Anlotinib Inhibits PFKFB3-Driven Glycolysis in Myofibroblasts to Reverse Pulmonary Fibrosis

Abstract

Idiopathic pulmonary fibrosis (IPF) is a fatal disease in which the normal alveolar network is gradually replaced by fibrotic scars. Current evidence suggests that metabolic alterations correlate with myofibroblast activation in IPF. Anlotinib has been proposed to have antifibrotic effects, but the efficacy and mechanisms of anlotinib against lung fibrosis have not been systematically evaluated. The antifibrotic effects of anlotinib were evaluated in bleomycin-induced mouse models and transforming growth factor-beta 1 (TGF-β1)-stimulated lung fibroblasts. We measured lactate levels, 2-NBDG glucose uptake and the extracellular acidification rate (ECAR) to assess glycolysis in fibroblasts. RNA-protein coimmunoprecipitation (RIP) and polysome analyses were performed to investigate novel mechanisms of glycolytic reprogramming in pulmonary fibrosis. We found that anlotinib diminished myofibroblast activation and inhibited the augmentation of glycolysis. Moreover, we show that PCBP3 posttranscriptionally increases PFKFB3 expression by promoting its translation during myofibroblast activation, thus promoting glycolysis in myofibroblasts. Regarding mechanism, anlotinib exerts potent antifibrotic effects by downregulating PCBP3, reducing PFKFB3 translation and inhibiting glycolysis in myofibroblasts. Furthermore, we observed that anlotinib had preventative and therapeutic antifibrotic effects on bleomycin-induced pulmonary fibrosis. Therefore, we identify PCBP3 as a protein involved in the regulation of glycolysis reprogramming and lung fibrogenesis and propose it as a therapeutic target for pulmonary fibrosis. Our data suggest that anlotinib has antifibrotic effects on the lungs, and we provide a novel mechanism for this effect. Anlotinib may constitute a novel and potent candidate for the treatment of pulmonary fibrosis.

Persistent hypoxia promotes myofibroblast differentiation via GPR‐81 and differential regulation of LDH isoenzymes in normal and idiopathic pulmonary fibrosis fibroblasts

“…Prior reports indicate that hypoxia can induce fibroblast differentiation in normal fibroblasts (Chen, Zhang, et al, 2021; Kottmann et al, 2015). These findings suggest that α‐SMA expression is modulated when control fibroblasts are exposed to hypoxic conditions.…”

Anlotinib suppresses lung adenocarcinoma growth via inhibiting FASN-mediated lipid metabolism

“…Here, however, our study provided the evidence that anlotinib could attenuate tumor growth through direct disturbance of lipid metabolism in LUAD cells. It has been shown that anlotinib could inhibit glycolysis in myofibroblasts to reverse pulmonary fibrosis, which indicates that anlotinib might be capable of interfering with nutrition metabolism in cancer cells, thereby eliciting killing effects (19). A recent study also showed that anlotinib could interfere with amino acid metabolism in colon cancer cells leading to impaired protein synthesis and disruption of energy supply (20), which indicates that the killing effect of anlotinib in cancer cells is linked to metabolism.…”

Metabolic reprogramming of pulmonary fibrosis

“…PFKFB3 expression can be increased by the secretion of TNF-α in lung fibroblasts, while PFKFB3 suppression effectively blocks TNF-α-induced aerobic glycolysis and lactate production in fibroblasts ( Xu et al, 2022 ). The anti-fibrotic effect of Anlotinib is demonstrated to inhibit PFKFB3-dependent glycolysis, which can be post-transcriptionally regulated by the RNA-binding protein PCBP3 ( Chen et al, 2021 ).…”