Inhibition of TGF-β/Smad signaling attenuates extracellular matrix accumulation in a human ex-vivo model of pulmonary fibrosis
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Date
2024-05
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The Ohio State University
Abstract
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive lung disease characterized by accumulation of scar tissue and decreased lung function. Mechanisms of disease onset have yet to be fully understood. However, activation of transforming growth factor-beta (TGF-β)/Smad signaling, followed by repeated injury to the epithelium, results in transdifferentiation of fibroblasts into profibrotic fibroblasts, leading to an increase in collagen expression and accumulation in the extracellular matrix (ECM), resulting in deteriorating lung function. Therefore, we proposed a TGF-β/Smad inhibitor to prevent ECM accumulation. We used human precision cut-lung slices (hPCLS) treated with TGF-β as a human ex-vivo model to retain native lung tissue structure, cellular complexity, and ECM protein composition. hPCLS were stimulated for 48 h with active recombinant human TGF-β (10ng/ml). Then, a TGF-β/Smad inhibitor (SB431542, 10µM) was added to the culture for 72 h and hPCLS were later harvested and analyzed to evaluate the expression of ECM proteins by immunofluorescence and PCR. We found high expression of collagen 1a1, collagen 3a1, fibronectin, and α-SMA after TGF-β stimulation. Treatment with the TGF-β/Smad inhibitor attenuated their expression. Our data suggests that the ex vivo model of hPCLS can be used to evaluate anti-fibrotic therapies. We demonstrated that specific inhibition of TGF- β results in the attenuation of the expression and deposition of proteins of the ECM. These results corroborate that modulation of TGF-β activity using SB431542 inhibitor has a therapeutic potential for patients with pulmonary fibrosis.
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Keywords
Extracellular matrix, pulmonary fibrosis, signaling inhibition, therapeutic, IPF, ALK5