Abstract
Recent research suggests that cytoprotective responses, such as expression of heat-shock proteins, might be inadequately induced in mesothelial cells by heat-sterilized peritoneal dialysis (PD) fluids. This study compares transcriptome data and multiple protein expression profiles for providing new insight into regulatory mechanisms. Two-dimensional difference gel electrophoresis (2D-DIGE) based proteomics and topic defined gene expression microarray-based transcriptomics techniques were used to evaluate stress responses in human omental peritoneal mesothelial cells in response to heat- or filter-sterilized PD fluids. Data from selected heat-shock proteins were validated by 2D western-blot analysis. Comparison of proteomics and transcriptomics data discriminated differentially regulated protein abundance into groups depending on correlating or noncorrelating transcripts. Inadequate abundance of several heat-shock proteins following exposure to heat-sterilized PD fluids is not reflected on the mRNA level indicating interference beyond transcriptional regulation. For the first time, this study describes evidence for posttranscriptional inadequacy of heat-shock protein expression by heat-sterilized PD fluids as a novel cytotoxic property. Cross-omics technologies introduce a novel way of understanding PDF bioincompatibility and searching for new interventions to reestablish adequate cytoprotective responses.
Highlights
Peritoneal dialysis (PD) is a cost effective and safe form of renal replacement therapy in end stage renal disease
For investigating potentially involved regulatory mechanisms both protein and RNA levels of Human peritoneal mesothelial cells (HPMC) undergoing treatment with heat-sterilized PDF (H-PDF) or F-PDF for 24 h were analyzed by 2D-DIGE and topic defined gene expression microarrays
These identifications were screened for overlaps with the transcripts successfully quantified in the mRNA microarray
Summary
Peritoneal dialysis (PD) is a cost effective and safe form of renal replacement therapy in end stage renal disease. In experimental PD we and others have shown that acute exposure to cytotoxic contents of PDF results in rapid induction of heat shock proteins (HSP) in mesothelial cells during the recovery phase, counteracting toxic injury [3,4,5,6]. We have described unexpectedly low HSP expression upon more extended exposure to diluted heatsterilized PDF [7]. Albeit this setting still represents a highly artificial system, the exposure to diluted cytotoxic properties of PDF likely reflects intraperitoneal conditions during a PD dwell more closely than acute exposure to pure PDF [8,9,10]. Heat-sterilization and storage of glucose-based PDF result in formation of highly reactive glucose degradation
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