Inhibition of GOT1 potentiates ferroptosis and exacerbates cell death in acute myeloid leukemia

Drug resistance, poor treatment response, and low long-term survival rates are major obstacles to the successful long-term treatment of acute myeloid leukemia (AML), a heterogeneous hematologic malignancy. A deeper understanding of the molecular mechanisms underlying AML is therefore essential. Glutamic oxaloacetic transaminase 1 (GOT1), a key metabolic enzyme, has emerged as a potential regulator of AML progression, though its precise role remains unclear. This study aimed to investigate the expression profile, functional significance, and mechanistic involvement of GOT1 in AML. The expression levels of GOT1 in AML patient samples and cell lines were quantified by RT-qPCR and western blotting. Genetic knockdown (shRNA) and pharmacological inhibition (compound 2C) were employed to suppress GOT1 activity. Functional assays, including CCK-8 assay and Annexin V/PI staining, evaluated proliferation and apoptosis of AML cells. Multi-omics analyses (transcriptomics, proteomics, and metabolomics) were performed to delineate the mechanistic landscape of GOT1 inhibition. Reactive oxygen species (ROS) levels, glutathione (GSH) content, malondialdehyde (MDA) levels, and nicotinamide adenine dinucleotide phosphate (NADPH) levels in AML cells treated with compound 2C were measured. The combined effects of compound 2C and ferroptosis inducers (RSL3 or FIN56) were evaluated using the CCK-8 assay and Annexin V/PI staining. Synergy scores were calculated by SynergyFinder analysis based on the HSA reference model. The expression of GOT1 was significantly upregulated in AML patients and cell lines compared to controls. Inhibition of GOT1 suppressed AML proliferation and induced cell death via triggering ferroptosis. Multi-omics profiling revealed perturbations in redox homeostasis and iron metabolism pathways upon GOT1 suppression. Notably, GOT1 inhibition (compound 2C) combined with ferroptosis inducers (RSL3 or FIN56) synergistically reduced AML cell viability and induced apoptosis. Our findings demonstrate that GOT1 drives AML progression by modulating ferroptosis susceptibility. Targeting GOT1, either alone or in combination with ferroptosis inducers, represents a novel therapeutic strategy to overcome resistance and improve outcomes in AML.