Preclinical demonstration of the efficacy and mechanisms of antitumor activity of dihydroartemisinin in cholangiocarcinoma xenograft: Induction of autophagy and reprogramming of metabolism

Abstract

Cholangiocarcinoma (CCA) is a highly aggressive malignancy of the biliary tract with limited therapeutic options and poor prognosis. Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, has shown promising anticancer activity across various tumor types. In this study, we evaluated the anti-tumor efficacy and metabolic impact of DHA in a preclinical xenograft model of CCA. Human CCA cells were subcutaneously implanted into immunodeficient mice to establish tumor xenografts, which were then orally treated with various concentrations of DHA. Tumor volume and body weight were monitored throughout the treatment period. No signs of systemic toxicity or significant weight loss were noted in our treated mice. Our results demonstrated that DHA treatment at 50 mg/kg significantly reduced tumor volume compared to untreated controls. Histopathological analysis, immunohistochemistry, molecular assays, and metabolomic profiling were performed to assess treatment response and underlying mechanisms. Mechanistically, DHA inhibited CCA cell proliferation by inducing apoptosis and activating the autophagy pathway, as confirmed by increased apoptotic markers and autophagy-related proteins in tumor tissues. In addition, metabolomic analysis revealed that DHA significantly reduced the levels of key metabolites, including pyridoxine, D -threitol, and succinate, compared to control mice, indicating disruption of metabolic pathways critical for tumor growth and survival. These findings highlight the dual anti-cancer mechanisms of DHA, combining cell death induction and metabolic interference, and support further clinical investigation of DHA as a novel therapeutic agent for CCA.