Disruption of the NDC80–Nek2 axis suppresses glioblastoma stemness and enhances therapeutic efficacy

Glioblastoma (GBM), the most aggressive primary brain tumor, demonstrates resistance to standard therapies, primarily due to the presence of GBM stem cells (GSCs), the mechanisms of which remain poorly understood. To identify therapeutic targets for GSCs, we utilized multiple datasets to screen for differentially expressed pathways and potential targets. The prognostic and survival characteristics of the potential target NDC80 were analyzed using the TCGA and CGGA datasets. The molecular function of NDC80 was evaluated through western blot, RT-qPCR, limiting dilution assays, sphere-forming assays, cell proliferation, and cell cycle analysis. Orthotopic GBM xenograft models were established to assess the anti-GBM effects of NDC80 inhibition. Co-immunoprecipitation assays were performed to confirm the interaction between NDC80 and Nek2 after treatment with the NDC80 inhibitor TAI-1. We performed an integrated analysis of multiple datasets and identified significant differences in cell cycle–related pathways between GSCs and non-GSCs. Among these, NDC80, a core kinetochore protein crucial for accurate chromosome segregation during the G2/M phase of the cell cycle, was found to be markedly overexpressed in GSCs. Analysis of public databases revealed that high NDC80 expression is associated with poor patient prognosis and aggressive clinical features. Functional studies demonstrated that silencing NDC80 suppresses GSC stemness and tumorigenic potential, as evidenced by reduced self-renewal and proliferation in vitro, induction of G2/M phase arrest, and impaired tumor growth in an orthotopic brain tumor xenograft model. Furthermore, we showed that TAI-1, a small-molecule inhibitor that disrupts the NDC80–Nek2 interaction, effectively suppresses GSC proliferation and tumorigenesis, leading to a significant extension of survival in mice bearing orthotopic GBM tumors. Our findings highlight NDC80 as a key regulator of GBM pathogenesis and a promising prognostic biomarker and therapeutic target.