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Cited by 195CAR-T cell therapy researchImmunotherapy and Immune ResponsesT-cell and B-cell Immunology

Tuning charge density of chimeric antigen receptor optimizes tonic signaling and CAR-T cell fitness

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Chen, Jian, Qiu, Shizhen, Li, Wentao, Wang, Kun, Zhang, Yu, Yang, Han, Liu, Baichuan, Li, Guangfei +12 more·2023·Cell Research
DOI 10.1038/s41422-023-00789-0PMC PMC10156745
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Abstract

Tonic signaling of chimeric antigen receptor (CAR), i.e., the spontaneous CAR activation in the absence of tumor antigen stimulation, is considered to be a pivotal event controlling CAR-T efficacy. However, the molecular mechanism underlying the spontaneous CAR signals remains elusive. Here, we unveil that positively charged patches (PCPs) on the surface of the CAR antigen-binding domain mediate CAR clustering and result in CAR tonic signaling. For CARs with high tonic signaling (e.g., GD2.CAR and CSPG4.CAR), reducing PCPs on CARs or boosting ionic strength in the culture medium during ex vivo CAR-T cell expansion minimizes spontaneous CAR activation and alleviates CAR-T cell exhaustion. In contrast, introducing PCPs into the CAR with weak tonic signaling, such as CD19.CAR, results in improved in vivo persistence and superior antitumor function. These results demonstrate that CAR tonic signaling is induced and maintained by PCP-mediated CAR clustering. Notably, the mutations we generated to alter the PCPs maintain the antigen-binding affinity and specificity of the CAR. Therefore, our findings suggest that the rational tuning of PCPs to optimize tonic signaling and in vivo fitness of CAR-T cells is a promising design strategy for the next-generation CAR.

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Cited by

Sorted by citations

Authors
Chen, J., Qiu, S., Li, W., Wang, K., Zhang, Y., Yang, H., Liu, B., Li, G., Li, L., Chen, M., Lan, J., Niu, J., He, P., Cheng, L., Fan, G., Liu, X., Song, X., Xu, C., Wu, H., & Wang, H.
Affiliations
  • Fudan University (CN)
  • ShanghaiTech University (CN)
  • Austrian Academy of Sciences (AT)
  • Chinese Academy of Sciences (CN)
  • Shanghai Jiao Tong University (CN)
  • Shanghai Medical College of Fudan University (CN)
Corresponding author
  • Song, Xianmin
  • Xu, Chenqi
  • Wu, Haitao
  • Wang, Haopeng
ORCID
Venue
Cell Research
Publisher
Springer Science and Business Media LLC
Published
2023-03-08
Volume
33
Issue
5
Pages
341-354
ISSN
  • 1001-0602 (ISSN-L)
  • 1748-7838
DOI
10.1038/s41422-023-00789-0
PMC
PMC10156745
OpenAlex
W4323349483
Cited by
195
References
4
Citations by year
2026: 62 · 2025: 58 · 2024: 57 · 2023: 18
FWCI
23.79
Type
journal article
Language
en
Update notices
correction (2026-09-14)
Open access
Licence
Locations
  • Cell Research (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Youth Foundation of China (82102863)
  • Shanghai Science and Technology Development Foundation (21ZR1480200, 21Y11912000)
  • National Natural Science Foundation of China (82271149)
Topics
  • Health Sciences › Medicine › Oncology › CAR-T cell therapy research
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Keywords
chimeric antigen receptor, tonic signaling, CAR-T cell exhaustion, antigen-binding domain, CD19 CAR, ionic strength, CAR T cell persistence, automotive design
MeSH terms
Receptors, Chimeric Antigen, Antigens, Neoplasm, T-Lymphocytes, Signal Transduction, Immunotherapy, Adoptive

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