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The Response of A549 Cells to Aspergillus fumigatus and Pseudomonas aeruginosa: A Proteomic perspective

ABSTRACT: Aspergillus fumigatus and Pseudomonas aeruginosa are the most prevalent fungal and bacterial pathogens, associated with cystic fibrosis-related infections, respectively. Although it is well established that P. aeruginosa eventually predominates as the primary pathogen, it is unknown why this is the case. Label-free quantitative proteomics was employed here to compare the cellular response to A. fumigatus, P. aeruginosa and sequential exposure to both pathogens in the type II alveolar epithelial cell line, A549, a model of the alveolar surface. In total, 2264 proteins were identified and the analysis of statistically significant (p<0.05; ±1.5 fold change), differentially abundant (SSDA) proteins, revealed an increase in the relative abundance of proteins associated with biological processes common to all pathogen-exposed groups. These included mitochondrial stress, energy metabolism and protein processing in the endoplasmic reticulum. Conversely, clear differences were observed between the cellular response to A. fumigatus or P. aeruginosa and following sequential exposure to both pathogens. The relative abundance of proteins associated with ribosomal activity appeared comparable between sequentially exposed cells and unexposed cells, but were substantially decreased in fungal- and bacterial-exposed cells. Furthermore, compared to unexposed cells, the relative abundance of proteins associated with the actin cytoskeleton, endocytosis, phagosome and lysosome were increased in A. fumigatus and P. aeruginosa-exposed cells, but appeared unchanged or decreased in sequentially exposed cells. This research provides novel insights into the whole cell proteomic response to A. fumigatus and P. aeruginosa, and highlights distinct differences following sequential exposure to both pathogens.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: James Carolan

LAB HEAD: James Carolan

PROVIDER: PXD014022 | Pride | 2019-11-11

REPOSITORIES: Pride

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Characterization of the Proteomic Response of A549 Cells Following Sequential Exposure to Aspergillus fumigatus and Pseudomonas aeruginosa.

Margalit Anatte A Kavanagh Kevin K Carolan James C JC

Journal of proteome research 20191114 1

Aspergillus fumigatus and Pseudomonas aeruginosa are the most prevalent fungal and bacterial pathogens associated with cystic-fibrosis-related infections, respectively. P. aeruginosa eventually predominates as the primary pathogen, though it is unknown why this is the case. Label-free quantitative proteomics was employed to investigate the cellular response of the alveolar epithelial cell line, A549, to coexposure of A. fumigatus and P. aeruginosa. These studie ...[more]

PMID: 31693381

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Project description:The airways of cystic fibrosis patients are chronically colonized by a diverse range of microbial pathogens, the composition of which changes throughout life Alteration to the pulmonary environment caused by inter-microbial interactions and pathogen-host interactions influence the type of microbes that can engage in sustained infection. The opportunistic bacterial pathogen Pseudomonas aeruginosa is the primary cause of morbidity and mortality amongst individuals with cystic fibrosis and it is estimated that 60 – 80 % of cystic fibrosis patients experience chronic P. aeruginosa infection by the age of 20 years Aspergillus fumigatus is the most prevalent fungal pathogen isolated from cystic fibrosis airways, affecting up to 58% of patients. It is the causative agent of allergic bronchopulmonary aspergillosis (ABPA), a hypersensitivity disorder resulting from the inhalation of fungal conidia. Although co-colonization of the cystic fibrosis airways by P. aeruginosa and A. fumigatus is rare (3.1 – 15.8 %) disease prognosis is poor when both are present. However, sequential infection is more common. It has recently been suggested that A. fumigatus is more prevalent in juvenile cystic fibrosis patients that has been initially reported due to inconsistencies in the culture methods used to detect A. fumigatus. Despite the prevalence and persistence of A. fumigatus, P. aeruginosa predominates as the primary pathogen in the cystic fibrosis lung, suggesting that interactions with other pathogens such as A. fumigatus may influence the pathogenicity of P. aeruginosa by altering its virulence. We report here an investigation of the effect of culturing P. aeruginosa in the presence of A. fumigatus by measuring differences in growth rate and the overall proteome of the bacteria. It was hypothesized that A. fumigatus creates an environment that promotes a metabolic-driven increase in P. aeruginosa that results in it outcompeting the fungus. The molecular basis of this increased proliferation was investigated further using Label-free quantitative (LFQ) proteomics to characterise the proteome changes in P. aeruginosa when exposed to the supernatant of i) A. fumigatus alone ii) the supernatant of an A. fumigatus/P. aeruginosa co-culture and iii) P. aeruginosa alone. LFQ proteomics involves the simultaneous identification and quantification of thousands of proteins (the ultimate determinants of phenotype) from a single sample has recently been employed to characterise the P. aeruginosa proteome in response to iron limiting conditions, resolving how how the bacteria survives and proliferates in such environments.

			Action	DRS
	Aspergillus_Rep_1.raw	Raw
	Aspergillus_Rep_2.raw	Raw
	Aspergillus_Rep_3.raw	Raw
	Control_Rep_1.raw	Raw
	Control_Rep_2.raw	Raw

Dataset Information

The Response of A549 Cells to Aspergillus fumigatus and Pseudomonas aeruginosa: A Proteomic perspective

Dataset's files

Publications

Characterization of the Proteomic Response of A549 Cells Following Sequential Exposure to <i>Aspergillus fumigatus</i> and <i>Pseudomonas aeruginosa</i>.

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