Enhancing precision targeting of ovarian cancer tumor cells in vivo through extracellular vesicle engineering

Abstract Extracellular vesicles (EVs) function as natural mediators of intercellular communication, secreted by cells to facilitate cell–cell signaling. Due to their low toxicity, immunogenicity, biodegradability, and potential to encapsulate therapeutic drugs, EVs hold significant therapeutic promise. Nevertheless, their limited targeting ability often diminishes their therapeutic impact. Therefore, enhancing EVs by incorporating targeting units onto their membranes could bolster their targeting capabilities, enabling them to accumulate in specific cells and tissues. In this study, we engineered EVs to fuse ephrin‐B2 with the EV membrane protein LAMP2b. This modification aimed to direct the engineered EVs toward the ephrin‐B4 receptor expressed on the surface of ovarian cancer cells. The engineered EVs retained their inherent properties, including size, expression of EV membrane proteins, and morphology, upon isolation. In vitro experiments using real‐time imaging revealed that EVs engineered with the ephrin‐B2 ligand exhibited substantial internalization and uptake by ovarian cancer cells, in stark contrast to native EVs. In vivo, the engineered EVs carrying the ephrin‐B2 ligand effectively targeted ovarian cancer cells, surpassing the targeting efficiency of control EVs. This innovative approach establishes a novel targeting system, enhancing the uptake of EVs by ovarian cancer cells. Our findings underscore the potential of using EVs to target cancer cells, thereby enhancing the effectiveness of anti‐cancer therapies while minimizing off‐target effects and toxicity in normal cells and organs.