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GPC3-targeted CAR-T cells expressing GLUT1 or AGK exhibit enhanced antitumor activity against hepatocellular carcinoma

Acta Pharmacologica Sinica (2024)Cite this article

Abstract

Chimeric antigen receptor-expressing T (CAR-T) cells induce robust antitumor responses in patients with hematologic malignancies. However, CAR-T cells exhibit only limited efficacy against solid tumors such as hepatocellular carcinoma (HCC), partially due to their limited expansion and persistence. CD8+ T cells, as key components of the adaptive immune response, play a central role in antitumor immunity. Aerobic glycolysis is the main metabolic feature of activated CD8+ T cells. In the tumor microenvironment, however, the uptake of large amounts of glucose by tumor cells and other immunosuppressive cells can impair the activation of T cells. Only when tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment have a glycolytic advantage might the effector function of T cells be activated. Glucose transporter type 1 (GLUT1) and acylglycerol kinase (AGK) can boost glycolytic metabolism and activate the effector function of CD8+ T cells, respectively. In this study, we generated GPC3-targeted CAR-T cells overexpressing GLUT1 or AGK for the treatment of HCC. GPC3-targeted CAR-T cells overexpressing GLUT1 or AGK specifically and effectively lysed GPC3-positive tumor cells in vitro in an antigen-dependent manner. Furthermore, GLUT1 or AGK overexpression protected CAR-T cells from apoptosis during repeated exposures to tumor cells. Compared with second-generation CAR-T cells, GPC3-targeted CAR-T cells overexpressing GLUT1 or AGK exhibited greater CD8+ T-cell persistence in vivo and better antitumor effects in HCC allograft mouse models. Finally, we revealed that GLUT1 or AGK maintained anti-apoptosis ability in CD8+ T cells via activation of the PI3K/Akt pathway. This finding might identify a therapeutic strategy for advanced HCC.

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Fig. 1: Increased glycolysis improves CD8+ T cell function.
Fig. 2: Generation of GPC3-targeted CAR-T cells overexpressing GLUT1 or AGK.
Fig. 3: Immune function of GLUT1 and AGK CAR-T cells.
Fig. 4: Reduced apoptosis in CD8+ T cells expressing the GLUT1 or AGK CAR.
Fig. 5: GLUT1- or AGK-overexpressing CAR-T cells displayed superior antitumor efficacy in vivo.
Fig. 6: Accumulation of CD8+ T cells after effective dual-targeted CAR-T cell therapy in vivo.
Fig. 7: GLUT1 and AGK maintain apoptosis resistance in CD8+ T cells via the PI3K/Akt pathway.

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Data availability

The human sequence data generated in this study are not publicly available due to privacy requirements but are available upon reasonable request from the corresponding author. Other data generated in this study are available within the article and its supplementary data files.

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Acknowledgements

Funding

This study was supported by the National Natural Science Foundation of China (ID 82073358 to Hua Jiang, ID 82073359 to Zonghai Li, 82272920 to Min Zhou), the Shanghai Municipal Health and Health Commission (ID 20234Y0193 to Sun Ruixin), the Shanghai Oriental Talent Youth Project (to Sun Ruixin), the Shanghai “Rising Stars of Medical Talents” Youth Development Program-Clinical Laboratory Practitioner Program (ID 2023-JY to Sun Ruixin) and the “Gan Quan Xin Xing” talent training program of Shanghai Tongji Hospital (HRBC 2307).

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Authors and Affiliations

  1. State Key Laboratory of Systems Medicine for Cancer, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200032, China

    Rui-xin Sun, Yi-fan Liu, Yan-sha Sun, Min Zhou, Yi Wang, Bi-zhi Shi, Hua Jiang & Zong-hai Li

  2. Department of Laboratory Medicine, Shanghai Tongji Hospital, School of Medicine, Tongji University, Shanghai, 200065, China

    Rui-xin Sun

  3. CARsgen Therapeutics, Shanghai, 200032, China

    Yi Wang, Bi-zhi Shi, Hua Jiang & Zong-hai Li

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Contributions

SRX, JH, and LZH designed the study. RXS, LYF, SYS, WY, and SBZ performed the experiments and analyzed the data. SRX and LYF performed the animal experiments. SRX wrote the paper. JH and LZH reviewed and edited the manuscript. SRX, JH, ZM, and LZH supervised the project.

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Correspondence to Hua Jiang or Zong-hai Li.

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Competing interests

Dr. Zonghai Li has ownership interests in CAR-T cells related to this work. The remaining authors declare no competing interests.

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The Ethics Committee of Renji Hospital, Shanghai Jiaotong University School of Medicine (RJ2022-1115) approved this study. All animal experiments were performed according to protocols approved by the Shanghai Cancer Institute Experimental Animal Care Commission.

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Sun, Rx., Liu, Yf., Sun, Ys. et al. GPC3-targeted CAR-T cells expressing GLUT1 or AGK exhibit enhanced antitumor activity against hepatocellular carcinoma. Acta Pharmacol Sin (2024). https://doi.org/10.1038/s41401-024-01287-8

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