Antibody blockade of ENO1 attenuates TGF-β-linked metabolic reprogramming and suppresses myofibroblast differentiation in pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive disease driven by transforming growth factor-β (TGF-β)-mediated myofibroblast differentiation and metabolic reprogramming. While our previous work identified enolase-1 (ENO1) as a pro-fibrotic mediator, the precise mechanism by which it orchestrates fibrotic progression remains unclear. Here, we investigated whether the clinical-stage anti-ENO1 antibody HuL001 could mitigate the progression of pulmonary fibrosis by modulating TGF-β-associated metabolic reprogramming. The anti-fibrosis efficacy of HuL001 was evaluated in a murine bleomycin model, with treatment initiated during the fibrotic phase (Day 7). Fibrotic remodeling was assessed using histopathology analysis and measurements of collagen deposition and myofibroblast accumulation. In primary lung fibroblasts, we examined the effects of HuL001 on TGF-β-induced differentiation and contractility. Mechanistic studies focused on cell-surface translocation of ENO1, hypoxia-inducible factor-1α (HIF-1α) expression, glycolytic flux, and latent TGF-β activation. Administration of HuL001 significantly attenuated bleomycin-induced lung fibrosis and reduced collagen and myofibroblast accumulation. Ex vivo, HuL001 suppressed TGF-β-induced myofibroblast differentiation. We found that TGF-β promoted the translocation of ENO1 to the surface of lung fibroblasts, upregulated HIF-1α, and enhanced glycolysis and lactate production. Crucially, this metabolic shift accelerated the activation of latent TGF-β, creating a profibrotic amplification loop. By specifically targeting extracellular ENO1, HuL001 disrupted this cycle, resulting in concurrent reduction in HIF-1α expression, glycolytic flux, and TGF-β activation. Our findings demonstrate that HuL001 suppresses myofibroblast differentiation and attenuates the progression of pulmonary fibrosis by interrupting an ENO1-dependent metabolic feed-forward loop. These results validate extracellular ENO1 as a promising therapeutic target for IPF and provide a mechanistic rationale for the clinical development of HuL001.