Dataset Information

CF sputum LC-MSMS - Proteomic profile of cystic fibrosis sputum cells in adults chronically infected with Pseudomonas aeruginosa

ABSTRACT: Lung disease is the main cause of morbidity and mortality in cystic fibrosis (CF), and involves chronic infection by a destructive microbiota and perturbed innate and adaptive immune responses. Tissue damage is considered to be mediated mostly by proteases, but other bacterial and host factors may also play a role. To determine the presence of potentially injurious proteins we employed semi-quantitative Multidimensional Protein Identification Technology to identify sputum cellular proteins with consistently altered expression in CF compared to healthy controls. Ingenuity Pathway Analysis, Gene Ontology functions, protein abundance and correlation with lung function were used to infer their clinical significance. The CF proteome exhibited differential expression of proteins relating to Rho family small GTPase activity, immune cell movement and activation, generation of reactive oxygen species and dysregulation of cell death and proliferation. Compositional breakdown established neutrophil extracellular trap proteins as the consistently most abundant cellular proteins detected, while a further 13 biologically relevant proteins were found to correlate negatively with lung function. These findings expand the current understanding of the mechanisms underlying CF lung disease and identify sputum cell proteins which might be useful as markers of disease status, prognostic indicators, stratification determinants for treatment prescription or as therapeutic targets.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Erythrocyte, Platelet, B Cell, T Cell, Leukocyte, Dendritic Cell, Sputum, Epithelial Cell, Neutrophil, Fibroblast, Macrophage

DISEASE(S): Cystic Fibrosis

SUBMITTER: S Pattison

LAB HEAD: Joseph Stuart Elborn

PROVIDER: PXD001985 | Pride | 2017-05-19

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	CF1+CF2(UniProt).pride.mztab.gz	Mztab
	CF1+CF2.pride.mztab.gz	Mztab
	CF12+CF13(UniProt).pride.mztab.gz	Mztab
	CF12+CF13.pride.mztab.gz	Mztab
	CF12CF13.mzid.gz	Mzid

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Proteomic profile of cystic fibrosis sputum cells in adults chronically infected with <i>Pseudomonas aeruginosa</i>.

Pattison Sally H SH Gibson David S DS Johnston Elinor E Peacock Samantha S Rivera Keith K Tunney Michael M MM Pappin Darryl J DJ Elborn J Stuart JS

The European respiratory journal 20170705 1

Lung disease is the main cause of morbidity and mortality in cystic fibrosis (CF), and involves chronic infection and perturbed immune responses. Tissue damage is mediated mostly by extracellular proteases, but other cellular proteins may also contribute to damage through their effect on cell activities and/or release into sputum fluid by means of active secretion or cell death.We employed MudPIT (multidimensional protein identification technology) to identify sputum cellular proteins with consi ...[more]

PMID: 28679606

Similar Datasets

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Project description:One of the hallmarks of Pseudomonas aeruginosa cystic fibrosis (CF) infection is very high-cell-density (HCD) replication in the lung, allowing this bacterium to induce virulence controlled by HCD quorum-sensing systems. However, the nutrient sources sustaining HCD replication in this chronic infection is largely unknown. Hence, understanding the nutrient factors contributing to HCD in the CF lung will yield new insights into the 'metabolic pathogenicity' and potential treatment of CF infections caused by P. aeruginosa. Herein, we performed microarray studies of P. aeruginosa directly isolated from the CF lung to demonstrate its metabolic capability and virulence in vivo. Our in vivo microarray data, confirmed by real-time reverse-transcription-PCR, indicated P. aeruginosa expressed several genes for virulence, drug-resistance, and utilization of multiple nutrient sources (lung surfactant lipids and amino acids) contributing to HCD replication. The data also indicates deregulation of several pathways, suggesting in vivo evolution by deregulation of a large portion of the transcriptome during chronic CF infection. To our knowledge, this is the first in vivo transcriptome of P. aeruginosa in a natural CF infection, and it indicates several important aspects of pathogenesis, drug-resistance, and nutrient-utilization never before observed in vivo. Experiment Overall Design: The purpose of the experiment was to observe which genes are upregulated in P. aeruginosa during chronic CF lung infection as compared to PAO1. All in vitro studies were grown in 1x M9 minimal media supplemented with 20 mM citrate and grown to mid-log phase prior to RNA isolation. The in vivo RNA was isolated directly from CF sputum samples after TRIzol treatment. Each in vitro sample (both for PAO1 and the CF sputum pool isolate) were processed individually and in triplicate. Two in vivo isolations from sputum were conducted from the same patient but two different sputum samples. After isolation of total RNA, samples were processed for microarrays (i.e. cDNA synthesis, fragmentation, labeling, etc) as recommended by Affymetrix, and processed on the GeneChip as recommended by Affymetrix.

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