Lainšček Duško, Golob-Urbanc Anja, Mikolič Veronika, Pantović-Žalig Jelica, Malenšek Špela, Jerala Roman
Department of Synthetic Biology and Immunology, National Institute of Chemistry, Hajdrihova 19, Ljubljana 1000, Slovenia.
EN-FIST Centre of Excellence, Trg Osvobodilne fronte 13, Ljubljana 1000, Slovenia.
Mol Ther Oncolytics. 2023 Apr 26;29:77-90. doi: 10.1016/j.omto.2023.04.005. eCollection 2023 Jun 15.
CAR-T cells present a highly effective therapeutic option for several malignant diseases, based on their ability to recognize the selected tumor surface marker in an MHC-independent manner. This triggers cell activation and cytokine production, resulting in the killing of the cancerous cell presenting markers recognized by the chimeric antigen receptor. CAR-T cells are highly potent serial killers that may cause serious side effects, so their activity needs to be carefully controlled. Here we designed a system to control the proliferation and activation state of CARs based on downstream NFAT transcription factors, whose activity can be regulated via chemically induced heterodimerization systems. Chemical regulators were used to either transiently trigger engineered T cell proliferation or suppress CAR-mediated activation when desired or to enhance activation of CAR-T cells upon engagement of cancer cells, shown also . Additionally, an efficient sensor to monitor activated CD19 CAR-T cells was introduced. This implementation in CAR-T cell regulation offers an efficient way for on-demand external control of CAR-T cell activity to improve their safety.
嵌合抗原受体（CAR）修饰的T细胞（CAR-T细胞）对多种恶性疾病而言是一种高效的治疗选择，这是基于它们能够以不依赖主要组织相容性复合体（MHC）的方式识别所选肿瘤表面标志物。这会触发细胞活化和细胞因子产生，从而导致表达被嵌合抗原受体识别的标志物的癌细胞被杀伤。CAR-T细胞是高效的连续杀手，可能会引起严重的副作用，因此需要仔细控制它们的活性。在此，我们设计了一个基于下游活化T细胞核因子（NFAT）转录因子来控制CAR增殖和活化状态的系统，其活性可通过化学诱导异二聚化系统进行调节。化学调节剂用于在需要时短暂触发工程化T细胞增殖或抑制CAR介导的活化，或者在癌细胞接触时增强CAR-T细胞的活化，这也已得到证实。此外，还引入了一种用于监测活化的CD19 CAR-T细胞的高效传感器。这种在CAR-T细胞调控中的应用为按需外部控制CAR-T细胞活性以提高其安全性提供了一种有效方法。
