Department of Chemistry, Case Western Reserve University, 2080 Adelbert Road, Cleveland, OH 44106, USA.
Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87106, USA.
Int J Mol Sci. 2024 Sep 30;25(19):10568. doi: 10.3390/ijms251910568.
The potential of chimeric antigen receptor (CAR)-based immunotherapy as a promising therapeutic approach is often hindered by the presence of highly immunosuppressive tumor microenvironments (TME). Combination therapies with either co-administration or built-in expression of additional TME-modulating therapeutic molecules to potentiate the functions of CAR-T cells can cause systemic toxicities due to the lack of control over the delivery of biologics. Here, we present a proof-of-concept engineered platform in human Jurkat T cells that combines CAR with a therapeutic gene circuit capable of sensing -galactosidase (a reported cancer-associated signal) and subsequently activate the production of customized therapeutic gene products. We have demonstrated the integration of the chemically induced proximity (CIP) and associated signal sensing technologies with CAR in this study. A -galactosidase-activatable prodrug was designed by conjugating a galactose moiety with a CIP inducer abscisic acid (ABA). We showed that Jurkat T cells engineered with CAR and the ABA-inducible genetic circuits can respond to recombinant -galactosidase to drive the production and secretion of various immunotherapeutics including an anti-cancer agent, an immunomodulatory cytokine, and immune checkpoint inhibitors. Our design is highly modular and could be adapted to sense different cancer-related signals to locally produce antitumor therapeutics that can potentially boost CAR-T efficacy and persistence.
嵌合抗原受体 (CAR) 为基础的免疫疗法作为一种很有前途的治疗方法，其潜力往往受到高度免疫抑制肿瘤微环境 (TME) 的限制。通过联合使用或内置表达其他 TME 调节治疗分子，以增强 CAR-T 细胞的功能，由于缺乏对生物制剂输送的控制，可能会导致全身毒性。在这里，我们提出了一个在人 Jurkat T 细胞中的概念验证工程平台，该平台将 CAR 与一个能够感知β-半乳糖苷酶（一种报道的癌症相关信号）的治疗基因回路结合，随后激活定制治疗基因产物的产生。我们在这项研究中展示了化学诱导接近 (CIP) 及其相关信号传感技术与 CAR 的整合。通过将半乳糖部分与 CIP 诱导剂脱落酸 (ABA) 缀合，设计了一种β-半乳糖苷酶激活的前药。我们表明，工程化的 Jurkat T 细胞与 CAR 和 ABA 可诱导的遗传回路可以响应重组β-半乳糖苷酶，从而驱动各种免疫治疗剂的产生和分泌，包括抗癌剂、免疫调节剂细胞因子和免疫检查点抑制剂。我们的设计具有高度的模块化，可以适应不同的癌症相关信号，以局部产生抗肿瘤治疗剂，这可能会增强 CAR-T 疗效和持久性。
