Gene editing enables T-cell engineering to redirect antigen specificity for potent tumor rejection
Gene editing enables T-cell engineering to redirect antigen specificity for potent tumor rejection
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DOI：
10.26508/lsa.201900367
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发表时间：
2019-04-01
影响因子：
4.4
通讯作者：
Krackhardt, Angela M.
中科院分区：
文献类型：
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作者：
Albers, Julian J.;Ammon, Tim;Krackhardt, Angela M.
TCR转基因T细胞的过继转移在治疗各种癌症方面具有很大的前景。到目前为止，主要采用半随机整合载体对T细胞进行基因修饰。这些具有插入突变的风险，并且外源性TCR的单独添加可能导致TCR链与内源性TCR链错配。使用非病毒载体（如转座子）的现有方法已经降低了插入突变的风险，但尚未实现位点特异性整合。在这里，我们使用CRISPR-Cas9 RNPs和腺相关病毒6进行基因靶向，将工程TCR基因特异性地传递到TCR α恒定位点，从而将其置于内源性转录控制之下。我们的数据表明，这种方法取代了内源性TCR，在体外功能上重定向了编辑过的T细胞的特异性，并在体内异种移植模型中促进了强有力的肿瘤排斥反应。
Adoptive transfer of TCR transgenic T cells holds great promise for treating various cancers. So far, mainly semi-randomly integrating vectors have been used to genetically modify T cells. These carry the risk of insertional mutagenesis, and the sole addition of an exogenous TCR potentially results in the mispairing of TCR chains with endogenous ones. Established approaches using nonviral vectors, such as transposons, already reduce the risk of insertional mutagenesis but have not accomplished site-specific integration. Here, we used CRISPR-Cas9 RNPs and adeno-associated virus 6 for gene targeting to deliver an engineered TCR gene specifically to the TCR alpha constant locus, thus placing it under endogenous transcriptional control. Our data demonstrate that this approach replaces the endogenous TCR, functionally redirects the edited T cells' specificity in vitro, and facilitates potent tumor rejection in an in vivo xenograft model.