NeuroD1 gene therapy inhibits glioma growth and extends life span through in vivo reprogramming approach.

Glioblastoma(GBM), a highly aggressive primary brain tumor characterized by rapid progression, frequent recurrence, and limited clinical options, remains one of the most lethal central nervous system malignancies. Here, we report a gene therapy strategy to treat glioma utilizing NeuroD1, a neurogenic transcription factor with demonstrated capacity to reprogram both glial cells and GBM cells into neuronal lineages. We developed a self-complementary adeno-associated virus (scAAV) vector, scAAV6-NeuroD1, and evaluated its therapeutic potential across in vitro and in vivo GBM models, including multiple GBM cell lines, patient-derived organoids, and orthotopic models in immunodeficient mice. Our findings reveal that scAAV6-NeuroD1 preferentially infects glioma cells and induces dual therapeutic effects by simultaneously inhibiting glioma cell proliferation and inducing neuronal reprogramming. Importantly, scAAV6-NeuroD1-treated mice with orthotopic GBM transplants exhibited reduced tumor burden, infiltration of T cells into the glioma, attenuated tumor-induced body weight loss, and dose-dependent survival extension. Analysis of published patient datasets further revealed that high NeuroD1 expression level correlates with improved overall survival and lower tumor malignancy grade. Together, these findings position scAAV6-NeuroD1 as a promising therapeutic candidate, potentially redefining the therapeutic landscape for GBM.