Abstract
<title>Abstract</title> <p>By virtue of inherent biocompatibility and delivery efficiency, extracellular vesicles (EVs) are regarded as a novel and promising drug delivery system (DDS) for disease treatment. In this study, we designed and generated engineered HEK293T cells (HEK919) featuring an inducible Tet-On® 3G system to co-express a uPAR-targeting domain (ATF-uPA), TNF-related apoptosis-inducing ligand(TRAIL), and the Herpes Simplex Virus Thymidine Kinase (HSV-TK) suicide gene. We confirmed that both the HEK919 cells and secreted EVs significantly inhibited proliferation and induced apoptosis in U87MG and U251 glioma cells in the presence of ganciclovir (GCV). Furthermore, the killing effect of HEK919 EVs was demonstrated to be dose- and time-dependent. In murine models of glioblastoma, the engineered EVs specifically accumulated in uPAR-positive tumor tissues and exhibited superior tumor growth suppression compared to HEK919 cell therapy. In summary, we establishes a targeted, inducible EV system that synergizes TRAIL- and suicide gene-mediated killing, offering a translatable and effective approach for treating uPAR-positive glioblastoma.</p>