Endosymbiosis-inspired engineering of mitochondrial-enriched extracellular vesicles supercharges host mitochondria via the autophagy-lysosome pathway

Abstract Although the delivery of healthy mitochondria represents a promising strategy for the management of nonhealing wounds such as diabetic wounds, the optimal methods for efficient mitochondrial transfer and how the transferred mitochondria exert therapeutic effects remain largely unexplored. In this study, we identified mitochondrial and immunometabolic dysfunction in macrophages as pivotal drivers of delayed diabetic wound healing. Inspired by endosymbiosis, we engineered extracellular vesicles enriched in functional mitochondria (EVsMito) using a coextrusion strategy. EVsMito facilitated targeted delivery of functional mitochondria to diabetic macrophages and enhanced wound healing by reversing mitochondrial and immunometabolic dysfunction. Notably, transferred mitochondria colocalized with autophagosomes and were degraded via the autophagy–lysosome pathway, thereby improving host mitochondrial and immunometabolic functions. Ablation of autophagy‒lysosome fusion negated the therapeutic effects of EVsMito. This study provides an effective strategy for mitochondrial transfer and elucidates the fate of transferred mitochondria, providing new insights into mitochondrial transfer-based therapeutics for diabetic wound healing and other mitochondrial dysfunction-related diseases.