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Updated: Aug 31, 2025
Author Spotlight: Enhancing CAR-T Cell Function in Syngeneic Tumor Models
Published on: February 2, 2024
Rapid generation of a mouse model for evaluating on-target normal tissue toxicity of human CAR-T cells using
Qibin Liao1, Zhuoqun Liu2, Cuisong Zhu2
1Zhongshan Hospital, Institutes of Biomedical Sciences and Shanghai Public Health Clinical Center, Fudan University, Shanghai, China; Department of Oncology and Bio-therapeutic Center, Shenzhen Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen, China.
Introduction:
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.
Objectives:
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.
Methods:
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.
Results:
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.
Conclusion:
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).
Insights
A new mouse model using adenovirus allows rapid assessment of CAR-T cell toxicity in normal tissues. Hypoxia-response CAR-T cells showed reduced liver damage compared to traditional CAR-T cells, improving immunotherapy safety.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Chimeric antigen receptor-engineered T cell (CAR-T) therapy faces challenges due to on-target, off-tumor toxicity, limiting clinical application.
- Evaluating normal tissue toxicity of CAR-T cells requires effective preclinical animal models.
Purpose of the Study:
- To develop a rapid, rational animal model for assessing off-tumor, on-target normal tissue toxicity of diverse CAR-T cell designs.
- To evaluate the in vivo immunotoxicity of traditional and hypoxia-response CAR-T cells.
Main Methods:
- Utilized recombinant adenovirus type 5 (Ad5-HER2 or Ad5-CD47) to create a mouse model with tunable human antigen expression in normal liver tissue.
- Assessed toxicity of traditional CAR-T and hypoxia-response CAR-T cells in the generated mouse model.
Main Results:
- No significant liver damage or lymphocyte infiltration observed in mice with high antigen expression 8 days post-infection.
- Lethal liver damage, cytokine release, and CAR-T cell infiltration occurred only with traditional CAR-T cells, not hypoxia-response CAR-T cells.
Conclusions:
- Adenovirus-mediated antigen expression in normal tissue provides a valuable method for evaluating on-target CAR-T cell toxicity.
- This model is particularly useful for assessing CAR-T cell designs with conditional regulation capabilities in the tumor microenvironment (TME).
