Split chimeric antigen receptor-modified T cells targeting glypican-3 suppress hepatocellular carcinoma growth with reduced cytokine release
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Updated: Dec 25, 2025
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
Xuan Liu1, Jianyun Wen1, Honglei Yi2
1Department of Pediatrics, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Background:
Human glypican-3 (hGPC3) is a protein highly expressed in hepatocellular carcinoma (HCC) but limited in normal tissues, making it an ideal target for immunotherapy. The adoptive transfer of hGPC3-specific chimeric antigen receptor T (CAR-T) cells for HCC treatment has been conducted in clinical trials. Due to the rigid construction, conventional CAR-T cells have some intrinsic limitations, like uncontrollable overactivation and inducing severe cytokine release syndrome.
Methods:
We redesigned the hGPC3-specific CAR by splitting the traditional CAR into two parts. By using coculturing assays and a xenograft mouse model, the in vitro and in vivo cytotoxicity and cytokine release of the split anti-hGPC3 CAR-T cells were evaluated against various HCC cell lines and compared with conventional CAR-T cells.
Results:
In vitro data demonstrated that split anti-hGPC3 CAR-T cells could recognize and lyse hGPC3+ HepG2 and Huh7 cells in a dose-dependent manner. Impressively, split anti-hGPC3 CAR-T cells produced and released a significantly lower amount of proinflammatory cytokines, including IFN-γ, TNF-α, IL-6, and GM-CSF, than conventional CAR-T cells. When injected into immunodeficient mice inoculated subcutaneously with HepG2 cells, our split anti-hGPC3 CAR-T cells could suppress HCC tumor growth, but released significantly lower levels of cytokines than conventional CAR-T cells.
Conclusions:
We describe here for the first time the use of split anti-hGPC3 CAR-T cells to treat HCC; split anti-hGPC3 CAR-T cells could suppress tumor growth and reduce cytokine release, and represent a more versatile and safer alternative to conventional CAR-T cells treatment.
Insights
Researchers developed split anti-hGPC3 CAR-T cells to treat hepatocellular carcinoma (HCC). These novel cells effectively suppress tumor growth while significantly reducing harmful cytokine release, offering a safer immunotherapy alternative.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Human glypican-3 (hGPC3) is a promising target for hepatocellular carcinoma (HCC) immunotherapy due to its high expression in tumors and limited presence in normal tissues.
- Conventional chimeric antigen receptor T (CAR-T) cell therapy for HCC shows promise but is limited by potential overactivation and severe cytokine release syndrome.
Purpose of the Study:
- To engineer and evaluate a novel split anti-hGPC3 CAR-T cell therapy for HCC.
- To compare the efficacy and safety profile of split CAR-T cells against conventional CAR-T cells in HCC models.
Main Methods:
- Redesigned the hGPC3-specific CAR by splitting it into two functional parts.
- Assessed in vitro and in vivo cytotoxicity and cytokine release using coculturing assays and a xenograft mouse model.
- Compared split anti-hGPC3 CAR-T cells with conventional CAR-T cells against HCC cell lines.
Main Results:
- Split anti-hGPC3 CAR-T cells demonstrated dose-dependent lysis of hGPC3+ HCC cell lines (HepG2, Huh7) in vitro.
- Significantly lower levels of proinflammatory cytokines (IFN-γ, TNF-α, IL-6, GM-CSF) were released by split CAR-T cells compared to conventional CAR-T cells.
- In vivo, split anti-hGPC3 CAR-T cells suppressed HCC tumor growth in mice with reduced cytokine release.
Conclusions:
- Split anti-hGPC3 CAR-T cells represent a novel and effective approach for HCC treatment.
- This split CAR-T cell design offers a safer alternative to conventional CAR-T cells by mitigating excessive cytokine release.
- Split CAR-T cells show potential as a more versatile and safer immunotherapy for HCC.
