Differential proteomic analysis of cyclosporine A-induced toxicity in renal proximal tubule cells

Puigmulé, Marta; López-Hellı́n, Joan; Suñé, Guillermo; Tornavaca, Olga; Camaño, Sonia; Tejedor, Alberto; Meseguer, Anna

doi:10.1093/ndt/gfp149

Cited by 32 publications

(27 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2). This corresponds to other studies describing effects after 24 h of CsA treatment in cell culture [8,14,23]. Accelerated apoptosis contributes to CsA associated interstitial fibrosis, an important feature of chronic CsA nephrotoxicity [24].…”

Section: Discussionsupporting

confidence: 86%

Cyclosporin-A Induced Toxicity in Rat Renal Collecting Duct Cells: Interference with Enhanced Hypertonicity Induced Apoptosis

Schenk

Rinschen²,

Klokkers³

et al. 2010

Cell Physiol Biochem

View full text Add to dashboard Cite

Background/ Aims: Rat renal inner medullary collecting duct (IMCD) cells are physiologically exposed to a wide range of ambient tonicity. To maintain their function upon changes in osmolality, IMCD cells induce expression of osmoprotective and antiapoptotic genes, mainly mediated by the transcription factor Tonicity Enhancer Binding Protein (TonEBP). Some drugs like Cyclosporin-A (CsA) are discussed to interfere with the activity of TonEBP and thereby mediate their nephrotoxic effects. The aim of our study was to further understand CsA toxicity during elevation of ambient osmolality. Methods: First we examined cytotoxicity of CsA in IMCD exposed to elevated tonicity. Employing microarray analysis of gene expression, real-time PCR and immunoassays, we scrutinized pathways contributing to this effect. Results: We show that in IMCD cells CsA but not FK506 increases apoptosis upon an increase in tonicity. This effect is independent of cellular TonEBP localization or activity and reactive oxygen species. Microarray studies revealed marked quantitative differences in gene expression. Functional analysis showed overrepresentation of genes associated with cell death in presence of CsA. This correlated with increased mRNA expression of genes associated with the death receptor pathway and detection of TNFΑ in culture medium of cells treated with CsA. Conclusion: Our results show that CsA cytotoxicity is induced under elevated ambient osmolality and that death receptor signaling probably contributes to CsA cytotoxicity.

show abstract

Section: Discussionsupporting

confidence: 86%

Cyclosporin-A Induced Toxicity in Rat Renal Collecting Duct Cells: Interference with Enhanced Hypertonicity Induced Apoptosis

Schenk

Rinschen²,

Klokkers³

et al. 2010

Cell Physiol Biochem

View full text Add to dashboard Cite

show abstract

“…Table 2 lists the 53 transcripts that have been previously reported to play a role in ischemia-reperfusion (IR), nephrotoxicity, or proteinuria [4,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45], suggesting a striking overlap in the kidney's transcriptional responses to a wide variety of insults. In the tubules, there was upregulation of many cell cycle, cell stress, apoptosis and inflammatory genes with concurrent downregulation of membrane transporters.…”

Section: Resultsmentioning

confidence: 99%

Gene Expression Analysis and Urinary Biomarker Assays Reveal Activation of Tubulointerstitial Injury Pathways in a Rodent Model of Chronic Proteinuria (Doxorubicin Nephropathy)

Cianciolo

Yoon²,

Krull³

et al. 2013

Nephron Exp Nephrol

View full text Add to dashboard Cite

Background: Tubular atrophy and interstitial fibrosis are well-recognized sequelae of chronic proteinuria; however, little is known regarding the molecular pathways activated within tubulointerstitium in chronic proteinuric nephropathies. Methods: To investigate the molecular mechanisms of proteinuria-associated tubulointerstitial (TI) disease, doxorubicin nephropathy was induced in rats. Progression of disease was monitored with weekly urinary biomarker assays. Because histopathology revealed multifocal TI injury, immunodirected laser capture microdissection was used to identify and isolate injured proximal tubules, as indicated by kidney injury molecule-1 immunolabeling. Adjacent interstitial cells were harvested separately. Gene expression microarray, manual annotation of gene lists, and Gene Set Enrichment Analysis were performed. A subset of the regulated transcripts was validated by quantitative PCR and immunohistochemistry. Results: Severe proteinuria preceded tubular injury biomarkers by 1 week. Histology revealed multifocal, mild TI damage at 3 weeks, which progressed in severity at 5 weeks. Affymetrix microarray analysis revealed tissue-specific regulation of gene expression. Manual annotation of gene lists, gene set enrichment analysis, and urinary biomarker assays revealed similarities to pathways activated in direct TI injuries. This suggests commonalities amongst the molecular mechanisms of TI injury secondary to proteinuria, ischemia-reperfusion, and nephrotoxicity.

