P311 KO Mice are Protected from Experimentally Induced Lung Fibrosis

Abstract

Pulmonary fibrosis (PF) is a fatal chronic interstitial lung disease and one of the most irreversible forms of tissue/organ fibrosis. Idiopathic pulmonary fibrosis is a very progressive disease accompanied by inflammatory damage with a median survival of less than 5 years. Etiology of IPF is still elusive with potential reasons being genetic predisposition and environmental exposures including particulates, radiation, and SARS-CoV-2. Recent studies indicate that about 50% of COVID19 associated acute respiratory distress syndrome survivors develop PF. There are very limited treatment options with no curative therapies available for PF apart from invasive lung transplantation. P311 is an 8 kDa protein previously shown to have a key role in lung regeneration. As P311 overexpression in fibroblasts induced myofibroblast phenotype, we hypothesized that P311 has a role in tissue fibrosis. We induced PF by intra-tracheally instilling bleomycin (BL) in P311 knockout (KO) mice and wild type (WT, C57BL/6) control mice and collected lungs after 2-5 weeks. P311 KO mice showed significantly less fibrosis. After intra-tracheal instillation of BL, the lungs of P311 KO mice (D21) had significantly reduced number of myofibroblasts as indicated by reduced SM α-actin expression and collagen deposition as indicated by Masson trichrome staining compared to their WT counterparts. We further observed reduced levels of TGF-β1-3 and Smad-2 and -3 along with decreased epithelial to mesenchymal transdifferentiation markers in the lungs of bleomycin treated P311 KOs compared to WT controls. Further bronchoalveolar lavage fluid (BALF) was collected (D14) and utilized to determine the pulmonary inflammation by determining the total and different immune cell populations in the BALF and lung samples by FACS analysis. Even though we saw a significant increase in the BALF cellularity, we did not observe significant changes in T-cell, macrophage, and neutrophil numbers between WT and P311KOs. However, lung cell isolates showed significantly increased CD3 and CD4 positive cells indicating increased T-cells and helper T-cells in fibrotic P311KO lungs compared to WT lungs. In conclusion, bleomycin administration resulted in lung fibrosis in WT mice, but P311 KO mice are protected. This study was partially supported by research funding to KB, from American Lung Association, DA-196629-N. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.