes which associated with clinical characteristics were performed by DAVID, and PPI information was acquired from STRING with visualization by Cytoscape.

Results
For glomeruli, 1470 genes differentially expressed between IgA nephropathy patients and healthy control, containing 10 hub genes associated with age, 8 hub genes associated with sex, 48 hub genes associated with Bp enrichd in ERK1 and ERK2 cascade and Rap1 signaling pathway, 223 hub genes associated with BMI enrich in organic acid catabolic process and fatty acid degradation pathway, 136 hub genes associated GFR enriched in immune response and PI3K-Akt signaling pathway, 82 hub genes associated with proteinuria enriched in extracellular matrix organization and PI3K-Akt signaling pathway. In tubulointerstitium, there were 480 genes differentially expressed between IgA nephropathy patients and healthy control. Among 480 DEGs, 6 hub genes associated with age, 15 hub genes associated with sex, 35 hub genes associated with Bp enrichd in positive regulation of apoptotic process, 87 hub genes associated with GFR enriched in negative regulation of macromolecule metabolic process and RNA transport, 33 hub genes associated with proteinuria enriched in regulation of apoptotic process and FoxO signaling pathway. PPI enrichment analysis shown that all hub genes sets are biologically connected cluster.

Conclusion
We made a preliminary investigation on molecular mechanisms of relationship between IgA nephropathy and clinical characteristics and identified hub genes and pathways closely related with BMI, GFR and Proteinuria in IgA nephropathy by a series of bioinformatics analysis.
P0061LOSS OF UBIQUITIN SPECIFIC PROTEASE-40 CAUSES PODOCYTE DETACHMENT AND CONTRIBUTES TO THE DISEASE PROGRESSION OF GLOMERULOSCLEROSIS
Authors: Mikami Naoaki , Tanaka Eriko , Hada Ichiro , Yan Kunimasa 
Published: June 2020

Abstract

Background and Aims
We reported that the ubiquitin specific protease-40 (USP40) is expressed in podocytes beyond the species and plays a crucial role in glomerulogenesis in zebrafish (Takagi H, Am J Physiol Renal Physiol 2017). However, the role of USP40 in podocytes and its functional downstream after birth is largely unknown. Recent report indicated that other USP family: USP10 regulates integrin beta1 and fibrotic wound healing in myofibroblasts (Gillespie SR, J Cell Sci 2017). The present study aimed to explore the biological role of USP40 in the acquired podocyte injury and to determine the interacting partner by focusing on podocyte detachment.

Method
We generated USP40-knockout mice (USP40KO); however, they showed no apparent kidney abnormality. Therefore, to explore the biological role of USP40 in acquired podocyte injury, we crossed USP40KO with NEP25 mice, in which selective podocyte injury can be induced by injection with an immunotoxin, LMB2. Urinary protein was analyzed until day 9 after LMB2 injection, and then kidney tissues were subjected to light microscopy, immunohistochemistry of p57, and immunofluorescence microscopy of integrin beta1. The glomeruli were isolated and subjected to immunoblot analysis of integrin beta1. The effect of USP40 knockdown on integrin beta1 expression was studied in cultured mouse podocytes. Finally, the protein binding of integrin beta1 with USP40 was determined by immunoprecipitation using cultured mouse podocytes.

Results
NEP-USP40KO mice developed earlier and higher levels of proteinuria compared with NEP mice. Light microscopy and immunohistochemistry showed higher levels of kidney injury in NEP-USP40KO mice compared with NEP mice in terms of hyalinosis (p&lt;0.01), crescent formation (p&lt;0.01), and cell proliferation (p&lt;0.01) in the glomeruli, podocyte loss (p&lt;0.05), and tubulointerstitial injury (p&lt;0.01). Immunoblot analysis and immunofluorescence revealed decreased expression of integrin beta1 in podocytes of NEP-USP40KO mice compared with NEP mice. Gene knockdown of USP40 in cultured podocytes decreased the protein expression of integrin beta1. Finally, apparent protein binding of USP40 with integrin beta1 was observed.