show abstract

“…Contrary to cytoskeletal proteins, cell adhesion molecule phosphorylation is one of the most common molecular functions of downregulated proteins in response to TGF-β. As one of the key events of EMT is loss of epithelial adhesion properties [23], downregulation of cell adhesion molecule phosphorylation is consistent with phenotypic conversion of proximal epithelial cells in response to TGF-β.…”

Section: Discussionmentioning

confidence: 99%

“…Currently, the standard approaches to proteomics have been gel-based techniques (one/two dimensional gel electrophoresis) and gel-free techniques (liquid chromatography (LC)-based techniques) followed by mass spectrometry [7]. In renal research, gel-based technology has enabled us to identify new biomarkers in clinical nephrology [15,16,17,18,19,20], explore mechanism of the diseases [18,21,22,23,24,25] and identify new therapeutic targets [20,26]. However, during the past few years, gel-based techniques have been challenged and complemented by LC-based techniques because of its poor representation of low abundant proteins, highly acidic/basic proteins, or proteins with extreme size or hydrophobicity [27,28,29].…”

Section: Discussionmentioning

confidence: 99%

Phosphoproteomic Study of Human Tubular Epithelial Cell in Response to Transforming Growth Factor-Beta-1-Induced Epithelial-to-Mesenchymal Transition

Chen

Wang

et al. 2009

Am J Nephrol

View full text Add to dashboard Cite

Background: Transforming growth factor-β (TGF-β)-induced epithelial-to-mesenchymal transition (EMT) plays an important role in renal fibrosis and progression of chronic kidney disease (CKD). Phosphorylation of proteins is essential to TGF-β signaling. We applied isobaric tags for relative and absolute quantification (iTRAQ) technology to profile the phosphoproteins in tubular epithelial cells in response to TGF-β-induced EMT in order to further study molecular events. Methods: HK-2 cells were treated with TGF-β1 to induce EMT. The cells were divided into a control group (without TGF-β1 treatment) and a TGF-β1-treated group. Phosphoproteins from two groups were extracted and differentially labeled with iTRAQ reagents and processed by 2D-nano-HPLC-MS/MS. Validating of iTRAQ analysis was performed by western blot. Bioinformatic analysis was performed by on-line databases. Results: By iTRAQ-2D-nano-HPLC-MS/MS, 38 differentially expressed phosphoproteins were identified which included 19 up-regulated phosphoproteins and 19 down-regulated phosphoproteins. Western blot confirmed up-regulation of phosphorylated moesin and HSP90α. Bioinformatic analysis suggested that the majority of proteins were located in the nucleus and endoplasmic reticulum lumen. The phosphoproteins were categorized into 17 molecular function classifications. Nucleic acid binding protein, cytoskeletal protein and chaperone were the major categories of molecular function. A biological network was built to analyze interaction between up-regulated proteins. Conclusion: We demonstrate a TGF-β1-mediated post-transcriptional regulation of EMT in tubular epithelial cells. Phosphorylation of moesin and HSP90α might play a role in TGF-β-induced EMT.

show abstract

Differential proteomic analysis of cyclosporine A-induced toxicity in renal proximal tubule cells

Cited by 32 publications

References 49 publications

Cyclosporin-A Induced Toxicity in Rat Renal Collecting Duct Cells: Interference with Enhanced Hypertonicity Induced Apoptosis

Cyclosporin-A Induced Toxicity in Rat Renal Collecting Duct Cells: Interference with Enhanced Hypertonicity Induced Apoptosis

Gene Expression Analysis and Urinary Biomarker Assays Reveal Activation of Tubulointerstitial Injury Pathways in a Rodent Model of Chronic Proteinuria (Doxorubicin Nephropathy)

Phosphoproteomic Study of Human Tubular Epithelial Cell in Response to Transforming Growth Factor-Beta-1-Induced Epithelial-to-Mesenchymal Transition

Contact Info

Product

Resources

About