Proteome characterization of BALF extracellular vesicles in Idiopathic Pulmonary Fibrosis: unveiling undercover molecular pathways.

ABSTRACT: In the longtime challenge of identifying specific, easily-detectable and reliable biomarkers of interstitial lung diseases (ILDs), alternative biofluids, such as bronchoalveolar lavage fluid (BALF), have been extensively studied. In particular, BALF proteomics is gaining momentum and providing interesting new insights into the pathogenesis of ILDs, especially Idiopathic Pulmonary Fibrosis (IPF). Interest in the role of extracellular vesicles (EVs) in the pathogenesis of IPF has also grown. Though few, current studies on EVs in BALF of IPF patients focus mainly on miRNAs. To the best of our knowledge, the present study is therefore the first shotgun proteomic investigation of EVs isolated from BALF of IPF patients. Our main aim was to characterize the proteome of the vesicular component of BALF and to explore its individual impact on the pathogenesis of IPF. To this purpose, ultracentrifugation was chosen as EVs isolation technique and their purification was assessed by TEM, 2DE and LC-MS/MS. Our 2DE data and scatter plots showed considerable differences between the proteome of EVs and that of its supernatant and of whole BALF. Analysis of protein content and protein functions evidenced that EV proteins are predominantly involved in cytoskeleton remodeling, adenosine signaling, adrenergic signaling, C-peptide signaling and lipid metabolism. Our findings may suggest a wider system involvement in the disease pathogenesis and support the importance of pre-fractioning of complex samples, like BALF, in order to let low-abundant proteins-mediated pathways to emerge.

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Bronchoalveolar Lavage

DISEASE(S): Idiopathic Pulmonary Fibrosis

SUBMITTER: Enxhi Shaba  

LAB HEAD: Prof. Luca Bini

PROVIDER: PXD025590 | Pride | 2021-08-06

REPOSITORIES: Pride