Moderate expression of CD39 in GPC3-CAR-T cells shows high efficacy against hepatocellular carcinoma

Abstract

CD39 serves as a crucial biomarker for neoantigen-specific CD8⁺ T cells and is associated with antitumor activity and exhaustion. However, the relationship between CD39 expression levels and the function of chimeric antigen receptor T (CAR-T) cells remains controversial. This study aimed to investigate the role of CD39 in the functional performance of CAR-T cells against hepatocellular carcinoma (HCC) and explore the therapeutic potential of CD39 modulators, such as mitochondrial division inhibitor-1 (mdivi-1), or knockdown CD39 through short hairpin RNA. Our findings demonstrated that glypican-3-CAR-T cells with moderate CD39 expression exhibited a strong antitumor activity, while high and low levels of CD39 led to an impaired cellular function. Methods modulating the proportion of CD39 intermediate (CD39^int)-phenotype CAR-T cells such as mdivi-1 and CD39 knockdown enhanced and impaired T cell function, respectively. The combination of mdivi-1 and CD39 knockdown in CAR-T cells yielded the highest proportion of infiltrated CD39^int CAR-T cells and demonstrated a robust antitumor activity in vivo. In conclusion, this study revealed the crucial role of CD39 in CAR-T cell function, demonstrated the potential therapeutic efficacy of combining mdivi-1 with CD39 knockdown in HCC, and provided a novel treatment strategy for HCC patients in the field of cellular immunotherapy.

Keywords

CD39 / CAR-T cells / mdivi-1 / hepatocellular carcinoma / antitumor activity

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (Nos. 82102169, 82003252, 82202986, and 82301960), Outstanding Young Talents Seedling Program of Guangdong Hospital of Traditional Chinese Medicine (No. SZ2023QN03), Young Doctor “Sailing” Project of Science and Technology Department of Guangzhou (No. 2024A04J3291), and Shenzhen Municipal Government of China (No. JCYJ20210324102807019).

Electronic Supplementary Material

Supplementary material is available in the online version of this article at https://doi.org/10.1007/s11684-024-1071-9 and is accessible for authorized users.

Compliance with ethics guidelines

Conflicts of interest Fan Zou, Jialiang Wei, Jialang Zhuang, Yafang Liu, Jizhou Tan, Xianzhang Huang, and Ting Liu have declare that they have no conflict of interest.

The study was approved by the ethics committee of the First Affiliated Hospital of Sun Yat-sen University, under permit number 2018 [43] and the study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. Informed consent was obtained from all patients for being included in the study.

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