Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma
Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma
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DOI：
10.1038/mt.2015.71
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发表时间：
2015-08-01
影响因子：
12.4
通讯作者：
Feldman, Steven A.
中科院分区：
文献类型：
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作者：
Beane, Joal D.;Lee, Gary;Feldman, Steven A.
程序性细胞死亡-1（PD-1）在活化的T细胞上表达，代表了过继性细胞转移（ACT）之前肿瘤靶向T细胞基因编辑的有吸引力的靶标。我们使用针对编码人PD-1（PDCD-1）的基因的锌指核酸酶（ZFN）对黑色素瘤肿瘤浸润淋巴细胞（TIL）进行基因编辑。我们表明，我们的临床规模的TIL生产过程产生了PD-1基因座的有效修饰，在大量TIL群体中等位基因的平均修饰频率为74.8%（n = 3，范围69.9-84.1%），这导致PD-1表面表达降低76%。40%至48%的PD-1基因编辑细胞具有双等位基因PD-1修饰。重要的是，在测试的三个供体中的两个中，与未修饰的TIL相比，PD-1基因编辑的TIL产物显示出改善的体外效应子功能和显著增加的多功能细胞因子谱（TNF α、GM-CSF和IFN γ）。此外，所有供体细胞均显示效应记忆表型，并在体外扩增约500- 2，000倍。因此，需要进一步研究以确定使用PD-1基因编辑的TIL治疗转移性黑色素瘤的过继细胞转移的效率和安全性。
Programmed cell death-1 (PD-1) is expressed on activated T cells and represents an attractive target for gene-editing of tumor targeted T cells prior to adoptive cell transfer (ACT). We used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma tumor infiltrating lymphocytes (TIL). We show that our clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% (n = 3, range 69.9-84.1%) of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression. Forty to 48% of PD-1 gene-edited cells had biallelic PD-1 modification. Importantly, the PD-1 gene-edited TIL product showed improved in vitro effector function and a significantly increased polyfunctional cytokine profile (TNF alpha, GM-CSF, and IFN gamma) compared to unmodified TIL in two of the three donors tested. In addition, all donor cells displayed an effector memory phenotype and expanded approximately 500-2,000-fold in vitro. Thus, further study to determine the efficiency and safety of adoptive cell transfer using PD-1 gene-edited TIL for the treatment of metastatic melanoma is warranted.