Proteomic Fingerprint of Lung Fibrosis Progression and Response to Therapy in Bleomycin-Induced Mouse Model

Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by the aberrant accumulation of extracellular matrix in the lungs. nintedanib is one of the two FDA-approved drugs for IPF treatment; however, the exact pathophysiological mechanisms of fibrosis progression and response to therapy are still poorly understood. In this work, the molecular fingerprint of fibrosis progression and response to nintedanib treatment have been investigated by mass spectrometry-based bottom-up proteomics in paraffin-embedded lung tissues from bleomycin-induced (BLM) pulmonary fibrosis mice. Our proteomics results unveiled that (i) samples clustered depending on the tissue fibrotic grade (mild, moderate, and severe) and not on the time course after BLM treatment; (ii) the dysregulation of different pathways involved in fibrosis progression such as the complement coagulation cascades, advanced glycation end products (AGEs) and their receptors (RAGEs) signaling, the extracellular matrix-receptor interaction, the regulation of actin cytoskeleton, and ribosomes; (iii) Coronin 1A (Coro1a) as the protein with the highest correlation when evaluating the progression of fibrosis, with an increased expression from mild to severe fibrosis; and (iv) a total of 10 differentially expressed proteins (padj-value ≤ 0.05 and Fold change ≤−1.5 or ≥1.5), whose abundance varied in the base of the severity of fibrosis (mild and moderate), were modulated by the antifibrotic treatment with nintedanib, reverting their trend. Notably, nintedanib significantly restored lactate dehydrogenase B (Ldhb) expression but not lactate dehydrogenase A (Ldha). Notwithstanding the need for further investigations to validate the roles of both Coro1a and Ldhb, our findings provide an extensive proteomic characterization with a strong relationship with histomorphometric measurements. These results unveil some biological processes in pulmonary fibrosis and drug-mediated fibrosis therapy.

Innovative Pre-Clinical Data Using Peptides to Intervene in the Evolution of Pulmonary Fibrosis

“…The major IPF experimental model is bleomycin-treated C57bl6 mice [ 12 , 25 , 26 ]. However, these animals develop acute fibrosis, which leads to self-healing 21 days post-instillation (p.i.)…”

Proteomic analysis of transbronchial biopsy tissue reveals a distinct proteome and mechanistic pathways in high-grade eosinophilic inflammation after lung transplantation

“…CCT8, a subunit of the cytoplasmic chaperonin containing TCP1 complex (CCT), is necessary for correct folding of many proteins. Recent experimental studies have shown that overexpression of CCT8 not only promotes EMT, but also involved in lung brosis development and progression (34,40). Thus, up-regulation of CCT8 in the TBBx of LTx patients with EOS and ACR may be an important clinical indicator for the development of brosis and graft rejection in these cases.…”

“…Coro1A, also known as translational activator of cytochrome oxidase, regulates various cellular processes such as migration and endocytosis. Coro1A overexpression is signi cantly correlated with in ammatory processes, oxidative stress and the progression of lung brosis (40,41). Recent evidence has revealed higher expression of Coro1A in brotic tissues than normal cells, and that Coro1A expression level was directly related to brosis severity (42).…”

Feasibility of MALDI-MSI-Based Proteomics Using Bouin-Fixed Pathology Samples: Untapping the Goldmine of Nephropathology Archives

“…To perform protein identification and quantification, a pool of 6 slices of BFPE and FFPE renal samples at three different fixation time points (2, 12, and 24 h), along with a pool of 5 slices of BFPE and FFPE renal amyloidosis, were collected and prepared for nLC-ESI MS/MS analysis. Samples were deparaffinized and digested as already described . After the paraffin removal, antigen retrieval, and trypsinization, the enzymatic reaction was stopped by adding TFA to a final concentration of 0.5% to reach an acidic pH (<2).…”