PiggyBac-Engineered T Cells Expressing CD19-Specific CARs that Lack IgG1 Fc Spacers Have Potent Activity against B-ALL Xenografts
PiggyBac-Engineered T Cells Expressing CD19-Specific CARs that Lack IgG1 Fc Spacers Have Potent Activity against B-ALL Xenografts
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DOI：
10.1016/j.ymthe.2018.05.007
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发表时间：
2018-08-01
影响因子：
12.4
通讯作者：
Micklethwaite, Kenneth P.
中科院分区：
文献类型：
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作者：
Bishop, David C.;Xu, Ning;Micklethwaite, Kenneth P.
CD 19特异性嵌合抗原受体（CAR 19）T细胞的临床试验已证明对复发性和难治性B细胞恶性肿瘤具有显著疗效。与病毒载体相比，piggyBac转座子系统提供了一种不太复杂且更经济的方法来产生CAR 19 T细胞。我们之前已经优化了使用piggyBac系统生成CAR 19 T细胞的方案，但我们发现CAR 19 T细胞的体内功效和持久性较差，这可能是由于有害的Fc γ R与CAR的含IgG 1 Fc间隔区结构域的相互作用。因此，我们设计了三种缺乏IgG 1 Fc区的CD 19特异性汽车，并掺入了CD 28或4-1BB跨膜和共刺激结构域的组合。表达这些重新设计的构建体的PiggyBac产生的CAR 19 T细胞均表现出体外特异性针对CD 19（+）细胞系的反应性。然而，那些结合CD 28跨膜和共刺激结构域显示CD 4优势和较差的细胞毒性。在高剂量下，CAR 19 T细胞在异种移植物小鼠模型中有效对抗B-ALL，无论共刺激结构域如何。在逐渐减少的剂量下，4-1BB共刺激导致CAR 19 T细胞的更大效力和持久性，并提供针对B-ALL再激发的保护。使用piggyBac生产有效的CAR T细胞是简单且具有成本效益的，它可以使CAR T细胞疗法更广泛地获得。
Clinical trials of CD19-specific chimeric antigen receptor (CAR19) T cells have demonstrated remarkable efficacy against relapsed and refractory B cell malignancies. The piggyBac transposon system offers a less complex and more economical means for generating CAR19 T cells compared to viral vectors. We have previously optimized a protocol for the generation of CAR19 T cells using the piggyBac system, but we found that CAR19 T cells had poor in vivo efficacy and persistence, probably due to deleterious Fc gamma R interactions with the CAR's IgG1 Fc-containing spacer domain. We therefore designed three CD19-specifc CARs that lacked the IgG1 Fc region, and we incorporated combinations of CD28 or 4-1BB transmembrane and co-stimulatory domains. PiggyBac-generated CAR19 T cells expressing these re-designed constructs all demonstrated reactivity in vitro specifically against CD19(+) cell lines. However, those combining CD28 transmembrane and co-stimulatory domains showed CD4 predominance and inferior cytotoxicity. At high doses, CAR19 T cells were effective against B-ALL in a xenograft mouse model, regardless of co-stimulatory domain. At diminishing doses, 4-1BB co-stimulation led to greater potency and persistence of CAR19 T cells, and it provided protection against B-ALL re-challenge. Production of potent CAR T cells using piggyBac is simple and cost-effective, and it may enable wider access to CAR T cell therapy.