Gene expression profile of human peritoneal mesothelial cells (HPMC) treated with TGF-b1
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE121372 | GEO | 2018/10/18
REPOSITORIES: GEO
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Project description:Peritoneal fibrosis is a major complication of long-term peritoneal dialysis (PD), leading to ultrafiltration failure and sometimes life threatening encapsulating peritoneal sclerosis. Fibrosis is driven by activated myofibroblasts that are derived, in part, from mesothelial-to-mesenchymal transition (MMT). We aimed to discover novel mediators of MMT and then experimentally exploit them to prevent peritoneal fibrosis. Using an antibody to HBME-1 and streptavidin nanobead technology, we first pioneered a novel method to purify rat mesothelial cells. After exposing mesothelial cells to transforming growth factor β1 (TGFβ1), we undertook RNAseq whole transcriptome analyses and outlined, the expression profile of sorted mesothelial cells at pre- and post- MMT.
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Project description:Peritoneal mesothelial cells are harmed by peritoneal dialysis fluids (PDF) used in renal replacement therapy with peritoneal dialysis. The mechanisms of the cellular damage are not yet described in detail. Primary human peritoneal mesothelial cells derived from omentum of five donors were independently exposed to peritoneal dialysis fluids (extended recovery time). The extent of cell damage was assessed using lactate dehydrogenase (LDH) release in the cell culture supernatant and cells were lysed in order to extract mRNA and proteins. Transcriptional changes induced by PDF were analyzed using gene expression microarrays and changes of the proteome were analyzed using 2D-electrophoresis.
2019-12-31 | E-MTAB-7793 | biostudies-arrayexpress
Project description:Peritoneal mesothelial cells are harmed by peritoneal dialysis fluids (PDF) used in renal replacement therapy with peritoneal dialysis. The mechanisms of the cellular damage are not yet described in detail. Primary human peritoneal mesothelial cells derived from omentum of five donors were independently exposed to peritoneal dialysis fluids. The extent of cell damage was assessed using lactate dehydrogenase (LDH) release in the cell culture supernatant and cells were lysed in order to extract mRNA and proteins. Transcriptional changes induced by PDF were analyzed using gene expression microarrays and changes of the proteome were analyzed using 2D-electrophoresis.
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Project description:Peritoneal dialysis (PD) is the worldwide recognized preferred dialysis treatment for children affected by end-stage kidney disease (ESKD). However, due to the unphysiological composition of PD fluids, the peritoneal membrane (PM) of these patients may undergo structural and functional alterations, which may cause fibrosis. Several factors may accelerate this process and primary kidney disease may have a causative role. In particular, patients affected by corticoresistant primary focal segmental glomerulosclerosis), a rare glomerular disease leading to nephrotic syndrome and ESKD, seem more prone to develop peritoneal fibrosis. The mechanism causing this predisposition is still unrecognized. To better define this condition, we carried out, for the first time, a new comprehensive comparative proteomic mass spectrometry analysis of mesothelial exosomes from peritoneal dialysis effluent (PDE) of 6 pediatric patients with focal segmental glomerular sclerosis (FSGS) versus 6 patients affected by other primary renal diseases (No FSGS). Our omic study demonstrated that, despite the high overlap in the protein milieu between the two study groups, machine learning allowed a complete distinction of the whole proteomic exosome mesothelial content of FSGS versus No FSGS (with 100% accuracy). Out of the 2490 identified proteins, 40% (995) were involved in epithelial-mesenchymal transition (EMT)/fibrosis and in the transforming growth factor-β pathway. Additionally, the Weight Gene Co-expression Network Analysis algorithm identified that some of the discriminative proteins (TIMP1, CTHRC1, SPARC, CHMP4B, COL5A2, ANXA13, FNC2 and CENP-E) were also highly correlated to PD vintage, fibrosis, EMT and non-neoplastic PM disease. All together our data demonstrated that mesothelial cells of FSGS patients are more prone to activate a pro-fibrotic machinery with exosomes having a primary role in this process. Moreover, they indicated that identified FSGS-associated elements in mesothelial exosome protein could be employed as potential new biomarkers of mesothelial integrity. Finally, our results highlighted that in FSGS patients particular attention should be paid to use more biocompatible dialysis solution, reduce the length of time on PD and personalize PD regimens to minimize the risk of rapid loss of PM function or development of encapsulating peritoneal sclerosis.
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Project description:Peritoneal mesothelial cells are harmed by peritoneal dialysis fluids (PDF) used in renal replacement therapy with peritoneal dialysis. The mechanisms of the cellular damage are not yet described in detail. Primary human peritoneal mesothelial cells derived from omentum of five donors were independently exposed to peritoneal dialysis fluids. The extent of cell damage was assessed using lactate dehydrogenase (LDH) release in the cell culture supernatant and cells were lysed in order to extract mRNA and proteins. Transcriptional changes induced by PDF were analyzed using gene expression microarrays and changes of the proteome were analyzed using 2D-electrophoresis.
