PXD022183

PXD022183 is an original dataset announced via ProteomeXchange.

Title	Endothelial pathomechanisms in response to in vivo peritoneal dialysis fluid exposure
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 first characterized the response to conventional PD fluids of endothelial cells isolated from peritoneal biopsies of children undergoing PD. 9 omental biopsies from the pediatric biopsy biobank (mean age 5.7 years) were included in this study. PD patients were treated with conventional PD fluid (mean PD vintage 12.5 months). Age-matched controls (n=5) with normal renal function undergoing elective surgery were also included. In patients on PD, the biopsy sampling site was at least 5 cm away from the PD catheter entry site. Written informed consent was obtained from parents, and from patients as appropriate. The study was performed according to the Declaration of Helsinki and registered at www.clinicaltrials.gov (NCT01893710). The study was part of the International Pediatric Dialysis Network (IPDN; www.pedpd.org). Patients with a BMI of >35 kg/m2 and with chronic inflammatory diseases were excluded. Specimens obtained during surgery were instantaneously frozen with liquid nitrogen and stored at -80°C. Arterioles macroscopically located within fat tissue and microscopically at least 1mm distant from the peritoneal surface were micro-dissected. Elastica van Gieson stainings of the arteriolar elastic lamina were used as templates for microdissection of arterioles from cresol violet–stained neighboring sections. 100 µm thick tissue slices were cut with a cryotome and 30 deep-frozen arteriolar rings per patient were micro-dissected using a stereo microscope and a microneedle. Arteriolar rings were lysed in 100 µl lysis buffer (30 mM Tris, pH 8.5, 7M urea, 2M thiourea, 4% 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), 1 mM EDTA, 1 tablet of Complete Protease Inhibitor (Roche, Basel, Switzerland) and 1 tablet of phosphatase inhibitor (PhosSTOP, Roche) per 100 ml). The resulting lysates were stored at -80°C until further processing. Total protein concentration was determined with the 2D-Quant kit (GE). Quality control and verification of protein concentrations was performed by SDS-PAGE. We investigated the molecular mechanisms of endothelial cell pathology during in vivo and in vitro PD fluid exposure by proteomic and bioinformatic analysis of the endothelial cell response to hyperglycemic 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).
HostingRepository	PRIDE
AnnounceDate	2021-09-09
AnnouncementXML	Submission_2021-09-09_13:11:32.673.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	2020-10-26 03:07:51	ID requested
⏵ 1	2021-09-09 13:11:33	announced

Herzog R, Bartosova M, Tarantino S, Wagner A, Unterwurzacher M, Sacnun JM, Lichtenauer AM, Kuster L, Schaefer B, Alper SL, Aufricht C, Schmitt CP, Kratochwill K, Peritoneal Dialysis Fluid Supplementation with Alanyl-Glutamine Attenuates Conventional Dialysis Fluid-Mediated Endothelial Cell Injury by Restoring Perturbed Cytoprotective Responses. Biomolecules, 10(12):(2020) [pubmed]

submitter keyword: human arterioles, 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. PXD022183
     1. Label: PRIDE project
     2. Name: Endothelial pathomechanisms in response to in vivo peritoneal dialysis fluid exposure