Li Guangchao, Zhang Qing, Han Zeping, Zhu Yangmin, Shen Huijuan, Liu Zhi, Zhou Zhao, Ding Wen, Han Siqi, He Jinhua, Yin Zhao, Zhou Jie, Ou Ruiming, Luo Min, Liu Shuang
Department of Hematology, Guangdong Second Provincial General Hospital, Guangzhou, China.
Department of Research and Development, Guangzhou Bio-gene Technology Co., Ltd, Guangzhou, China.
Front Oncol. 2021 Oct 27;11:734593. doi: 10.3389/fonc.2021.734593. eCollection 2021.
Chimeric antigen receptor T (CAR-T) cells are not effective in solid tumor treatment due to reduced invasion and expansion, and short survival time. This study aimed to explore whether interleukin (IL)-7 and CCR2b expression could improve GD2-CAR-T cell survival and infiltration in neuroblastoma and melanoma treatment. IL-7 and CCR2b were inserted into the classical second-generation CAR structure to construct 7×2b CAR. The 7×2b CAR-T cell phenotypes were evaluated by flow cytometry and the chemokine levels by ELISA. The 7×2b CAR-T cell migration and anti-tumor abilities were detected by Transwell assay and animal experiments . We report that compared with that of CAR-T cells, 7×2b CAR-T cell IL-7 secretion and CCR2b expression did not affect the T cell surface expression of CAR or CAR-T specificity and efficacy against tumor cells. The 7×2b CAR-T cells could induce IFN-γ secretion in GD2-positive tumor cells, killing them as well as conventional CAR-T cells. Moreover, IL-7 and CCR2b co-expression enhanced the 7×2b CAR-T cell survival and migration. Similar to conventional CAR-T, 7×2b CAR-T cells could also inhibit tumor growth and increase IFN-γ, Gzms-B, and IL-2 expression. Finally, unlike in mice injected with CAR-T cells, CD3 expression was the most abundant in the spleen and tumor tissues in mice injected with 7×2b CAR-T cells. Our study demonstrates that IL-7 and CCR2b co-expression in GD2-CAR-T cells exhibit stronger anti-tumor activity than classical second-generation CAR-T cells, shedding light on the potential novel GD2-positive neuroblastoma and melanoma treatment approach.
嵌合抗原受体T（CAR-T）细胞在实体瘤治疗中效果不佳，原因是其浸润和扩增能力降低以及存活时间较短。本研究旨在探讨白细胞介素（IL）-7和CCR2b的表达是否能改善GD2-CAR-T细胞在神经母细胞瘤和黑色素瘤治疗中的存活及浸润情况。将IL-7和CCR2b插入经典的第二代CAR结构中构建7×2b CAR。通过流式细胞术评估7×2b CAR-T细胞表型，用ELISA检测趋化因子水平。通过Transwell实验和动物实验检测7×2b CAR-T细胞的迁移和抗肿瘤能力。我们报告，与CAR-T细胞相比，7×2b CAR-T细胞的IL-7分泌和CCR2b表达不影响CAR在T细胞表面的表达，也不影响CAR-T细胞对肿瘤细胞的特异性和疗效。7×2b CAR-T细胞可诱导GD2阳性肿瘤细胞分泌IFN-γ，从而杀死这些细胞，其效果与传统CAR-T细胞相同。此外，IL-7和CCR2b共表达增强了7×2b CAR-T细胞的存活和迁移能力。与传统CAR-T细胞类似，7×2b CAR-T细胞也能抑制肿瘤生长，并增加IFN-γ、颗粒酶B和IL-2的表达。最后，与注射CAR-T细胞的小鼠不同，注射7×2b CAR-T细胞的小鼠脾脏和肿瘤组织中CD3表达最为丰富。我们的研究表明，GD2-CAR-T细胞中IL-7和CCR2b共表达比经典第二代CAR-T细胞表现出更强的抗肿瘤活性，为GD2阳性神经母细胞瘤和黑色素瘤潜在的新型治疗方法提供了线索。
