嵌合抗原受体T细胞衍生的细胞外囊泡:一种潜在的癌症治疗方法。

Extracellular Vesicles Derived from Chimeric Antigen Receptor-T Cells: A Potential Therapy for Cancer.

作者信息

Aharon Anat, Horn Galit, Bar-Lev Tali Hana, Zagagi Yohay Einav, Waks Tova, Levin Maya, Deshet Unger Naamit, Avivi Irit, Globerson Levin Anat

机构信息

Hematology Research Laboratory for Extracellular Vesicles, Hematology Division, Tel-Aviv Sourasky Medical Center, Tel Aviv, Israel.

Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

出版信息

Hum Gene Ther. 2021 Oct;32(19-20):1224-1241. doi: 10.1089/hum.2021.192.

Abstract

Chimeric antigen receptor (CAR)-T cells are genetically engineered T cells, directed against a tumor-associated antigen. Extracellular vesicles (EVs) derived from CAR-T cells (CAR-T EVs) may preserve CAR-T activity and overcome one of the major obstacles responsible for CAR-T cell failure in patients with solid tumors. This study aimed to compare CAR-T EVs with their parental cells and explore their cell penetration and cytotoxic activity. Anti-HER-2 CARs were stimulated with specific target cells. EVs were isolated from the cell media and characterized for their content and functions. We found that CAR-T EVs contained a mixture of small and large EVs. Stimulated anti-HER-2 CAR-T EVs expressed lower cytokine levels compared with their parental CAR-T cells (such as interferon gamma). Higher levels of granzyme B were found in CAR-T EVs (≥20 × ) compared with EVs from unstimulated cells ( < 0.001). Anti-HER-2 CAR-T EVs bound and penetrated specifically into HER-2 expressing target cells. Similar cytotoxic effects measured by caspase-3/7 activity were found in CAR-T cells and their derived EVs. However, while the CAR-T cells induced massive apoptosis during the first 24 h, CAR-T EVs required 60 - 90 h. In summary, CAR-T EVs provide a novel potent immunotherapy approach that may be effective against solid tumors.

摘要

嵌合抗原受体(CAR)-T细胞是经过基因工程改造的T细胞,可靶向肿瘤相关抗原。源自CAR-T细胞的细胞外囊泡(CAR-T EVs)可能保留CAR-T活性,并克服实体瘤患者中导致CAR-T细胞失效的主要障碍之一。本研究旨在比较CAR-T EVs与其亲代细胞,并探索它们的细胞穿透能力和细胞毒性活性。用特异性靶细胞刺激抗HER-2 CARs。从细胞培养基中分离出细胞外囊泡,并对其内容物和功能进行表征。我们发现CAR-T EVs包含大小不同的细胞外囊泡混合物。与亲代CAR-T细胞相比,受刺激的抗HER-2 CAR-T EVs表达的细胞因子水平较低(如干扰素γ)。与未受刺激细胞的细胞外囊泡相比,CAR-T EVs中发现更高水平的颗粒酶B(≥20倍)(P<0.001)。抗HER-2 CAR-T EVs特异性结合并穿透表达HER-2的靶细胞。通过caspase-3/7活性测量发现,CAR-T细胞及其衍生的细胞外囊泡具有相似的细胞毒性作用。然而,虽然CAR-T细胞在最初24小时内诱导大量细胞凋亡,但CAR-T EVs需要60-90小时。总之,CAR-T EVs提供了一种新型有效的免疫治疗方法,可能对实体瘤有效。

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