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
Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia.
Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia.
Nature. 2025 Jul 2. doi: 10.1038/s41586-025-09212-7.
The efficacy of chimeric antigen receptor (CAR) T cell therapy in solid tumours is limited by immunosuppression and antigen heterogeneity. To overcome these barriers, 'armoured' CAR T cells, which secrete proinflammatory cytokines, have been developed. However, their clinical application has been limited because of toxicity related to peripheral expression of the armouring transgene. 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.
嵌合抗原受体（CAR）T细胞疗法在实体瘤中的疗效受到免疫抑制和抗原异质性的限制。为克服这些障碍，已开发出分泌促炎细胞因子的“武装”CAR T细胞。然而，由于与武装转基因外周表达相关的毒性，其临床应用受到限制。在此，我们开发了一种CRISPR敲入策略，该策略利用内源基因的调控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基因，我们确定NR4A2和RGS16启动子是有望支持将白细胞介素-12和白细胞介素-2等细胞因子直接递送至肿瘤部位的候选者，从而提高了同基因和异种模型中小鼠的抗肿瘤疗效和长期存活率。这种效应伴随着CAR T细胞多功能性的改善、内源性抗肿瘤免疫的激活以及良好的安全性，并且适用于患者来源的CAR T细胞。
