Optimised modular anti-FLAG CAR T cells for solid tumor therapy

Zhang, X; Xu, R; Zorin, D; Pappas, EG; Tang, J; Bai, Y; Qin, VM; von Scheidt, B; Huang, R; Kulakowska, W; Darido, C; Darcy, PK; Kershaw, MH; Slaney, CY

Optimised modular anti-FLAG CAR T cells for solid tumor therapy

Author(s): Zhang, X; Xu, R; Zorin, D; Pappas, EG; Tang, J; Bai, Y; Qin, VM; von Scheidt, B; Huang, R; Kulakowska, W; Darido, C; Darcy, PK; Kershaw, MH; Slaney, CY;
Details: Publication Year 2025,Volume 14,Issue #8,Page e70046
Journal Title: Clinical & Translational Immunology
Publication Type: Research article
Abstract: OBJECTIVES: Chimeric antigen receptor (CAR) T cell therapies have transformed the treatment of B cell malignancies and show promise in other diseases, including autoimmune disorders and cardiac injury. However, broader application, particularly in solid tumours, is limited by challenges such as antigen escape and tumour heterogeneity. This study aimed to develop an anti-FLAG CAR capable of engaging FLAG-tagged secondary reagents, providing a flexible and adaptable targeting strategy. METHODS: We engineered a humanised anti-FLAG CAR to engage FLAG-tagged secondary reagents. The initial construct exhibited tonic signalling, which was addressed through structural optimisation. Therapeutic efficacy was assessed in solid tumour mouse models expressing either FLAG or a FLAG-tagged secondary targeting reagent. RESULTS: The initial anti-FLAG CAR showed functional activity but exhibited tonic signalling and exhaustion, limiting its therapeutic utility. Structural optimisation significantly reduced exhaustion and improved T cell persistence and functionality. The optimised CAR T cells effectively inhibited tumour growth in models using either FLAG- engineered tumour cells or a FLAG-tagged secondary targeting reagent. CONCLUSION: Our findings underscore the importance of CAR design in minimising exhaustion and enhancing therapeutic efficacy. This work supports a modular CAR T cell platform with the potential to overcome tumour antigen heterogeneity and immune evasion in solid cancers.
Publisher: Wiley
Keywords: Flag; Her2; chimeric antigen receptor; solid cancers; tonic signalling
Department(s): Laboratory Research
Publisher's Version: https://doi.org/10.1002/cti2.70046
Open Access at Publisher's Site: https://doi.org/10.1002/cti2.70046
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2025-09-16 11:58:33

Last Modified: 2025-09-16 11:58:37