Effect of Cardiac LIM Protein (CSRP3) on human proximal tubule epithelial cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE276233 | GEO | 2025/08/20
REPOSITORIES: GEO
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Project description:Angiotensin II (AngII), the effector of the renin angiotensin system causes kidney disease progression by signalling through AT-1 receptor, but there are no measures of AngII activity in the kidney. Accordingly, we sought to define an AngII-regulated proteome in primary human proximal tubular epithelial cells (PTEC) in order to identify potential markers of AngII activity in the kidney. PTECs were labeled with SILAC heavy arginine(+6) and lysine(+8) OR light arginine and lysine for 6 doubling times. Heavy-labeled PTECs were stimulated with AngII and light-labeled PTECs were stimulated with the control medium. AngII-treated and control-treated PTEC lysates were mixed in 1:1 total protein ratio and their proteomes were compared. We generated 5 biological replicates (1 supernatant, 1 replicate with reverse labeling). LTQ-Orbitrap mass spectrometer was used for LC-MSMS analysis. Of 4618 quantified proteins, 83 were differentially regulated by AngII in 4 biological replicates of PTEC lysates. We subsequently confirmed and verified 18 differentially regulated proteins by using SRM, RT-PCR and ELISA. We also went on to validate the main functional, enriched networks by using systems biology approach and an in vivo mouse model.
2013-07-18 | PXD000183 | Pride
Project description:Acute cardiorenal syndrome (CRS-1) is a morbid complication of acute cardiovascular disease. Mechanistic investigations have focused on intrarenal cellular signaling induced by ischemia/reperfusion. Additional signals, “cardiorenal connectors”, have been postulated, but investigation in CRS-1 has been limited by a paucity of animal models and technical limitations precluding discovery studies of glomerular filtrate. To address these limitations we developed a translational model of CRS-1, cardiac arrest and cardiopulmonary resuscitation (CA/CPR) and now report findings from a nanoscale mass spectrometry assay allowing proteomic exploration of Bowman’s space aspirate 2h after CA/CPR or sham procedure. Imaging, molecular weight and charge distribution, and minimal contribution of proteins from surrounding cell types confirmed the acquisition of filtrate. We detected filtration of low-molecular weight proteins specific to the heart following CA/CPR. Additional mass spectrometry performed on 24h urine collections from mice with deficient tubular endocytosis confirmed CA/CPR-specific cardiac protein filtration, and identified a novel, CA/CPR-specific, filtrate component: Cardiac LIM protein. Cardiac arrest-induced plasma release of Cardiac LIM protein occurred in mice and in critically-ill human cardiac arrest survivors and administration of recombinant cardiac LIM protein to mice altered renal function. Our findings demonstrate that glomerular filtrate is accessible to nanoscale proteomics and elucidate the population of proteins filtered 2h after acute cardiovascular crisis. The presence and identification of cardiac-specific proteins in renal filtrate suggest a potential novel signaling mechanism in CRS-1. We expect these findings to advance understanding of cardiorenal syndrome.
2019-02-27 | PXD012148 | Pride
Project description:Diabetic kidney disease (DKD) is the leading cause of end-stage kidney disease (ESKD), and few treatment options are available today to prevent the progressive loss of renal function. Tubular abnormalities may precede glomerular pathology early and indicate the functional progression of DKD, however, pathological mechanisms that initiate tubular abnormalities are poorly understood. Here, we found that glucagon receptor (Gcgr) is specifically and highly expressed in proximal tubular cells (PTEC) of kidney. Glucagon injection exacerbated lipid accumulation, glycogen content, inflammation, fibrosis, and renal injury, along with morphology changes on proximal tubules, podocytes, glomerular basement membrane (GBM), and mitochondria in the early phase of DKD mice. Whereas, the specific knockdown or knockout of Gcgr in PTEC of kidney almost completely halted the development of DKD. In contrary to the effect of short-term glucagon stimulation on fatty acid oxidation, long-term glucagon exposure led to glucagon reversal in PTEC, which is characterized by reduced energy production and promoted lipogenesis, and this effect was through the Gcgr-PKA-Creb-mTOR pathway. Accordingly, anti-GCGR antibody treatment greatly blocked the pathogenesis of DKD induced by both type 2 and type 1 diabetes. Thus, our results revealed a novel role of glucagon/GCGR signaling in PTEC lipogenesis and DKD, and Gcgr would be a promising therapeutic drug target for the treatment of DKD.
2025-07-18 | GSE253774 | GEO
Project description:Acute cardiorenal syndrome (CRS-1) is a morbid complication of acute cardiovascular disease. Mechanistic investigations have focused on intrarenal cellular signaling induced by ischemia/reperfusion. Additional cardiorenal connector signals have been postulated, but investigation in CRS-1 has been limited by a paucity of animal models and technical limitations precluding discovery studies of glomerular filtrate. To address these limitations, we developed a translational model of CRS-1, cardiac arrest and cardiopulmonary resuscitation (CA/CPR) in mice. Quantitative proteomics using isobaric tagging (10-plex TMT reagents, Thermo Fisher) was performed on 24h urine collections from mice with deficient tubular endocytosis and from littermate controls, before and after CA/CPR. Data acquisition used the SPS MS3 method on a Thermo Fusion Tribrid mass spectrometer for accurate reporter ion signals. The findings confirmed CA/CPR-specific cardiac proteins in urine and identified a novel CA/CPR-specific filtrate component: Cardiac LIM protein (CSRP3).
