Abstract 6583: Optimized production, biodistribution, and targeted delivery of T cell-derived extracellular vesicles for liver cancer therapy

Abstract Background: Extracellular vesicles (EVs) have emerged as potent mediators of physiological and pathological processes. By transporting bioactive molecules such as proteins and RNAs, EVs play a pivotal role in maintaining tissue and organ homeostasis. This unique functionality positions EVs as highly promising therapeutic agents. Notably, their ability to penetrate solid tumors and deliver targeted treatments underscores their potential in oncology. However, the preclinical use of EVs requires the development of robust and scalable production methods. This study aimed to establish standardized protocols for efficient EV production while evaluating their tissue and organ targeting capabilities, with a focus on achieving precise and efficient delivery to the liver. Furthermore, the potential application of EVs in minimally invasive interventions, such as portal vein embolization, opens new avenues for therapeutic delivery to liver tissues. Methods: EVs were isolated from T cells derived from Buffalo rat spleens and activated with concanavalin A. After 48 hours of culture, EVs were precipitated from the cell culture supernatant. The size and concentration of the isolated EVs were assessed using NanoSight technology. EVs were labeled with IVISense 750 dye and resuspended in either a saline solution or an oil-based contrast agent. These labeled EVs were administered to tumor-bearing rats via three routes: tail vein, portal vein, and intratumoral injections. Biodistribution of the EVs in liver tissue and other organs was analyzed using in vivo and ex vivo imaging. Results: Portal vein injection of saline-suspended EVs demonstrated superior liver targeting, achieving rapid and efficient accumulation in the liver. Intratumoral injection successfully delivered EVs to localized tumor regions, while tail vein injection, despite a slight delay, also enhanced liver biodistribution. Imaging results revealed a strong tropism of EVs for tumor tissues. Interestingly, systemic administration via the tail vein resulted in reduced kidney signals, suggesting urinary clearance of EVs within 72 hours. Conclusions: This study established a reproducible, standardized protocol for activating spleen-derived T cells, enabling large-scale production of EVs suitable for in vivo applications. The findings highlight the potential of EVs as effective delivery vehicles for therapeutic cargo, particularly for liver-targeted and tumor-specific treatments. Moreover, EV application in minimally invasive interventions, such as portal vein embolization, could further enhance targeted delivery to liver tissues. By optimizing both production and delivery methods, this research lays a foundation for advancing EV-based strategies in cancer therapy, paving the way for future clinical applications. Citation Format: Aminah Jaffery, Andrea C. Cortes, Kiyoyuki Minamiguchi, Simone Anfossi, Rony Avritscher. Optimized production, biodistribution, and targeted delivery of T cell-derived extracellular vesicles for liver cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6583.