Feasibility of controlling CD38-CAR T cell activity with a Tet-on inducible CAR design
Feasibility of controlling CD38-CAR T cell activity with a Tet-on inducible CAR design
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DOI：
10.1371/journal.pone.0197349
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发表时间：
2018-05-30
期刊：
影响因子：
3.7
通讯作者：
Mutis, Tuna
中科院分区：
文献类型：
--
作者：
Drent, Esther;Poels, Renee;Mutis, Tuna
嵌合抗原受体（CAR）T细胞近期的临床进展促使CD19 - CARs加速获得临床批准用于治疗急性淋巴细胞白血病。然而，CAR - T细胞疗法存在毒性，尤其是当CARs不完全具有肿瘤特异性时。因此，需要有控制CAR - T细胞活性的策略来提高其安全性。在此，我们以多发性骨髓瘤（MM）相关的CD38分子作为靶分子，测试了一种强力霉素（DOX）诱导的Tet - on CD38 - CAR设计用于控制CAR - T细胞脱靶毒性的可行性和实用性。使用对CD38具有高亲和力的CARs，我们证明该策略能够以依赖DOX剂量的方式适当诱导CD38 - CARs以及CAR介导的T细胞细胞毒性。特别是当DOX剂量限制在10ng/ml时，去除DOX会导致CAR相关的脱肿瘤效应在24小时内相对快速地减弱，这表明对不良CAR活性具有主动可控性。这种Tet - on CAR设计还使我们能够诱导亲和力优化的CD38 - CAR - T细胞发挥最大的抗MM细胞毒性活性，而这些细胞本身已经显示出低毒性特征，从而为这些细胞增加了第二重安全性。总体而言，这些结果表明利用这种DOX诱导的CAR设计来主动调节治疗性T细胞的CAR介导的活性是可能的。因此我们得出结论，与自杀基因相比，Tet - on系统在控制CAR - T细胞的潜在毒性方面可能更具优势，且无需永久性地破坏这些细胞。
Recent clinical advances with chimeric antigen receptor (CAR) T cells have led to the accelerated clinical approval of CD19-CARs to treat acute lymphoblastic leukemia. The CAR T cell therapy is nevertheless associated with toxicities, especially if the CARs are not entirely tumor-specific. Therefore, strategies for controlling the CAR T cell activity are required to improve their safety profile. Here, by using the multiple myeloma (MM)-associated CD38 molecule as target molecule, we tested the feasibility and utility of a doxycycline (DOX) inducible Tet-on CD38-CAR design to control the off-target toxicities of CAR T cells. Using CARs with high affinity to CD38, we demonstrate that this strategy allows the proper induction of CD38-CARs and CAR-mediated T cell cytotoxicity in a DOX-dose dependent manner. Especially when the DOX dose was limited to 10ng/ml, its removal resulted in a relatively rapid decay of CAR-related off-tumor effects within 24 hours, indicating the active controllability of undesired CAR activity. This Tet-on CAR design also allowed us to induce the maximal anti-MM cytotoxic activity of affinity-optimized CD38-CAR T cells, which already display a low toxicity profile, hereby adding a second level of safety to these cells. Collectively, these results indicate the possibility to utilize this DOX inducible CAR-design to actively regulate the CAR-mediated activities of therapeutic T cells. We therefore conclude that the Tet-on system may be more advantageous above suicide-genes to control the potential toxicities of CAR T cells without the need to destroy them permanently.