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构建 PD1/CD28 嵌合开关受体增强 c-Met CAR-T 在胃癌中的抗肿瘤能力。

Construction of PD1/CD28 chimeric-switch receptor enhances anti-tumor ability of c-Met CAR-T in gastric cancer.

机构信息

Lanzhou University Second Hospital, the Second Clinical Medical College of Lanzhou University, Lanzhou, Gansu, China.

Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.

出版信息

Oncoimmunology. 2021 Mar 31;10(1):1901434. doi: 10.1080/2162402X.2021.1901434.

Abstract

Chimeric antigen receptor (CAR) T cell is a promising method in cancer immunotherapy but faces many challenges in solid tumors. One of the major problems was immunosuppression caused by PD-1. In our study, the expression of c-Met in GC was analyzed from TCGA datasets, GC tissues, and cell lines. The c-Met CAR was a second-generation CAR with 4-1BB, cMet-PD1/CD28 CAR was c-Met CAR adding PD1/CD28 chimeric-switch receptor (CSR). In , we measured the changes of different subgroups, phenotypes and PD-1 expression in CAR-T cells. We detected the secretion levels of different cytokines and the killing ability of CAR-Ts. In , we established a xenograft GC model and observed the anti-tumor effect and off-target toxicity of different CAR-Ts. We find that the expression of c-Met was increased in GC. CD3CD8 T cells and CD62LCCR7 central memory T cells (T) were increased in two CAR-Ts. The stimulation of target cells could promote the expression of PD-1 in c-Met CAR-T. Compared with Mock T, the secretion of cytokines as IFN-γ, TNF-α, IL-6, IL-10 secreted by two CAR-Ts was increased, and the killing ability to c-Met positive GC cells was enhanced. The PD1/CD28 CSR could further enhance the killing ability, especially the long-term anti-tumor effect of c-Met CAR-T, and reduce the release level of IL-6. CAR-Ts target c-Met had no obvious off-target toxicity to normal organs. Thus, the PD1/CD28 CSR could further enhance the anti-tumor ability of c-Met CAR-T, and provides a promising design strategy to improve the efficacy of CAR-T in GC.

摘要

嵌合抗原受体 (CAR) T 细胞在癌症免疫治疗中是一种很有前途的方法,但在实体瘤中面临许多挑战。其中一个主要问题是由 PD-1 引起的免疫抑制。在我们的研究中,我们从 TCGA 数据集、GC 组织和细胞系中分析了 GC 中 c-Met 的表达。c-Met CAR 是一种第二代 CAR,具有 4-1BB,cMet-PD1/CD28 CAR 是在 c-Met CAR 中添加 PD1/CD28 嵌合开关受体 (CSR)。在 ,我们测量了不同亚组、表型和 CAR-T 细胞中 PD-1 表达的变化。我们检测了不同细胞因子的分泌水平和 CAR-T 的杀伤能力。在 ,我们建立了异种移植 GC 模型,观察了不同 CAR-T 的抗肿瘤作用和脱靶毒性。我们发现 GC 中 c-Met 的表达增加。两种 CAR-T 中 CD3CD8 T 细胞和 CD62LCCR7 中央记忆 T 细胞 (T) 增加。靶细胞的刺激可以促进 c-Met CAR-T 中 PD-1 的表达。与 Mock T 相比,两种 CAR-T 分泌的细胞因子如 IFN-γ、TNF-α、IL-6、IL-10 增加,对 c-Met 阳性 GC 细胞的杀伤能力增强。PD1/CD28 CSR 可以进一步增强杀伤能力,特别是 c-Met CAR-T 的长期抗肿瘤作用,并降低 IL-6 的释放水平。CAR-T 靶向 c-Met 对正常器官没有明显的脱靶毒性。因此,PD1/CD28 CSR 可以进一步增强 c-Met CAR-T 的抗肿瘤能力,为提高 CAR-T 在 GC 中的疗效提供了一种有前途的设计策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f59f/8018404/3caf0af21f5b/KONI_A_1901434_F0001_OC.jpg

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