Chimeric biohybrid nanovesicles induce immunogenic cell death for targeted and immune-potentiated glioblastoma therapy.

Glioblastoma (GBM) is shielded by both the blood-brain barrier (BBB) and an immunosuppressive tumor microenvironment. Here, we develop a chimeric biohybrid nanovesicle (BEV-RVG29-PTX) that integrates viral tropism, bacterial vesiculation, and chemotherapeutic cytotoxicity into a single genetically programmable platform. Genetic fusion of rabies virus glycoprotein 29 (RVG29) to the AIDA1 autotransporter translocator domain enables robust, autonomous surface expression on bacterial extracellular vesicles (BEVs) without the need for chemical conjugation. The BEV-RVG29-PTX drives receptor-dependent BBB transcytosis and achieves efficient glioma accumulation. Encapsulated paclitaxel (PTX), otherwise restricted by BBB impermeability, is effectively delivered to intracranial tumors and induces reactive oxygen species-driven immunogenic cell death. Bone marrow-derived dendritic cells immune-activation experiments further confirmed an approximately 2-fold increase in CD80/CD86 activation. Synergizing with the pathogen-mimetic characteristics of BEVs, these signals also elicit an approximately 2-fold increase in intratumoral CD8⁺ T-cell infiltration, overcome immune exclusion, and achieve durable tumor control with extended survival in orthotopic GBM models. Accordingly, this virus-bacteria-drug biohybrid strategy enables targeted brain delivery while simultaneously amplifying antitumor immunity, offering a promising and translatable approach for GBM treatment.