Ferroptosis-associated immune modulation enhances temozolomide efficacy in experimental glioblastoma: potential implications for T-cell infiltration and tumour microenvironment dynamics

Glioblastoma multiforme (GBM) remains a lethal and aggressive malignancy with limited response to standard therapies. This study hypothesized that artesunate, a ferroptosis inducer, enhances the cytotoxic and immunomodulatory efficacy of standard anticancer agents through mechanisms involving oxidative stress-mediated ferroptotic cell death and lipid peroxidation, influencing hypoxia-inducible factor-1α, insulin-like growth factor-1 (IGF-1), and leptin pathways. These pathways collectively regulate tumor proliferation, angiogenesis, and immune evasion; hence, their modulation may sensitize GBM to therapy. The study evaluated the therapeutic potential of artesunate in combination with anticancer drugs using in vitro GL261 glioblastoma cells and in vivo subcutaneous GL261 models. Clinically relevant agents representing distinct mechanisms, temozolomide, cyclophosphamide, paclitaxel, imatinib, and thymoquinone, were tested. Tumor volume, body weight, and serum biochemical parameters (IL-6, IGF-1, HIF-1α, and leptin) were analyzed, along with CD3 + T cell infiltration by immunohistochemistry. Combination treatments, particularly artesunate with temozolomide or paclitaxel, produced marked tumor regression compared with controls. Serum biomarker modulation and enhanced infiltration of CD3 + T cells indicated activation of ferroptotic and immune-mediated anti-tumor mechanisms, with minimal systemic toxicity. Artesunate potentiates the antitumor efficacy of standard chemotherapeutic agents through mechanisms involving ferroptosis and immune modulation, promoting a tumor-suppressive microenvironment. These findings support further mechanistic and translational studies toward developing ferroptosis-driven multimodal therapies for resistant glioblastomas.