Conclusion
Integrin beta1 is known to play a crucial role as the adhesion molecule that connects the podocytes with the extracellular matrix of the glomerular basement membrane. It is also known that the lack of integrin beta1 causes podocyte detachment, which is involved in the disease progression of glomerulosclerosis. Therefore, the present study suggests that USP40 may be involved in a self-defense mechanism in podocyte injury probably through interacting with the regulatory machinery of integrin beta1 in podocytes.
P0062GLUCOSYLCERAMIDE SYNTHASE INHIBITION WITH VENGLUSTAT IN CLASSIC FABRY DISEASE PATIENTS LEADS TO PROGRESSIVE REDUCTION OF ENDOTHELIAL CELL GLOBOTRIAOSYLCERAMIDE INCLUSION VOLUME
Authors: Mauer Michael , Goker-Alpan Ozlem , Germain Dominique P , Wilcox William , Hopkin Robert J , Lukina Elena , Geberhiwot Tarekegn , Deegan Patrick , Tylki-Szymanska Anna , Sensinger Charlotte , Zaher Atef , Hailman Eric , Modur Vijay , Najafian Behzad 
Published: June 2020

Abstract

Background and Aims
Fabry disease (FD) is a rare, X-linked, genetic disorder caused by mutations in the gene for the lysosomal enzyme alpha-galactosidase A (αGal-A). The progressive accumulation of glycosphingolipids, most notably globotriaosylceramide (GL-3), in lysosomes of a variety of organs and cell types leads, over decades, to renal, cardiovascular, and other clinical manifestations. In a phase 2 study, glucosylceramide synthase inhibition with venglustat led to a progressive reduction in plasma GL-3 to low-normal levels. Scoring by light microscopy showed a reduction in lysosomal GL-3 inclusions in skin capillary endothelial cells (EC) after 3 years of therapy, although not after 6 months. To further characterize the effect of venglustat using more precise methods, we applied unbiased stereological methods to quantitate GL-3 inclusions in skin ECs.

Methods
Skin biopsies were obtained from classic male Fabry disease patients (N = 11) at baseline and during daily treatment with venglustat in a phase 2 study (NCT02228460, NCT02489344). Images from at least 50 superficial skin capillaries were obtained using transmission electron microscopy at 7,500 X magnification. The fraction of the volume (Vv) of cytoplasm occupied by GL-3 inclusions [Vv(Inc/Endo)] in all ECs in these capillaries was estimated using point counting by a masked reader. Two-sided paired t tests were used to evaluate differences between baseline and post-treatment values at each time point.

Results
Venglustat therapy led to a significant reduction from baseline in Vv(Inc/Endo) of 21.1% after 6 months and 38.7% after 3 years of treatment (Figure).

Conclusions
Treatment with venglustat led to a reduction in GL-3 inclusion fractional volume in skin capillary ECs which was detectable after 6 months using precise quantitative methods. This was followed by further reduction over the next 2 1/2 years. We posit that, in the absence of αGal-A activity, inhibition of GL-3 production with venglustat allowed other enzymatic or non-enzymatic mechanisms to progressively reduce EC lysosomal GL-3 content, and that long-term venglustat therapy may therefore prevent or reverse progressive tissue injury in Fabry disease.
P0078INVESTIGATION THE VARIANTS AT THE BINDING SITE OF INFLAMMATORY TRANSCRIPTION FACTOR IN PATIENTS WITH ESRD
Authors: SU SUI-LUNG 
Published: June 2020

Abstract

Background and Aims
Inflammation is an important factor for enhancing the disease process from chronic kidney disease (CKD) to end-stage renal disease (ESRD). Nuclear factor-kappa B (NF-κB) is a transcription factor that regulates the expression of genes involved in inflammation.
We investigated the potential association with the gene polymorphism of transcription factor binding site of NF-κB in ESRD patients.

Method
We used the Taiwan Biobank database, University of California, Santa Cruz, reference genome, chromatin immunoprecipitation sequencing database to find the SNPs at potential binding sites of NF-κB. In addition, we performed a case–control study and genotyped 847 patients with ESRD and 846 healthy controls at Tri-Service-General-Hospital from 2015 to 2016. Further we used ChIP-assay and Luciferase reporter assay to identify the binding activity at different genotype.

Results
Results of biometrics screening in the databases revealed 15 SNPs with the potential binding site of NF-κB. Genotype distributions of rs9395890 were significantly different in ESRD cases and healthy controls (P = 0.032). In the Dominant model, rs9395890 with T allele had a higher risk of ESRD (P = 0.032; odds ratio [OR] = 1.32, 95% confidence interval [CI] = 0.99–1.76). The ChIP assay reveals that around 1.49 times enrichment of NF-κB of the variant type TT when compared to that of the wild type GG in the rs9395890 (P&lt;0.027; TT=3.20±0.16, GT=2.81±0.20, GG=1.71±0.18,) and the luciferase activity curve showed T allele was higher than G allele.

