AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment.

High-grade gliomas are among the most treatment-refractory solid tumors, with profound immunosuppression, diffuse infiltration, and resistance to existing immunotherapies. Interferons (IFNs) have potent anti-tumor and immunostimulatory activities, but systemic IFN therapies have failed in solid tumors due to poor pharmacokinetics and dose-limiting toxicities. Here, we introduce vectorized IFNs (recombinant adeno-associated virus [AAV] gene therapies enabling sustained, localized IFN expression within tumors) as a new class of biologics for high-grade glioma treatment. We engineered AAV vectors expressing human IFNα1, IFNβ, IFNγ, or combinations thereof and demonstrated potent, selective anti-tumor activity in patient-derived glioblastoma organoids while sparing healthy brain cells. Transcriptomic profiling revealed a durable, IFN-specific response in treated tumor cells. AAV-hIFNβ was advanced to orthotopic patient-derived and cell line-derived xenograft models, where intratumoral convection-enhanced delivery produced complete tumor clearance and significantly prolonged survival. Spatial transcriptomic and histologic analyses showed tumor-localized IFN expression, immune activation, and apoptosis-mediated tumor regression. To enable clinical translation, we developed SRN-101, a clinical-grade AAV-hIFNβ construct with optimized potency and manufacturability, which achieved superior expression and in vivo efficacy vs. the research-grade vector. These findings establish AAV-vectorized cytokines as a durable, locally delivered immuno-gene therapy platform for high-grade gliomas.