Scalable Production of Multi-Source Hybrid Biomimetic Exosomes Co-Loaded with Therapeutic Plasmids for Treatment of High-Altitude Pulmonary Edema

Exosomes show therapeutic promise but face limitations in simultaneous targeting and potency. While hybrid exosome strategies combining multiple cell components can address this, current methods rely on native exosome extraction and drug loading, suffering from low yield and encapsulation efficiency. We developed a biological origin-based hybrid biomimetic exosome (BOB-HBE) platform that synthetically assembles exosomes using parent cell-derived components through a water/oil/water emulsion method, overcoming production bottlenecks. For high-altitude pulmonary edema (HAPE) treatment, we engineered hybrid exosomes combining: (1) vascular endothelial cell membranes for lung targeting, (2) mesenchymal stem cell factors for regeneration, and (3) eNOS-encoding plasmid DNA to restore nitric oxide signaling. The BOB-HBE platform achieved >150-fold higher production yield than natural exosome isolation while enhancing pulmonary endothelial specificity. In HAPE models, these hybrid exosomes demonstrated triple therapeutic effects: restoring NO bioavailability, inhibiting pathogenic HIF-1α/TGF/Smad1/5 signaling, and preventing endothelial-mesenchymal transition and vascular remodeling. Consequently, they significantly attenuated HAPE progression by addressing both molecular pathways and tissue-level pathology. This study establishes a scalable platform for constructing multifunctional exosome mimetics that maintain native exosome advantages while solving key production challenges. The cell-origin-informed design strategy offers a versatile approach for targeted therapy in pulmonary and vascular disorders, with potential clinical translation advantages.