Glutaminolysis is required for transforming growth factor-β1–induced myofibroblast differentiation and activation

Abstract

Myofibroblasts participate in physiological wound healing and pathological fibrosis. Myofibroblast differentiation is characterized by the expression of α-smooth muscle actin and extracellular matrix proteins and is dependent on metabolic reprogramming. In this study, we explored the role of glutaminolysis and metabolites of TCA in supporting myofibroblast differentiation. Glutaminolysis converts Gln into α-ketoglutarate (α-KG), a critical intermediate in the TCA cycle. Increases in the steady-state concentrations of TCA cycle metabolites including α-KG, succinate, fumarate, malate, and citrate were observed in TGF-β1-differentiated myofibroblasts. The concentration of glutamate was also increased in TGF-β1-differentiated myofibroblasts compared with controls, whereas glutamine levels were decreased, suggesting enhanced glutaminolysis. This was associated with TGF-β1-induced expression of the glutaminase (GLS) isoform, GLS1, which converts Gln into glutamate, at both the mRNA and protein levels. The stimulation of GLS1 expression by TGF-β1 was dependent on both SMAD3 and p38 mitogen-activated protein kinase activation. Depletion of extracellular Gln prevented TGF-β1-induced myofibroblast differentiation. The removal of extracellular Gln postmyofibroblast differentiation decreased the expression of the profibrotic markers fibronectin and hypoxia-inducible factor-1α and reversed TGF-β1-induced metabolic reprogramming. Silencing of GLS1 expression, in the presence of Gln, abrogated TGF-β1-induced expression of profibrotic markers. Treatment of GLS1-deficient myofibroblasts with exogenous glutamate or α-KG restored TGF-β1-induced expression of profibrotic markers in GLS1-deficient myofibroblasts. Together, these data demonstrate that glutaminolysis is a critical component of myofibroblast metabolic reprogramming that regulates myofibroblast differentiation.

Succinate receptor mediates intestinal inflammation and fibrosis

“…Interestingly, we found that the absence of this receptor reduced the pro-fibrotic effects of TGFβ in these cells. A recent study shows increased levels of succinate in fibroblasts treated with TGFβ, 30 but our results showing no increase in calcium levels after treatment of fibroblasts with TGFβ makes this possibility unlikely. We propose that Sucnr1 exacerbates the response of fibroblasts to pro-fibrotic stimuli as it promotes the response of immune cells to proinflammatory agents.…”

MiRNAs and mRNAs Analysis during Abdominal Preadipocyte Differentiation in Chickens

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Methods for assessing mitochondrial quality control mechanisms and cellular consequences in cell culture

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