Angiopep-2-decorated bacterial outer membrane vesicles penetrate the blood-brain barrier for glioblastoma chemo-immunotherapy.

Glioblastoma (GBM) therapy remains one of the most formidable challenges in oncology because of the blood-brain barrier (BBB), immunosuppressive tumor microenvironment, and drug resistance. To overcome these challenges, we designed engineered angiopep-2-decorated bacterial outer membrane vesicles for targeted drug delivery across the BBB. Using a novel autotransporter-based (AIDA-I) genetic engineering approach, we developed a novel platform via straightforward genetic modification of E. coli with our designed pAIDA1-ANG plasmid to stably display Angiopep-2 on OMVs (OMV-^ANG) for targeted BBB penetration via LDL receptor-related protein 1 (LRP1) receptor-mediated transcytosis. Our in vitro BBB model and real-time in vivo imaging confirmed the BBB penetration and transcytosis of OMV-^ANG and doxorubicin-loaded OMV-^ANG, with accelerated brain accumulation within 2-4 h post-injection and sustained retention for 6 h. In orthotopic GBM models, systemic DOX-OMV-^ANG administration extended survival, induced potent tumor suppression via DOX-induced apoptosis and OMV-^ANG-mediated immunomodulation, and triggered sustained IFN-γ elevation with macrophage and CD8 + T-cell recruitment. This engineered OMV-^ANG platform shows promise in overcoming critical therapeutic barriers in glioblastoma and warrants further investigation as a versatile delivery system for diverse CNS therapeutics.