Hepatocellular carcinoma (HCC) remains a highly prevalent malignancy worldwide with limited therapeutic options and frequent resistance to treatment, necessitating the development of targeted pharmacological interventions.

Although artesunate, a derivative of the widely used antimalarial drug artemisinin, has emerged as a promising anticancer candidate, the molecular basis underlying its antitumor activity in HCC has remained unclear.

Building on their previous study that identified glucosylceramidase (GBA) as a direct target of artesunate, this study published in Genes & Diseases by researchers from China Academy of Chinese Medical Sciences and Fujian University of Traditional Chinese Medicine elucidates the structural basis of this interaction and the downstream mechanisms responsible for its anticancer effects.

Using a combination of computational modelling, biochemical analyses, cellular assays, and in vivo experiments, the researchers demonstrated that artesunate markedly suppressed HCC cell proliferation while promoting apoptosis, with HepG2 cells exhibiting greater sensitivity than MHCC-97H cells.

The antitumor effects were associated with disruption of sphingolipid homeostasis, accumulation of glucosylceramide-related metabolites, mitochondrial dysfunction, and activation of caspase-dependent apoptotic signalling pathways.

Mechanistically, network and experimental analyses identified a GBA–ceramide–CTSD–α-syn–BID–BAX signalling axis that mediates artesunate-induced apoptosis in HCC.

Artesunate inhibits GBA, disrupting ceramide metabolism, impairing CTSD maturation, and promoting α-syn accumulation, which activates mitochondrial apoptosis through BID cleavage, BAX upregulation, and caspase activation.

Rescue experiments with ceramide supplementation or α-syn inhibition confirmed the central role of this pathway in artesunate's anticancer activity.

Through integrated computational and experimental analyses, the authors identified Y313, E340, and N396 as critical binding residues within the GBA active site.

Site-directed mutagenesis, particularly the E340R and N396R mutations, significantly impaired artesunate binding and reduced GBA enzymatic activity, thereby abolishing artesunate's ability to regulate downstream apoptotic signalling and establishing a direct mechanistic link between target engagement and therapeutic activity.

Collectively, this study provides the first detailed structural and mechanistic characterisation of how artesunate directly targets GBA in HCC, identifying the GBA–ceramide–CTSD–α-syn–BID–BAX signalling axis as a key mediator of mitochondrial apoptosis.

These findings establish GBA as a promising therapeutic target and advance understanding of artesunate's anticancer activity, supporting the rational development of GBA-targeted therapeutic strategies for HCC.

While the proposed mechanism was validated in HCC cell lines and supported by in vivo studies, further validation in patient-derived organoids and xenograft models, together with comprehensive long-term safety assessments, will be essential to confirm its clinical relevance and support translation toward HCC therapy.

Source: Compuscript Ltd