Aspirin induces glycolytic reprogramming via lysine salicylation of ALDOA

Aspirin (ASA), a classical nonsteroidal anti-inflammatory drug, is known to inhibit glycolysis in cancer cells; however, the underlying mechanism remain incompletely understood. Here, we identify lysine salicylation (Ksa) as an ASA-dependent protein modification that suppresses glycolysis and proliferation in colorectal cancer cells. Specifically, ASA causes Ksa on the key glycolytic enzyme aldolase A (ALDOA), markedly reducing its enzymatic activity without altering protein level. Metabolomic analysis revealed that ASA impairs the conversion of fructose-1,6-bisphosphate (F16BP) to glyceraldehyde-3-phosphate (GAP), thereby disrupting glycolytic flux. In a xenograft mouse model, ASA administration not only inhibited tumor growth but also enhanced ALDOA Ksa level and blocked F16BP-to-GAP conversion. Besides, site-directed mutagenesis and genetic code expansion experiments confirmed that ALDOA K13sa is critical for ASA-mediated inhibition of glycolysis and cancer cell proliferation. Together, this study uncovers Ksa as an innovative mechanism by which ASA affects glycolysis and cancer cell proliferation, providing further insights into its potential role in cancer prevention and therapy.