墨尔本大学Paul A. Beavis团队近日取得一项新成果。经过不懈努力，他们揭示了在CAR-T细胞中重新连接内源性基因用于肿瘤限制性有效载荷递送。相关论文于2025年7月2日发表在《自然》杂志上。

该研究团队已经开发了一种CRISPR敲入策略，利用内源性基因的调节机制，以肿瘤定位的方式驱动转基因表达。通过筛选具有肿瘤限制性表达的内源性基因，该研究团队已经确定NR4A2和RGS16启动子是支持细胞因子如IL-12和IL-2直接传递到肿瘤部位的有希望的候选基因，从而提高了抗肿瘤功效和小鼠的长期存活，无论是在同种还是异种模型中。这种效果伴随着CAR-T细胞多功能性的改善、内源性抗肿瘤免疫的激活和良好的安全性，并且适用于来自患者的CAR-T细胞。

据悉，嵌合抗原受体（CAR） T细胞治疗实体肿瘤的疗效受到免疫抑制和抗原异质性的限制。为了克服这些障碍，分泌促炎细胞因子的“武装”CAR-T细胞被开发出来。然而，它们的临床应用一直受到限制，因为它们的毒性与手臂转基因的外周表达有关。

附：英文原文

Title: Rewiring endogenous genes in CAR T cells for tumour-restricted payload delivery

Author: Chen, Amanda X. Y., Yap, Kah Min, Kim, Joelle S., Sek, Kevin, Huang, Yu-Kuan, Dunbar, Phoebe A., Wiebking, Volker, Armitage, Jesse D., Munoz, Isabelle, Todd, Kirsten L., Derrick, Emily B., Nguyen, Dat, Tong, Junming, Chan, Cheok Weng, Hoang, Thang X., Audsley, Katherine M., van Elsas, Marit J., Middelburg, Jim, Lee, Joel N., de Menezes, Maria N., Cole, Thomas J., Li, Jasmine, Scheffler, Christina, Scott, Andrew M., Mackay, Laura K., Waithman, Jason, Oliaro, Jane, Harrison, Simon J., Parish, Ian A., Lai, Junyun, Porteus, Matthew H., House, Imran G., Darcy, Phillip K., Beavis, Paul A.

Issue&Volume: 2025-07-02

Abstract: The efficacy of chimeric antigen receptor (CAR) T cell therapy in solid tumours is limited by immunosuppression and antigen heterogeneity1,2,3. To overcome these barriers, ‘armoured’ CAR T cells, which secrete proinflammatory cytokines, have been developed4. However, their clinical application has been limited because of toxicity related to peripheral expression of the armouring transgene5. Here, we have developed a CRISPR knock-in strategy that leverages the regulatory mechanisms of endogenous genes to drive transgene expression in a tumour-localized manner. By screening endogenous genes with tumour-restricted expression, we have identified the NR4A2 and RGS16 promoters as promising candidates to support the delivery of cytokines such as IL-12 and IL-2 directly to the tumour site, leading to enhanced antitumour efficacy and long-term survival of mice in both syngeneic and xenogeneic models. This effect was concomitant with improved CAR T cell polyfunctionality, activation of endogenous antitumour immunity and a favourable safety profile, and was applicable in CAR T cells from patients.

DOI: 10.1038/s41586-025-09212-7

Source: https://www.nature.com/articles/s41586-025-09212-7