4-Octyl itaconate inhibits aerobic glycolysis by targeting GAPDH to exert anti-inflammatory effects

• Published in: Nature communications Vol. 10; no. 1; pp. 5091 - 11
• Main Authors: Liao, Shan-Ting, Han, Chao, Xu, Ding-Qiao, Fu, Xiao-Wei, Wang, Jun-Song, Kong, Ling-Yi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 08.11.2019; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/1; 13/21; 38; 38/77; 42; 42/109; 631/250/2504/342; 631/250/256/2516; 631/443/319; 631/45/320; 64; 64/60; 82/58; Alkylates; Alkylation; Animals; Antimetabolites - pharmacology; Cysteine - drug effects; Cysteine - genetics; Cysteine - metabolism; Deoxyglucose - pharmacology; Down-Regulation; Endotoxemia - immunology; Enzymatic activity; Enzyme activity; Enzymes; Glucose - metabolism; Glyceraldehyde-3-phosphate dehydrogenase; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - antagonists & inhibitors; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - drug effects; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - metabolism; Glycolysis; Glycolysis - drug effects; Humanities and Social Sciences; Inflammation - immunology; Inflammatory diseases; Interleukin-1beta - drug effects; Interleukin-1beta - immunology; Lethality; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Macrophage Activation - immunology; Macrophages; Macrophages - drug effects; Macrophages - immunology; Macrophages - metabolism; Metabolites; Mice; multidisciplinary; Nitric Oxide Synthase Type II - drug effects; Nitric Oxide Synthase Type II - immunology; Oxidative phosphorylation; Oxidative Phosphorylation - drug effects; Phosphorylation; RAW 264.7 Cells; Science; Science (multidisciplinary); Sesquiterpenes - pharmacology; Succinates - pharmacology; Therapeutic applications
• DOI: 10.1038/s41467-019-13078-5
• ISSN: 2041-1723

Activated macrophages switch from oxidative phosphorylation to aerobic glycolysis, similar to the Warburg effect, presenting a potential therapeutic target in inflammatory disease. The endogenous metabolite itaconate has been reported to regulate macrophage function, but its precise mechanism is not clear. Here, we show that 4-octyl itaconate (4-OI, a cell-permeable itaconate derivative) directly alkylates cysteine residue 22 on the glycolytic enzyme GAPDH and decreases its enzyme activity. Glycolytic flux analysis by U 13 C glucose tracing provides evidence that 4-OI blocks glycolytic flux at GAPDH. 4-OI thereby downregulates aerobic glycolysis in activated macrophages, which is required for its anti-inflammatory effects. The anti-inflammatory effects of 4-OI are replicated by heptelidic acid, 2-DG and reversed by increasing wild-type (but not C22A mutant) GAPDH expression. 4-OI protects against lipopolysaccharide-induced lethality in vivo and inhibits cytokine release. These findings show that 4-OI has anti-inflammatory effects by targeting GAPDH to decrease aerobic glycolysis in macrophages. Redirection of the TCA cycle intermediate aconitate to itaconate production has anti-inflammatory effects. Here the authors show that the itaconate derivative 4-octyl-itaconate is anti-inflammatory partly as a result of inhibiting GAPDH enzymatic activity and thereby glycolysis in macrophages.