Targeted Delivery of IL-12 Via CD19-Modified Extracellular Vesicles Enhances CAR-T Cell Efficacy Against Lymphoma

作者：Meijuan Huang, Lei Zhao, Dengju Li, Liang Huang, Jue Wang, Yuanyuan Zhang · 发表于：Blood · 年份：2023 · DOI：10.1182/blood-2023-180860 · 被引用次数：6 · 研究领域：CAR-T cell therapy research、Extracellular vesicles in disease

Introduction Chimeric Antigen Receptor T-Cell (CAR-T cell) Therapy faces challenges related to low potency and poor in vivo persistence. Incorporating cytokine signals, such as interleukin-12 (IL-12), has emerged as a promising strategy to enhance CAR-T cell functionality due to its potent immunostimulatory effects. However, clinical application of IL-12 is hindered by pleiotropic toxicity and limited bioavailability. Therefore, alternative agents or delivery systems are desired to exploit the benefits of IL-12 safely. This study investigated the therapeutic potential of a novel IL-12 delivery system utilizing extracellular vesicles (EVs) modified with the target antigen CD19 to specifically deliver IL-12 to CD19 CAR-T cells, thus augmenting their anti-tumor effects. Method IL-12-anchored extracellular vesicles (IL-12 EVs) were generated from engineered 293T cells expressing IL-12 on the cell membrane via GPI structure. Additionally, EVs co-expressing IL-12 and CD19 (CD19/IL-12 EVs) were produced from 293T cells co-expressing CD19 and IL-12. The physicochemical characteristics of the EVs were analyzed using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blot (WB). Binding and uptake efficiency of the EVs with CD19 CAR-T cells were validated using flow cytometry and imaging flow cytometry. In vitro experiments were conducted to investigate the effects of the EVs on the cytotoxic function, degranulation, cytokine secretion, and prolife...