Rapid generation of a mouse model for evaluating on-target normal tissue toxicity of human CAR-T cells using replication-defective recombinant adenovirus

• Published in: Journal of advanced research 2023-05, Vol.47, p.163-171
• Main Authors: Liao, Qibin, Liu, Zhuoqun, Zhu, Cuisong, He, Huan, Feng, Meiqi, Jiang, Lang, Ding, Xiangqing, Sun, Rongxun, Zhang, Xiaoyan, Xu, Jianqing
• Format: Article
• Language: English
• Subjects: Adenoviridae - genetics; Adenoviridae - metabolism; Adenovirus; Animal model; Animals; CAR-T; CD47 Antigen - metabolism; Humans; Immunotherapy, Adoptive - methods; Mice; Neoplasms - drug therapy; Normal tissue; On-target off-tumor toxicity; Original; Receptors, Chimeric Antigen - genetics; Receptors, Chimeric Antigen - metabolism; T-Lymphocytes; Tumor Microenvironment
• ISSN: 2090-1232; 2090-1224
• DOI: 10.1016/j.jare.2022.08.008

[Display omitted] •Recombinant adenovirus delivers gene into murine tissue to rapidly express human tumor antigen.•Recombinant adenovirus dosage achieves tunable tumor antigen expression in murine tissue.•High levels of tumor antigen in normal tissue results in lethality using CAR-T cells.•Hypoxia-response CAR-T cells ignore normal tissue expressing high levels of tumor antigen.•A mouse model can be used in safety evaluation of CAR-T designs aiming at improving safety in a short time. The on-target off-tumor toxicity of chimeric antigen receptor-engineered T cells (CAR-T) might lead to fatal side effects in cancer patients, which remains as a major obstacle to the clinical application of CAR-T immunotherapy. The off-tumor on-target normal tissue toxicity of CAR-T cells needs to be evaluated in preclinical studies using rational animal models. We aim to develop a rational animal model for assessing the off-tumor on-target normal tissue toxicity of various CAR-T cell designs quickly. We used a recombinant adenovirus type 5 carrying human HER2/ERBB2 (Ad5-HER2) or CD47 gene (Ad5-CD47) to rapidly generate a mouse model with tunable human antigen expression on normal liver tissue to determine immunotoxicity of traditional CAR-T and hypoxia-response CAR-T cells in vivo. The obvious liver damage and lymphocyte infiltration were not observed in mice with human antigen-high livers 8 days post-infection. Interestingly, the lethal liver damage, systemic cytokine release and CAR-T cells infiltration in liver were only observed in mice that received traditional CAR-T cells, but not in hypoxia-response CAR-T cells. Adenovirus-based expression of target antigen in normal mouse tissue may be a useful method for assessing on-target CAR-T cell toxicity in normal tissues, especially various CAR-T cell designs that have the potency of conditional regulation in tumor microenvironment (TME).