GCLM lactylation mediated by ACAT2 promotes ferroptosis resistance in KRASG12D-mutant cancer

This data consists of raw immunoblotting images from the article titled "KGCLM lactylation mediated by ACAT2 promotes ferroptosis resistance in KRASG12D-mutant cancer". KRAS mutations drive tumorigenesis, but their role in ferroptosis regulation remains unclear. Here, we construct KRASWT and KRASG12D-mutant cancer cells and demonstrate that G12D-mutant cells exhibit increased viability and reduced ferroptosis upon RSL3 or erastin treatment. These cells show diminished lipid peroxidation and mitochondrial damage, indicating ferroptosis resistance. KRASG12D activates MEK-ERK signaling to phosphorylate LDHA, enhancing glycolysis and lactate production. Exogenous lactate supplementation similarly protects wild-type cells from ferroptosis. Mechanistically, G12D mutation-derived lactate induces Glutamate-cysteine ligase modifier (GCLM) lactylation, a process catalysed by Acetyl-CoA Acetyltransferase 2 (ACAT2). Inhibition of GCLM lactylation either through the mutation of the lactylation site or by knockdown of ACAT2 diminished the enzymatic activity of Glutamate-cysteine ligase (GCL) and suppressed Glutathione synthesis. Importantly, ACAT2 depletion overcomes ferroptosis resistance in KRASG12D-mutant tumors in vivo. Our findings reveal a KRASG12D-driven metabolic adaptation linking GCLM lactylation to ferroptosis resistance, proposing ACAT2 inhibition as a therapeutic strategy for KRAS-mutant cancers.