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Project description:Background: Malignant ascites is often present at diagnostic in women with advanced ovarian cancer (OC) and its presence is associated with a worse outcome. Human peritoneal mesothelial cells (HPMCs) are key components of malignant ascites. Although the interplay between HPMCs and OC cells is believed to be critical for tumor progression, it has not been well characterized. The purpose of this study was to assess the effect of ascites on HPMCs and clarify the role of HPMCs in OC progression. Methods: Human OC ascites and benign peritoneal fluids were assessed for their ability to stimulate HPMC proliferation. Conditioned medium from ascites- and benign fluid-stimulated HPMCs were compared for their ability to attenuate apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL). We conducted a comparative analysis of global expression changes in ascites-stimulated HPMCs using Agilent oligonucleotide microarrays. Results: As compared to benign peritoneal fluids, malignant ascites stimulated the proliferation of HPMCs. TRAIL-induced apoptosis was attenuated in OC cells exposed to conditioned medium from ascites-stimulated HPMCs as compared to OC cells exposed to conditioned medium from benign fluid-stimulated HPMCs. A total of 649 genes were differentially expressed in ascites-stimulated HPMCs. Based on a ratio of more than 1.5-fold and a P < 0.05, 484 genes were up-regulated and 165 genes were down-regulated in ascites-exposed HPMCs. Stimulation of HPMCs with OC ascites resulted in differential expression of genes mainly associated with the regulation of cell growth and proliferation, cell death, cell cycle and cell assembly and organization, compared to benign peritoneal fluids. Top networks up-regulated by OC ascites included Akt and NF-M-NM-:B survival pathways whereas vascular endothelial growth factor (VEGF) pathway was down-regulated. Conclusions: The results of this study not only provide evidence supporting the importance of the interplay between cancer cells and HPMCs but also define the role that the tumor environment plays in these interactions. The expression profiles from human peritoneal mesothelial cells (HPMC) cultures exposed to peritoneal fluids and ovarian cancer ascites were compared using the Agilent Whole Human Genome Oligonucleotide microarrays, containing 44,000 genes. Microarrays were performed on HPMCs exposed to 3 malignant ascites from women with advanced (stage III/IV) serous OC and two benign peritoneal fluids. First, we generated lists of significantly up-regulated and down-regulated genes that were differentially expressed between OC ascites (OVC346, OVC508 and OVC509) and control OV370 peritoneal fluid. Then, the set of genes that were commonly expressed between control peritoneal fluids (OV370 and OV401) were subtracted from the first list of genes to generate a dataset of differentially expressed genes between malignant ascites and benign peritoneal fluids.
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Project description:Peritoneal fibrosis is regard as a significant cause of the loss of peritoneal function, markedly limiting the application of peritoneal dialysis(PD). However, the pathogenesis of peritoneal fibrosis remains unclear. Tissue-derived extracellular vesicles(EVs) mediate intercellular communications and play a central role in organ fibrosis. We performed a proteomics profiling of EVs from normal peritoneum and PD-induced fibrotic peritoneum in mice.
2023-01-25 | PXD037945 | Pride
Project description:Local factors produced in the tissue microenvironment play essential roles in promoting the ontogeny and phenotype of tissue resident macrophages (TRM). In the peritoneal cavity, large peritoneal macrophages (LPM) are the dominant TRMs that functionally mediate type 2 immunity, facilitate tissue repair of the mesothelium, and protect against peritoneal fibrosis. It is established that retinoic acid derived from the omentum induces transcription factor Gata6 expression in LPMs, which in turn regulates gene expression of factors that define peritoneal macrophages. It is still unclear whether retinoic acid is the sole local factor that regulates Gata6 expression in LPMs. Mesothelial cells line the entire peritoneal cavity and produce a protective, non-adhesive barrier against injury, at least in part by recruiting immune cells with secreted cytokines, such as M-CSF. We hypothesized that secreted factors from peritoneal mesothelial cells are also responsible for regulating LPM development including both ontogeny and function. Due to their immediate proximity to the peritoneal cavity, we propose that mesothelial cells can produce and secrete proteins into the peritoneum to maintain Gata6 expression by LPMs. To identify secreted factors that are highly and specifically expressed in mesothelial cells, we harvested primary mesothelial cells from 10-week-old C57BL/6 mice using FACS selection (CD45- PDPN+ GPM6a+). Total RNA was isolated from these cells and subjected to RNA-seq analysis after depletion of ribosomal RNA.
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Project description:Fushen Granule (FSG), a Chinese medicinal formular, was clinically used to improve the peritoneal dialysis (PD) efficiency in ESRD patients received PD treatment. However, its mechanisms of anti-fibrotic effect in peritoneal fibrosis (PF) has not been studied yet. In this work, we used TGF-β1-stimulated MeT5A cells to mimic the process of PF in vitro, and Label-free proteomics and bioinformatics analysis were used to explored the underlying targets and pathways of FSG on TGF-β1-treated MeT5A cells
2022-04-20 | PXD033058 | Pride
Project description:Gene expression profile of human peritoneal mesothelial cells (HPMC) treated with TGF-b1
| PRJNA497194 | ENA