Redirecting Monocyte Differentiation With Engineered Extracellular Vesicles for Glioma Immunotherapy

During glioma progression, monocytes abundantly infiltrate but primarily differentiate into immunosuppressive macrophages to promote tumor growth. Redirecting monocyte differentiation offers a compelling yet underexplored therapeutic opportunity. In this work, we found M1-polarized macrophage-derived extracellular vesicles (M1-EVs) efficiently induced monocytes to differentiate into anti-tumor macrophages via tumor necrosis factor alpha (TNF-α)-mediated signaling. Despite promising, the therapeutic efficacy of M1-EVs was constrained by insufficient glioma accumulation and CD47-mediated phagocytic inhibition. To address this challenge, we further engineered M1-EVs with dual-targeting specificity by genetically incorporating a tumor-directed chimeric antigen receptor (CAR) against IL13Rα2 or EGFRvIII together with CD47-blocking SIRPα variants. The resulting dual-targeting EVs (M1-CS-EVs) exhibited enhanced blood-brain barrier (BBB) penetration and glioma accumulation while locally disrupting CD47-SIRPα interactions. In three orthotopic glioma models, M1-CS-EVs elicited a potent anti-tumor immune response and enhanced tumor phagocytosis, significantly suppressing tumor growth while prolonging animal survival. Our findings establish a platform technology for directing monocyte differentiation toward anti-tumor phenotypes, offering a broadly applicable strategy for glioma treatment.