Expanding the toolbox of exosome-based modulators of cell functions

Exosomes are cell-derived extracellular vesicles and play important roles in mediating intercellular communi-cations. Due to their unique advantages in transporting a variety of biomolecules, exosomes have been emerging as a new class of nanocarriers with great potential for therapeutic applications. Despite advancements in loading chemotherapeutics and interfering RNAs into exosomes, active incorporation of protein molecules into exosomes remains challenging owing to their distinctive physicochemical properties and/or a lack of knowledge of cargo sorting during exosome biogenesis. Here we report the generation of a novel type of engineered exosomes with actively incorporated membrane proteins or soluble protein cargos, named genetically infused functionally tailored exosomes (GIFTed-Exos). Through genetic fusion with exosome-associated tetraspanin CD9, trans-membrane protein CD70 and glucocorticoid-induced tumor necrosis factor receptor family-related ligand (GITRL) could be displayed on exosome surface, resulting in GIFTed-Exos with excellent T-cell co-stimulatory activities. By genetically linking to a CD9-photocleavable protein fusion, fluorescent protein mCherry, apoptosis-inducing protein apoptin, and antioxidant enzyme catalase could be effectively packed into exosomes for light-controlled release. The generated GIFTed-Exos display notable in vitro and in vivo activities for delivering distinct types of protein cargos to target cells. As a possibly general approach, GIFTed-Exos provide new op-portunities to create exosomes with new functions and properties for biomedical research.

Automated summary

Exosomes, as cell-derived extracellular vesicles, are crucial for intercellular communication and have emerged as promising nanocarriers for therapeutic applications. By genetically engineering exosomes, a new type of exosomes called GIFTed-Exos with specific functions can be created to efficiently deliver different types of protein cargos to target cells.