2019-02-27 | PXD012193 | Pride
Project description:Despite numerous studies reporting deregulated microRNA (miRNA) and gene expression patterns in clear cell renal cell carcinoma (ccRCC), no direct comparisons have been made to its presumed normal counterpart; the renal proximal epithelial tubular cells (PTEC). The aim of this study was to determine the miRNA expression profiles of ten clear cell renal cell carcinoma-derived cell lines and short-term cultures of PTEC, and to correlate these with their gene expression, and copy-number profiles. Using microarray-based methods, a significantly altered expression level in ccRCC cell lines was observed for 23 miRNAs and 1630 genes. The set of miRNAs with significantly decreased expression levels include all members of the miR-200 family known to be involved in the epithelial to mesenchymal transition (EMT) process. Expression levels of 13 of the 47 validated target genes for the downregulated miRNAs were increased more than two-fold. Our data reinforce the importance of the EMT process in the development of ccRCC. For mRNA expression data of these cell lines see GEO Series accession number GSE20491. MicroRNA profiling was performed on two proximal tubular epithelial cell samples (both cell samples were hybridized twice (biological duplicates)) and ten clear cell renal cell carcinoma- derived cell lines (one of which; RCC-JF in duplicate)
2012-11-01 | E-GEOD-34981 | biostudies-arrayexpress
Project description:O-GlcNAcylation is a global post translational modification. The purpose of this study is to investigate the role of O-GlcNAcylation in mice kidney proximal tubules. We used drug-inducible proximal tubular cells specific O-GlcNAc transferase (Ogt) KO mice (PTEC-Ogt KO). In this proteomic analysis, we compared kidney cortex from fasted PTEC-Ogt KO and Control mice.
2019-05-17 | PXD012456 | JPOST Repository
Project description:The endocytic receptor megalin constitutes the main pathway for clearance of plasma proteins from the glomerular filtrate in the proximal tubules. However, little is know about the mechanisms that control receptor activity. A widely discussed hypothesis states that the intracellular domain (ICD) of megalin, released upon ligand binding, acts as a transcription regulator to suppress receptor expression - a mechanism proposed to safeguard the proximal tubules from protein overload. Here, we have put this hypothesis to the test by generating a mouse model co-expressing the soluble ICD and the full-length receptor. Despite pronounced expression in the proximal tubules, the ICD failed to exert any effects on renal proximal tubular function such as megalin expression, protein retrieval, or renal gene transcription. Thus, our data argue that the ICD does not play a role in regulation of megalin activity in vivo in the proximal tubules. We used microarrays to compare gene expression profile in adult kidney from a new mouse model expressing the intracellular domain of megalin with wildtype. 10 week old mice were collected for RNA extraction and hybridization on Affymetrix microarrays. Three individuals for each genotype were analyzed comparing heterozygous animals for the intracellular domain of megalin with littermates controls.
2011-01-01 | E-GEOD-19778 | biostudies-arrayexpress
Project description:The microgravity environment aboard the International Space Station (ISS) provides a unique stressor that can help understand underlying cellular and molecular drivers of pathological changes observed in astronauts with the ultimate goals of developing strategies to enable longterm spaceflight and better treatment of diseases on Earth. We used this unique environment to evaluate the effects of microgravity on kidney proximal tubule epithelial cell (PTEC) response to serum exposure and vitamin D biotransformation capacity. To test if microgravity alters the pathologic response of the proximal tubule to serum exposure, we treated PTECs cultured in a microphysiological system (PT-MPS) with human serum and measured biomarkers of toxicity and inflammation (KIM-1 and IL-6) and conducted global transcriptomics via RNAseq on cells undergoing flight (microgravity) and respective controls (ground). We also treated 3D cultured PTECs with 25(OH)D3 (vitamin D) and monitored vitamin D metabolite formation, conducted global transcriptomics via RNAseq, and evaluated transcript expression of CYP27B1, CYP24A1, or CYP3A5 in PTECs undergoing flight (microgravity) and respective ground controls. We demonstrated that microgravity neither altered PTEC metabolism of vitamin D nor did it induce a unique response of PTECs to human serum, suggesting that these fundamental biochemical pathways in the kidney proximal tubule are not significantly altered by short-term 3 exposure to microgravity. Given the prospect of extended spaceflight, more study is needed to determine if these responses are consistent with extended (>6 month) exposure to microgravity.
2025-05-12 | GSE268236 | GEO
Project description:Effect of Cardiac LIM Protein (CSRP3) on human proximal tubule epithelial cells
| PRJNA1156043 | ENA
Project description:The endocytic receptor megalin constitutes the main pathway for clearance of plasma proteins from the glomerular filtrate in the proximal tubules. However, little is know about the mechanisms that control receptor activity. A widely discussed hypothesis states that the intracellular domain (ICD) of megalin, released upon ligand binding, acts as a transcription regulator to suppress receptor expression - a mechanism proposed to safeguard the proximal tubules from protein overload. Here, we have put this hypothesis to the test by generating a mouse model co-expressing the soluble ICD and the full-length receptor. Despite pronounced expression in the proximal tubules, the ICD failed to exert any effects on renal proximal tubular function such as megalin expression, protein retrieval, or renal gene transcription. Thus, our data argue that the ICD does not play a role in regulation of megalin activity in vivo in the proximal tubules. We used microarrays to compare gene expression profile in adult kidney from a new mouse model expressing the intracellular domain of megalin with wildtype.
2011-01-01 | GSE19778 | GEO