Unknown,Transcriptomics,Genomics,Proteomics

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Microarray analysis of human pediatric omental biopsies obtained from uremic patients, patients on peritoneal dialysis or healthy controls

ABSTRACT: Cardiovascular disease (CVD) is exceedingly severe in patients with chronic kidney disease (CKD) and further aggravated by peritoneal dialysis (PD), exposing the patients to excessive amounts of intraperitoneal glucose. Children are devoid of pre-existing CVD and give insight into specific uremia and PD induced pathomechanisms. Fat surrounded omental arterioles beyond PD fluid penetration level were microdissected from uremic children at time of first PD catheter insertion (n=8), children on PD (n=5), and age and gender matched non-uremic children as controls (n=6). Adjacent sections of 4 arterioles per patient were used for transcriptomic analyses.

ORGANISM(S): Homo sapiens

SUBMITTER: Claus Schmitt

PROVIDER: E-MTAB-5695 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Expression data from peritoneal biopsies of patients with encapsulating peritoneal sclerosis (EPS), patients undergoing first implantation of a peritoneal dialysis catheter (PD), and patients undergoing abdominal surgery for non-peritoneal conditions (controls) We used microarrays to determine the transcriptional profiles of peritoneal membrane in patients with encapsulating peritoneal sclerosis (EPS), patients undergoing first insertion of a peritoneal dialysis cathetier (PD), and uremic patients without history of PD or EPS, undergoing abdominal surgery for non-peritoneal problems (CON) Encapsulating peritoneal sclerosis (EPS) is a devastating complication of peritoneal dialysis (PD), characterized by marked inflammation and severe fibrosis of the peritoneum, and associated with high morbidity and mortality. EPS can occur years after termination of PD and, in severe cases, leads to intestinal obstruction and ileus requiring surgical intervention. Despite ongoing research, the pathogenesis of EPS remains unclear. We performed a global transcriptome analysis of peritoneal tissue specimens from EPS patients, PD patients without EPS, and uremic patients without history of PD or EPS (Uremic). Unsupervised and supervised bioinformatics analysis revealed distinct transcriptional patterns that discriminated these three clinical groups. The analysis identified a signature of 219 genes expressed differentially in EPS as compared to PD and Uremic groups. Canonical pathway analysis of differentially expressed genes showed enrichment in several pathways, including antigen presentation, dendritic cell maturation, B cell development, chemokine signaling and humoral and cellular immunity (P value <0.05). Further interactive network analysis depicted effects of EPS-associated genes on networks linked to inflammation, immunological response, and cell proliferation. Gene expression changes were confirmed by qRT-PCR for a subset of the differentially expressed genes. EPS patient tissues exhibited elevated expression of genes encoding sulfatase1, thrombospondin 1, fibronectin 1 and alpha smooth muscle actin, among many others, while in EPS and PD tissues mRNAs encoding leptin and retinol-binding protein 4 were markedly down-regulated, compared to Uremic group patients. Immunolocalization of Collagen 1 alpha 1 revealed that Col1a1 protein was predominantly expressed in the submesothelial compact zone of EPS patient peritoneal samples, whereas PD patient peritoneal samples exhibited homogenous Col1a1 staining throughout the tissue samples. The results are compatible with the hypothesis that encapsulating peritoneal sclerosis is a distinct pathological process from the simple peritoneal fibrosis that accompanies all PD treatment. Total RNA was isolated from frozen peritoneal biopsy specimens obtained at time of surgery. RNA was hybridized to Affymetrix arrays, and analyzed. Select transcripts were subjected to validation by rt-pcr and by immunodetection.

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Microarray analysis of human pediatric omental biopsies obtained from uremic patients, patients on peritoneal dialysis or healthy controls

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