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

Chen, AXY; Yap, KM; Kim, JS; Sek, K; Huang, YK; Dunbar, PA; Wiebking, V; Armitage, JD; Munoz, I; Todd, KL; Derrick, EB; Nguyen, D; Tong, J; Chan, CW; Hoang, TX; Audsley, KM; van Elsas, MJ; Middelburg, J; Lee, JN; de Menezes, MN; Cole, TJ; Li, J; Scheffler, C; Scott, AM; Mackay, LK; Waithman, J; Oliaro, J; Harrison, SJ; Parish, IA; Lai, J; Porteus, MH; House, IG; Darcy, PK; Beavis, PA

Author(s): Chen, AXY; Yap, KM; Kim, JS; Sek, K; Huang, YK; Dunbar, PA; Wiebking, V; Armitage, JD; Munoz, I; Todd, KL; Derrick, EB; Nguyen, D; Tong, J; Chan, CW; Hoang, TX; Audsley, KM; van Elsas, MJ; Middelburg, J; Lee, JN; de Menezes, MN; Cole, TJ; Li, J; Scheffler, C; Scott, AM; Mackay, LK; Waithman, J; Oliaro, J; Harrison, SJ; Parish, IA; Lai, J; Porteus, MH; House, IG; Darcy, PK; Beavis, PA;
Details: Publication Year 2025-08,Volume 644,Issue #8075,Page 241-251
Journal Title: Nature
Publication Type: Research article
Abstract: The efficacy of chimeric antigen receptor (CAR) T cell therapy in solid tumours is limited by immunosuppression and antigen heterogeneity(1-3). To overcome these barriers, 'armoured' CAR T cells, which secrete proinflammatory cytokines, have been developed(4). However, their clinical application has been limited because of toxicity related to peripheral expression of the armouring transgene(5). 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.
Publisher: Springer Nature
Keywords: Animals; Mice; Humans; *Receptors, Chimeric Antigen/genetics/immunology/metabolism; *T-Lymphocytes/immunology/metabolism; Female; *Immunotherapy, Adoptive/methods; *Neoplasms/therapy/immunology/genetics/pathology; Interleukin-2/genetics/immunology/metabolism/administration & dosage; Gene Knock-In Techniques; Promoter Regions, Genetic/genetics; Interleukin-12/genetics/immunology/metabolism; CRISPR-Cas Systems/genetics; Male; Transgenes/genetics; Cell Line, Tumor
Department(s): Laboratory Research; Haematology
Publisher's Version: https://doi.org/10.1038/s41586-025-09212-7
Open Access at Publisher's Site: https://doi.org/10.1038/s41586-025-09212-7
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2025-07-22 03:31:08

Last Modified: 2025-08-28 05:52:25