Universal Anti-CD7 CAR-T Cells Targeting T-ALL and Functional Analysis of CD7 Antigen on T/CAR-T Cells.

IF 3.9 3区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Human gene therapy Pub Date : 2023-12-01 Epub Date: 2023-11-24 DOI:10.1089/hum.2023.029
Leling Xie, Runxia Gu, Xue Yang, Shaowei Qiu, Yingxi Xu, Junli Mou, Ying Wang, Haiyan Xing, Kejing Tang, Zheng Tian, Qing Rao, Min Wang, Jianxiang Wang
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引用次数: 0

Abstract

Chimeric antigen receptor T (CAR-T) cell therapy initiates new methods and turns the scale of clinical treatment on relapsed/refractory acute T lymphoblastic leukemia (T-ALL). In this study, we generated the second-generation CD7-targeting CAR-T cells with a new antigen-binding single-chain variable fragment sequence and made it universal via CRISPR-based knockout of TRAC and CD7 genes (termed UCAR-T). The CD7 UCAR-T cells can efficiently proliferate and lyse T-ALL tumor cell in vitro, along with prominent proinflammatory cytokines secretion. A Jurkat-based xenograft mouse model further verified the superior cytotoxicity of the UCAR-T cells in vivo. During the UCAR-T construction, we observed a CD4/CD8 ratio shift among CD7-/- T/CAR-T cells, which motivated us to further analyze the effects of CD7 antigen on T/CAR-T cells. We sorted out CD7+/- T or anti-CD19 CAR-T cells after partially CD7 knockout and performed functional, phenotypic detection, as well as translational analysis. CD7-/- CAR-T cells tended to be CD8 negative and showed slightly better cytotoxicity at long-term assay. RNA-seq further confirmed an elevation of activated CD4 memory cell subpopulation. However, limited distinction on crucial regulatory genes and pathways was revealed, suggesting the safety and feasibility of UCAR-T application as well as the potential translational rather than transcriptional regulation of CD7 antigen.

靶向T-ALL的通用抗CD7 CAR-T细胞和T/CAR-T细胞上CD7抗原的功能分析。
嵌合抗原受体T细胞(CAR-T)疗法开创了复发/难治性急性T淋巴细胞白血病(T-ALL)的新方法,并改变了临床治疗规模。在这项研究中,我们用一种新的抗原结合scFv序列产生了靶向CAR-T细胞的第二代CD7,并通过基于CRISPR的TRAC和CD7基因敲除使其通用(称为UCAR-T)。CD7 UCAR-T细胞可以在体外有效增殖和裂解T-ALL肿瘤细胞,同时分泌显著的促炎细胞因子。基于Jurkat的异种移植物小鼠模型进一步验证了UCAR-T细胞在体内的优越细胞毒性。在UCAR-T构建过程中,我们观察到CD7-/-T/CAR-T细胞中CD4/CD8比例的变化,这促使我们进一步分析CD7抗原对T/CAR-T的影响。我们在部分CD7敲除后筛选出CD7+/-T或抗CD19 CAR-T细胞,并进行功能、表型检测和翻译分析。CD7-/-CAR-T细胞倾向于CD8阴性,并且在长期测定中显示出略好的细胞毒性。RNA-seq进一步证实了活化的CD4记忆细胞亚群的升高。然而,关键调控基因和途径的差异有限,这表明UCAR-T应用的安全性和可行性,以及CD7抗原的潜在翻译而非转录调控。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Human gene therapy
Human gene therapy 医学-生物工程与应用微生物
CiteScore
6.50
自引率
4.80%
发文量
131
审稿时长
4-8 weeks
期刊介绍: Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases.
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