Linezolid-mediated Prevention of Fibroblast Activation and Tissue Fibrosis via Mitochondrial Translation Inhibition.

OBJECTIVE: Beyond its role as a ribosome-targeting antibiotic, linezolid was recently shown to modulate immune responses by inhibiting mitochondrial translation. Because mitochondrial dysfunction is implicated in various fibrotic diseases, including systemic sclerosis (SSc), this study aimed to evaluate the antifibrotic potential of linezolid and delineate its underlying mechanisms in SSc. METHODS: The effects of linezolid on fibrotic tissue remodeling were assessed using multiple experimental systems: human dermal fibroblasts, human macrophages, three-dimensional SSc skin equivalents (SScSE), the murine model of sclerodermatous chronic graft-versus-host disease (sclGvHD) and precision-cut skin slices (PCSS) obtained from patients with SSc, RNA sequencing, immunofluorescence, Western blot, and histology. Mitochondrial function was evaluated using Seahorse assays alongside mitochondrial protein synthesis assessments. RESULTS: Linezolid inhibited TGFβ-induced fibroblast activation in cultured human fibroblasts, SScSE, sclGvHD mice, and SSc-PCSS, as demonstrated by reversal of profibrotic gene expression programs, downregulation of TGFβ, WNT, and JAK-STAT signaling, and reductions in αSMA expression or stress fiber formation, which led to reduced collagen deposition and ameliorated skin or lung fibrosis in vivo. Mechanistically, linezolid induced metabolic alterations by inhibiting mitochondrial translation in fibroblasts to hamper the oxidative phosphorylation, reduce the NAD(+)/NADH ratio, and down-regulate glycolysis, an essential metabolic pathway for fibroblast activation. CONCLUSION: This study provides the first evidence that inhibiting mitochondrial translation with linezolid ameliorates fibrotic tissue remodeling. Because linezolid is already clinically approved as a reserve antibiotic, these findings hold translational promise and support the use of linezolid as a novel treatment for fibrotic disorders after further validation.