癌症免疫治疗中治疗性细胞外囊泡（EVs）的发展取得了重大进展，使其成为细胞治疗的一种替代方法。在这项概念验证工作中，我们通过对EV产生的树突状细胞（DCs）进行基因工程，并引入aCD19 scFv和PD1，开发了双特异EV（BsEVs），用于同时靶向肿瘤抗原和阻断免疫检查点蛋白。我们发现这些双特异EVs（EVs-PD1-aCD19）在静脉注射后具有在高表达huCD19的实体肿瘤中积累的显著能力。此外，EVs-PD1-aCD19通过阻断PD-L1显著逆转了实体肿瘤的免疫局势。此外，EVs-PD1-aCD19还可以靶向在循环中存在的肿瘤来源EVs，从而防止在其他组织中形成原发性转移巢。我们的技术是一种基于双特异EV的癌症免疫治疗演示，可以启发针对具有不同表面抗原的各种类型的肿瘤的治疗，甚至是个体化的治疗。版权所有 © 2023作者。Elsevier Inc.发表并保留所有权利。

Advances in the development of therapeutic extracellular vesicles (EVs) for cancer immunotherapy have allowed them to emerge as an alternative to cell therapy. In this proof-of-concept work, we develop bispecific EVs (BsEVs) by genetically engineering EV-producing dendritic cells (DCs) with aCD19 scFv and PD1 for targeting tumor antigens and blocking immune checkpoint proteins simultaneously. We find that these bispecific EVs (EVs-PD1-aCD19) have an impressive ability to accumulate in huCD19-expressing solid tumors following intravenous injection. In addition, EVs-PD1-aCD19 can remarkably reverse the immune landscape of the solid tumor by blocking PD-L1. Furthermore, EVs-PD1-aCD19 can also target tumor-derived EVs in circulation, which prevents the formation of a premetastatic niche in other tissues. Our technology is a demonstration of bispecific EV-based cancer immunotherapy, which may inspire treatments against various types of tumors with different surface antigens and even a patient-tailored therapy.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.