PKLR-modulating compounds show promising effects for the treatment of hepatocellular carcinoma

The pyruvate kinase L/R (PKLR) enzyme was associated with the progression of hepatocellular carcinoma (HCC) and metabolic dysfunction-associated fatty liver disease (MAFLD) and identified as a potential drug target. Here, we first demonstrated the anti-HCC effects of recently developed six PKLR-modulating compounds, including SET135, SET156, SET158, SET159, SET171, and SET172 and JNK-IN-5A, a repurposed compound for modulation of PKLR based on systems biology. Compared with the multi-target kinase inhibitors sorafenib and regorafenib, PKLR-modulating compounds exhibited enhanced induction of cell cycle arrest mediated by p53 and increased levels of autophagy, apoptosis, and reduced invasiveness. Second, we performed global gene expression profiling using RNA sequencing and revealed the mechanism of action of each compound. Among these PKLR-modulating compounds, two exhibited unique pathways leading to necrotic cell death. The SET135 triggered autophagic necrotic cell death via p62/SQSTM1 induction, whereas the SET171 facilitated necrotic cell death by increasing cellular reactive oxygen species (ROS) and lipid peroxidation. Third, we validated that these PKLR-modulating compounds exert distinct cellular effects, showcasing stronger cell cycle arrest and apoptosis when compared to sorafenib and regorafenib based on systems biology analysis. Finally, we treated a HCC-transplanted mouse model with SET135 and SET171 for 21 days and showed that both compounds have superior anti-HCC therapeutic effects than sorafenib via apoptotic cell death. Our findings suggest that PKLR-modulating compounds, capable of inducing both apoptotic and necrotic cell death, represent promising candidates for novel HCC therapeutic agents.