Abstract
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.