Conclusion
In conclusion, we demonstrate that rs9395890 may be associated with ESRD in the Taiwanese population.
P0079STRATEGIC ACTION FOR POLYCYSTIC KIDNEY DISEASE IN GALICIA. POPULATION APPROACH MODEL EXPORTABLE TO OTHER AUTONOMOUS COMMUNITIES
Authors: Besada Cerecedo Lara , Arhda Nisrine , Barcia de la Iglesia Ana Maria , Garcia Murias Maria , Carrera Cachaza Noa , Investigadores GalCYST Consorcio , Garcia Gonzalez Miguel 
Published: June 2020

Abstract

Background and Aims
The Strategic Action for Polycystic Kidney Disease (PKD) in Galicia was started in 2016, as a population strategy model (genetic cascade study) coordinated between the reference hospitals in this region, with the objective of diagnosing cost-effectively Galician families with the disease

Method
We develop panels for the simultaneous study of the currently known candidate genes, using ultrasequencing technologies (NGS). Each index case is analyzed by NGS and in the rest of the family members, the presence / absence of the eventual mutation identified is analyzed by Sanger Sequencing.Patient clinical data were collected to establish the genotype-phenotype correlation

Results
NefroCHUS has a historical record of 224 Galician families with PKD. Since January 1, 2016, 306 new PKD families have been identified from hospitals belonging to the three Galician health areas: Coruña - Ferrol (CHUAC (67 families), CHUF (17 families)), Vigo - Ourense (CHOP (44 families), CHUOU (43 families), CHUVI (32 families) and Povisa (18 families)) and Santiago-Lugo (HULA (35 families), HCB (22 families) and CHUS (28 families)), for a total record of 530 PKD families identified based on clinical criteria. Carrying out genetic studies allowed the calculation of gene mutagenicity rates for each health area: Coruña-Ferrol (58% PKD1, 36% PKD2 and 6% GANAB), Vigo-Ourense (80% PKD1, 15% PKD2 and 5 % other genes) and Santiago-Lugo (53% PKD1, 44% PKD2 and 3% GANAB). Taking into account the families of the historical record, mutagenicity rates in the Galician population are distributed as follows: 70% PKD1, 28% PKD2 and 2% other genes. So far 446 patients have been phenotypically characterized. The average age of diagnosis was 38 years, the main reason being family history (59%; n = 264). 73% of the patients presented arterial hypertension (AHT) with an average detection age of 42 years (range 6-82 years). 21% of the patients (n = 92) iniciated renal replacement therapy with a mean age of 49 years (38-80 years). The presence of liver cysts proved to be a very frequent extrarenal manifestation (67.9%; n = 303). Other manifestations to highlight are: low back pain (27%; n = 119), cystic kidney infection (7.4%; n = 33) or the presence of intracranial aneurysms (3.1%; n = 14)

Conclusion
It has been established the genetic map of the Autosomal Dominant Polycystic Kidney Disease in Galicia and at the same time a model of population approach in hereditary diseases that is exportable to other autonomous communities.
P0083HLA-DRB1*1301-DQB*0603-DRB3*0202 HAPLOTYPE IS INDEPENDENTLY ASSOCIATES WIHT POOR PROGNOSIS IN PLA2R-RELATED MEMBRANOUS NEPHROPAHTY
Authors: Le Weibo , Shi JingSong , Hou Qin 
Published: June 2020

Abstract

Background and Aims
The susceptibility of Idiopathic membranous nephropathy (MN) is associated with HLA alleles. We previously found almost all (98.7%) patients with PLA2R-related MN carry either or both of the HLA-DRB1*1501 and DRB3*0202 risk alleles. However, the associations of HLA alleles and renal outcome is not known.

Method
we investigated the associations between HLA alleles and renal outcome in 392 PLA2R-related MN patients.

