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 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 consistently altered abundance in adults with CF, chronically infected with , compared with healthy controls. Ingenuity Pathway Analysis, Gene Ontology, protein abundance and correlation with lung function were used to infer their potential clinical significance.Differentially abundant proteins relate to Rho family small GTPase activity, immune cell movement/activation, generation of reactive oxygen species, and dysregulation of cell death and proliferation. Compositional breakdown identified high abundance of proteins previously associated with neutrophil extracellular traps. Furthermore, negative correlations with lung function were detected for 17 proteins, many of which have previously been associated with lung injury.These findings expand our current understanding of the mechanisms driving CF lung disease and identify sputum cellular proteins with potential for use as indicators of disease status/prognosis, stratification determinants for treatment prescription or therapeutic targets.

The microbiome in bronchiectasis

“…Disease-associated exacerbations have a major effect on patient healthcare costs as well as quality of life due to increased lung damage and mortality risk [10][11][12][13]. Microorganisms such as Pseudomonas aeruginosa and, to a lesser extent, other Gram-negative and Gram-positive microorganisms identified in culture, have been linked to disease progression, poor clinical outcomes in bronchiectasis and driving airway neutrophil-mediated inflammation [14][15][16][17][18][19]. For this reason, long-term antibiotic therapy is one of the mainstays of treatment for the management of bronchiectasis with frequent exacerbations [10,13,20,21].…”

Serum-Based Proteomics Profiling in Adult Patients with Cystic Fibrosis

“…Proteases are involved in a series of intracellular and extracellular regulatory processes, including tissue remodeling, mucin expression, bacterial killing, and neutrophil chemotaxis. In the healthy lung, proteases and antiproteases maintain a homeostatic balance, preventing lung damage that may arise from proteases [ 6 , 18 ]. However, the CF lung is a protease-rich environment, and this protease burden overwhelms the antiprotease capabilities, leading to a protease-antiprotease imbalance, which is heavily implicated in CF pathophysiology [ 6 , 12 ].…”

“…However, discovering new biomarkers to evaluate disease progression and therapeutic targets to reduce CF complications is still necessary. Proteomics has evolved as a promising platform for the identification of secreted proteins involved in CF pathophysiology, which could help in discovering new biomarkers or therapeutic targets for CF [ 2 , 12 , 18 ].…”

“…In primary cultures of human nasal epithelial cells obtained from CF patients with nasal polyps, proteomic analysis revealed changes in pathways related to metabolism, G-protein processing, inflammation, oxidative stress response, protein folding, proteolysis, and structural proteins [ 9 ]. An analysis of the sputum cellular proteins of CF patients with chronic Pseudomonas aeruginosa infection indicated the involvement of the Rho family of small GTPases, immune cell movement/activation, the generation of ROS, and the dysregulation of cell death and proliferation [ 18 ]. Another proteomic analysis of sputum from CF patients showed the differential expression of proteins related to proteolytic degradation and the influx of inflammation, such as myeloperoxidase, cleaved alpha-1-antitrypsin, IgG degradation, and IL8 [ 14 ].…”

Toll like Receptor signalling by Prevotella histicola activates alternative NF-κB signalling in Cystic Fibrosis bronchial epithelial cells compared to P. aeruginosa

“…Activation of the alternative pathway is known to occur in response to a small set of agonists including lipopolysaccharide (LPS), CD40Ligand and lymphotoxin-alpha (LT-α 1 ). This pathway has been shown to be active in epithelial cells and has recently been implicated in pathway analysis of inflammation in people with CF [ 15 – 17 ]. Inhibitor of kappaB kinase α (IKKα) plays a role in resolving NF-κB-driven inflammation in TLR expressing cells by facilitating turnover of p65 and cRel, removing them from the nucleus [ 18 ].…”