The effect of cigarette smoking on bronchoalveolar lavage protein profiles from patients with different interstitial lung diseases.

The proteomic approach applied to the analysis of BAL gives a panorama of the complex network of proteins of different origin and function and their modifications at alveolar level. Cigarette smoking may influence BAL protein composition and it represents the most relevant risk factor for several lung diseases. This review, for the first time, discusses the available literature regarding the effects of cigarette smoking on BAL protein composition of healthy subjects and patients affected by interstitial lung diseases (ILD). The comparison of BAL protein profiles of smokers and non-smoker healthy controls revealed alterations of proteins related to oxidative stress and protease/antiprotease imbalance (such as alpha 1 antitrypsin, alpha-1-antichymotrypsin, apolipoprotein A1, peroxiredoxin 1 and glutathione S transferase P). Smoking exposure leads to a significant dysregulation of a large number of molecular pathways involved in interstitial lung diseases and the proteomic studies applied to the study of BAL of idiopathic pulmonary fibrosis, sarcoidosis and other ILD contributed to clarify the underlying pathogenetic mechanisms facilitating ILD development and biomarker discovery.

Alveolar nitric oxide is related to periostin levels in idiopathic pulmonary fibrosis.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive diffuse lung disease leading to chronic respiratory failure and death in 3-5 years. Among potential prognostic biomarkers, alveolar nitric oxide (CaNO) and serum periostin showed to predict mortality and disease progression in these patients. The aim of this study is to investigate potential correlations between CaNO and serum periostin and evaluate their prognostic value. METHODS: Fifty-nine patients with IPF (47 males, 65.5±9.5 years old) were recruited in Siena Regional Referral Center for Interstitial Lung Disease. In this population, we retrospectively collected multiple-flows exhaled nitric oxide parameters and serum periostin at diagnosis and compared these values with a control group of 60 and 8 healthy volunteers, respectively. Clinical, functional and survival data were collected according to our Center follow-up program. RESULTS: IPF patients reported higher levels of CaNO but not of periostin in respect with healthy controls (P<0.0001 and P=0.1096, respectively). CaNO significantly correlated with periostin levels and TLCO% (P<0.0001 and P=0.0205, respectively). Patients with CaNO>6 ppb showed a worse prognosis, close to statistical significance (P=0.0628). No difference in survival time was found according to periostin levels. CONCLUSIONS: CaNO was significantly higher in IPF patients and was related to functional severity of disease. CaNO levels correlated with periostin, suggesting a potential common pathway between the biomarkers.

A system biology study of BALF from patients affected by idiopathic pulmonary fibrosis (IPF) and healthy controls.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disease characterized by progressive loss of the alveolar integrity, recruitment, and activation of myofibroblast, and excessive collagen deposition that resulted in loss of parenchymal architecture and lung function. Although etiology is unknown, major risk factor of disease development is represented by cigarette smoke or exposure to dust. AIMS: Aim of this proteomic study was to compare broncho alveolar lavage fluid protein profiles of IPF patients, never-smoker healthy control (nonsmoker control) and smoker control subjects in order to investigate proteins potentially related to disease progression and pathogenesis. METHODS: Broncho alveolar lavage fluid samples were resolved using 2D-PAGE and the differentially expressed proteins were identified by MS. The performed PCA statistically proved the correlation existing between differentially expressed spots in the three groups. Functional analysis of the identified proteins was performed by pathway and enrichment analysis by MetaCore and Database for Annotation, Visualization and Integrated Discovery. RESULTS: Interestingly, transcriptional factors NF-kB, PPARγ, and c-myc emerged as well as a group of functional hubs. Enrichment analysis suggested that Gene Ontology (GO) process networks and pathway maps involved in IPF included angiotensin system maturation, renin-angiotensin-aldosteron system, heme metabolism, coagulation system, response to hypoxia, oxidative stress, and iron transport. CONCLUSION: In conclusion, the combination of proteomic data with system biology platforms allowed us to amplify the information obtained processing the results and indicated the principal pathways involved. These information can point to potential biomarkers and new therapeutic targets opening the way for further analysis.

Calgranulin B (S100A9/MRP14): a key molecule in idiopathic pulmonary fibrosis?

Calgranulin B is a small calcium-binding protein with several immunological functions mainly involved in chronic inflammation and cancer. It can participate in recruitment of neutrophils and leukocytes in inflamed tissue, oxidant/antioxidant balance, adhesion of neutrophils to fibronectin, and regulation of apoptosis. In a previous proteomic study, we found that calgranulin B was up-regulated in the bronchoalveolar lavage (BAL) of patients with idiopathic pulmonary fibrosis (IPF) with respect to controls and patients with other interstitial lung diseases. The aims of this study are to compare calgranulin B concentrations in BAL of patients with IPF and sarcoidosis and controls by a quantitative method, to look for correlations with clinical data, and to evaluate calgranulin B expression in lung tissue of IPF patients by immunohistochemistry. A modification of a commercial ELISA was used to determine calgranulin B concentrations in BAL of 16 patients with IPF (a group of patients in which we previously performed proteomic analysis), 17 patients with sarcoidosis, and 7 controls. The immunohistochemistry was done in a subgroup of patients with IPF and a control group of lung transplant donors. Calgranulin B concentrations were significantly higher in patients with IPF than controls (p < 0.01); they were inversely correlated with FVC and DLCO values and directly correlated with neutrophil and eosinophil percentages in BAL. Immunohistochemistry revealed a patchy distribution of calgranulin B, predominantly around areas of fibrotic remodeling. Calgranulin B may be a trigger molecule involved in the evolution and progression of IPF, being overexpressed in BAL of patients with IPF with severe functional deterioration and in the peribronchiolar area bordering zones of honeycombing.