Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection.
Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection.
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DOI：
10.1038/nature21405
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发表时间：
2017-03-02
期刊：
影响因子：
64.8
通讯作者：
Sadelain M
中科院分区：
文献类型：
--
作者：
Eyquem J;Mansilla-Soto J;Giavridis T;van der Stegen SJ;Hamieh M;Cunanan KM;Odak A;Gönen M;Sadelain M
嵌合抗原受体(CARS)是一种人工合成的受体，可以重定向和重新编程T细胞，以介导肿瘤排斥反应。到目前为止，最成功的CAR是针对CD19(参考文献)的CAR，它为化疗难治或复发的B细胞恶性肿瘤患者提供了完全缓解的前景。通常使用γ逆转录病毒载体或其他随机整合载体将CARS导入患者的T细胞，这可能导致克隆扩增、致癌转化、多样化的转基因表达和转录沉默。基因组编辑方面的最新进展使人类细胞能够进行有效的序列特异性干预，包括靶向CCR5和AAVS1基因座的基因传递。在这里，我们表明，将CD19特异性CAR导向T细胞受体α常数(TRAC)区域不仅导致人外周血T细胞中CAR的均匀表达，而且还增强了T细胞的效力，在急性淋巴细胞性白血病小鼠模型中，编辑细胞的表现远远优于常规产生的CAR T细胞。我们进一步证明，将CAR靶向TRAC基因座可以避免紧张性CAR信号，并在单次或重复暴露抗原后建立CAR的有效内化和重新表达，延迟效应器T细胞的分化和耗尽。这些发现揭示了CAR免疫生物学的各个方面，并强调了CRISPR/Cas9基因组编辑在推进免疫治疗方面的潜力。
Chimeric antigen receptors (CARs) are synthetic receptors that redirect and reprogram T cells to mediate tumour rejection. The most successful CARs used to date are those targeting CD19 (ref.), which offer the prospect of complete remission in patients with chemorefractory or relapsed B-cell malignancies. CARs are typically transduced into the T cells of a patient using γ-retroviral vectors or other randomly integrating vectors, which may result in clonal expansion, oncogenic transformation, variegated transgene expression and transcriptional silencing. Recent advances in genome editing enable efficient sequence-specific interventions in human cells, including targeted gene delivery to the CCR5 and AAVS1 loci. Here we show that directing a CD19-specific CAR to the T-cell receptor α constant (TRAC) locus not only results in uniform CAR expression in human peripheral blood T cells, but also enhances T-cell potency, with edited cells vastly outperforming conventionally generated CAR T cells in a mouse model of acute lymphoblastic leukaemia. We further demonstrate that targeting the CAR to the TRAC locus averts tonic CAR signalling and establishes effective internalization and re-expression of the CAR following single or repeated exposure to antigen, delaying effector T-cell differentiation and exhaustion. These findings uncover facets of CAR immunobiology and underscore the potential of CRISPR/Cas9 genome editing to advance immunotherapies.