Results
We found that HLA-DRB1*1301 and DQB1*0603, which are in high linkage disequilibrium (R2 = 1.0, D’=1.0), are associated with at least 30% eGFR decline during follow-up (HR 2.2, p&lt;0.001). HLA-DRB1*1301 and DQB1*0603 are not a susceptible HLA allele for PLA2R-related MN. Neither HLA-DRB1*1501 nor DRB3*0202 is associated with renal outcome. After adjusting age, gender and the two susceptible HLA alleles, DRB1*1301 haplotype remains significant associated with renal outcome. Compared those patients with negative DRB1*1301, patients with positive DRB1*1301 showed a similar eGFR level at baseline, but a significant lower eGFR at the last follow up. There were 12 PLA2R-related MN patients with positive DRB1*1301, eight of them (66.7%) showed more than 30% eGFR decline during follow-up. Among the 12 patients carrying DRB1*1301, all (100%) had DRB3*0202, four (33.3%) had DRB1*1501, and none (0%) had DRB1*0301. Linkage disequilibrium analysis showed the susceptible allele DRB3*0202 was highly linked with DRB1*0301 and DRB1*1301, while HLA-DRB1*0301 was not associated with renal outcome.

Conclusion
HLA-DRB1*1301-DQB*0603-DRB3*0202 haplotype is independently associated with Poor Prognosis in PLA2R-related Membranous.
P0087TRAF3 MODULATION: A NOVEL MECHANISM OF THE ANTI-INFLAMMATORY EFFECTS OF VITAMIN D RECEPTOR AGONIST PARICALCITOL IN RENAL DISEASE
Authors: Rayego-Mateos Sandra , Morgado Pascual Jose Luis , Valdivielso José M , Sanz Ana , Bosch panadero Enrique , Rodrigues Diez Raul , Egido Jesus , Ortiz Alberto , Gonzalez Parra Emilio , Ruiz-Ortega Marta 
Published: June 2020

Abstract

Background and Aims
Tumor necrosis factor receptor-associated factors (TRAFs) are critical signaling adaptors downstream of pro-inflammatory receptors, involved in canonical and non-canonical nuclear factor-κB (NF-κB) activation. Vitamin D receptor agonists (VDRAs) exert beneficial effects in renal disease and possess anti-inflammatory properties, but the underlying mechanism remains unknown. Our aim was to investigate TRAF3 involvement on renal disease and its potential modulation by VDRAs.

Method
Studies were done in isolated peripheral blood mononuclear cells (PBMCs) from end-stage renal disease (ESRD) on haemodialysis (treated or not with VDRAs) patients, healthy donors or cultured renal cells. Preclinical models of renal damage were: unilateral ureteral obstruction (UUO), acute renal damage induced by Folic acid (FA) administration and TWEAK-induced renal inflammation. The effect of VDRA (paricalcitol) was tested in vitro and in vivo.

Results
In PBMCs isolated from ESRD, TRAF3 protein levels were downregulated compared to healthy controls, associated to activation of non-canonical NF-κB2 (downregulation of RelB or p52/NF-κB2 protein levels) and altered pro-inflammatory cytokine profile. Interestingly, treatment of those patients with the VDRA paricalcitol restored all the described features. In vitro experiments in PBMCs and tubular epithelial cells exposed to inflammatory stimuli showed that paricalcitol prevented TRAF3 downregulation, NF-κB2 activation and proinflammatory genes upregulation. Moreover, immunoprecipitation studies showed that paricalcitol diminished TRAF3 ubiquitination in the K48-linked chains and CIAP1-TRAF3 interaction. TRAF3 overexpression by CRIPS/cas9 technology mimics VDRAs effects. In preclinical models of renal damage paricalcitol inhibited renal NF-κB2 activation, reducing p52 and RelB nuclear accumulation and transcriptional activity associated to lower renal inflammation. In TWEAK-induced renal injury in VDR knockout mice, paricalcitol prevented TRAF3 downregulation and NF-κB2-dependent gene upregulation, suggesting a VDR-independent anti-inflammatory effect of paricalcitol.

Conclusion
The present studies identify TRAF3 downregulation as a key feature promoting inflammation in CKD identifying non-canonical NF-κB activation as a key driver of inflammation in this context. The fact that the VDR is not required for paricalcitol actions suggests novel-signaling pathways involved in the anti-inflammatory effect of paricalcitol. This may allow the design of drugs that restore TRAF3 levels but are devoid of VDR-dependent effects on mineral bone metabolism, which may limit the use of VDRAs as anti-inflammatory agents.