PXD034985-1

PXD034985 is an original dataset announced via ProteomeXchange.

Title	Endothelial pathomechanisms in response to in vito exposure to differentz peritoneal dialysis fluids
Description	Loss of ultrafiltration capacity in peritoneal dialysis (PD) patients is often associated with peritoneal vascular changes, manifest as diabetes-like vasculopathy and angiogenesis. The endothelial monolayer lining the vessel lumen controls vessel barrier function and thereby influences peritoneal substrate transport and ultrafiltration. In this study, we investigated the influence of exposure of human umbilical vein endothelial cells (HUVEC) to different PD fluids. HUVECs were exposed to experimental solutions for up to 24 h. All test fluids were sterile-filtered before usage. Each experiment consisted of three independent samples in biological replicates on separate culture plates. For PD fluid incubation, cells were first exposed to pure PD fluids (Dianeal PD4 3.86% glucose, Physioneal 3.86% glucose, Extraneal, all Baxter, Castlebar, Ireland), or to a lab-made GDP-free PD fluid or to normal medium without growth factors as control. Cells were subsequently exposed for 24 h to the above solutions diluted 1:1 with culture medium and brought to 2% FCS. Cells were then washed three times (250 mM sucrose, 10 mM Tris/HCl, pH 7) and lysed in 125 µL per well of lysis buffer (30 mM Tris, pH 8.5, 7 M urea, 2 M thiourea, 4% CHAPS, 1 mM EDTA, one tablet of Complete Protease Inhibitor (Roche, Basel; Switzerland) per 100 ml, and one tablet of PhosStop Protease Inhibitor (Roche) per 100 mL). Total protein concentration was determined with the Pierce 660 Kit (Thermo) per manufacturer’s manual. Lysates were stored at -80°C until further processing. We investigated the molecular mechanisms of endothelial cell pathology during in vitro PD fluid exposure by proteomic and bioinformatic analysis of the endothelial cell response to the PD fluid induced stress, integrating current understanding of PD-related cellular injury and stress responses (Bender et al., Nephrol Dial Transplant, 2011, doi:10.1093/ndt/gfq484; Kratochwill et al., BioMed research international, 2015, doi:10.1155/2015/628158; Kratochwill et al., J Proteome Res, 2009, doi:10.1021/pr800916s; Lechner et al., J Proteome Res, 2010, doi:10.1021/pr9011574). We then evaluated the effect of AlaGln addition to conventional PD fluid on these parameters in endothelial cells. This strategy allowed characterization of endothelial cell injury and stress responses during exposure to different PD fluid.
HostingRepository	PRIDE
AnnounceDate	2022-08-12
AnnouncementXML	Submission_2022-08-11_16:04:22.742.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Klaus Kratochwill
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-06-30 02:55:33	ID requested
⏵ 1	2022-08-11 16:04:23	announced

Sacnun JM, Hoogenboom R, Eibensteiner F, Sobieszek IJ, Unterwurzacher M, Wagner A, Herzog R, Kratochwill K, Proteome-Wide Differential Effects of Peritoneal Dialysis Fluid Properties in an In Vitro Human Endothelial Cell Model. Int J Mol Sci, 23(14):(2022) [pubmed]

submitter keyword: Human umbilical vein endothelial cells, TMT, dialysis

Dr Klaus Kratochwill
contact affiliation	Medical University of Vienna
contact email	klaus.kratochwill@meduniwien.ac.at
lab head
Klaus Kratochwill
contact affiliation	Medical University of Vienna
contact email	klaus.kratochwill@meduniwien.ac.at
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD034985
     1. Label: PRIDE project
     2. Name: Endothelial pathomechanisms in response to in vito exposure to differentz peritoneal dialysis fluids