We elucidate the therapeutic benefits of CAR T cells in HCC and identify the primary barriers to their broader application. Our analysis aims to provide a comprehensive overview of the current status and future prospects of CAR T-cell immunotherapy for HCC.

P1, N=15, Recruiting, Zhejiang University | Not yet recruiting --> Recruiting

However, in advanced HCC, clinical trials have not demonstrated sufficient anti-tumor efficacy, in contrast with preclinical studies. Reverse translation, bedside-to-bench research, is crucial to identify the factors that have hindered GPC3 target immunotherapies.

P1/2, N=30, Not yet recruiting, Chinese PLA General Hospital

GPC-3's re-expression in HCC and its involvement in key tumorigenic processes underscore its value as a biomarker for early diagnosis and a target for therapeutic intervention. Further research is warranted to fully exploit GPC-3's diagnostic and therapeutic potential in HCC management.

The protocol includes producing retroviral vectors, generating CAR T cells, and conducting ecto-ADA1 activity, cytotoxicity, cell migration, and RNA sequencing assays. For complete details on the use and execution of this protocol, please refer to Hu et al.1.

P=N/A, N=12, Not yet recruiting, The First People's Hospital of Lianyungang; The First People's Hospital of Lianyungang | Initiation date: May 2024 --> Sep 2024

P1/2, N=121, Not yet recruiting, Shanghai AbelZeta Ltd.

Overall, our study successfully generated GPC3 CAR-T cells expressing both IL-21 and CXCL9, demonstrated that combining PD-1 blockade can further enhance CAR-T cell function by promoting proliferation, cytokine secretion, chemotaxis and antitumor activity. These findings present a hopeful and potentially effective strategy for GPC3-positive HCC patients.

P1, N=24, Recruiting, Baylor College of Medicine | Not yet recruiting --> Recruiting

P1, N=30, Recruiting, CARsgen Therapeutics Co., Ltd.

P1, N=10, Terminated, jianming xu | N=34 --> 10 | Trial completion date: Oct 2026 --> Jul 2024 | Recruiting --> Terminated; achieve the proof of concept

P1, N=11, Active, not recruiting, Takeda | Trial completion date: Dec 2026 --> Jul 2026 | Trial primary completion date: Dec 2026 --> Jul 2026

This study provides a rationale for NT-I7 plus CAR-T cell combination therapy for solid tumors in humans.

P=N/A, N=12, Recruiting, The First People's Hospital of Lianyungang; The First People's Hospital of Lianyungang

P=N/A, N=64, Not yet recruiting, AstraZeneca | Initiation date: Jun 2024 --> Oct 2024

P1, N=30, Recruiting, Second Affiliated Hospital of Guangzhou Medical University | Not yet recruiting --> Recruiting | Trial completion date: Dec 2033 --> Dec 2036 | Initiation date: Jan 2024 --> May 2024 | Trial primary completion date: Nov 2026 --> Nov 2030

P1, N=30, Recruiting, Second Affiliated Hospital of Guangzhou Medical University | Trial completion date: Aug 2026 --> Aug 2036 | Trial primary completion date: Aug 2024 --> Aug 2026

P1, N=30, Recruiting, Second Affiliated Hospital of Guangzhou Medical University | Trial completion date: Mar 2033 --> Mar 2036 | Trial primary completion date: Mar 2026 --> Mar 2029

P1, N=30, Recruiting, Second Affiliated Hospital of Guangzhou Medical University | Trial completion date: Aug 2029 --> Aug 2036 | Trial primary completion date: Aug 2025 --> Aug 2029

The anti-FAP-GPC3 bispecific CAR-T cells also exhibited superior antitumor efficacy and significantly prolonged the survival of mice compared with single-target CAR-T cells in vivo. Overall, the use of anti-FAP-GPC3 bispecific CAR-T cells is a promising treatment approach to reduce tumor recurrence caused by tumor antigen heterogeneity.

In 3D NACs-origami spheroid model of HCC, CAR-Ms were further demonstrated to have a significant tumor killing effect. Together, our study provides a new strategy for the treatment of HCC through CAR-M cells targeting GPC3, which provides a basis for the research and treatment of hepatocellular carcinoma.

P1, N=15, Not yet recruiting, Zhejiang University

The combination of mdivi-1 and CD39 knockdown in CAR-T cells yielded the highest proportion of infiltrated CD39int 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.

In this study, lapatinib-resistant cells were used to test the concept of "Evolutionary Traps" and no suitable target stand out because of the identified genes without accessible drug...To test whether this concept is cell line or drug dependent, we analyzed 21 drug-resistant tumor cell expression profiles reveal that JAG1, GPC3, and L1CAM, which are suitable targets for CAR-T treatment, are significantly upregulated in various drug-resistant tumor cells. Our findings shed light on the feasibility of utilizing CAR-T therapy to treat drug-resistant tumors and broaden the concept of the "Evolutionary Trap".

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.

P1, N=9, Not yet recruiting, Beijing Immunochina Medical Science & Technology Co., Ltd.

P=N/A, N=64, Not yet recruiting, AstraZeneca | N=30 --> 64 | Initiation date: Feb 2024 --> Jun 2024

P1, N=12, Recruiting, Eutilex | Not yet recruiting --> Recruiting | Trial completion date: Nov 2024 --> Dec 2025 | Trial primary completion date: May 2024 --> Jun 2025

P1, N=48, Recruiting, Zhejiang University | N=20 --> 48

P1, N=21, Not yet recruiting, Baylor College of Medicine

P1/2, N=110, Recruiting, Sotio Biotech Inc. | Trial completion date: Jul 2039 --> Dec 2041 | Trial primary completion date: Jul 2024 --> Apr 2026

P=N/A, N=30, Not yet recruiting, AstraZeneca

P1, N=11, Active, not recruiting, Takeda | Recruiting --> Active, not recruiting | N=18 --> 11

P1, N=3, Active, not recruiting, Tongji University | Recruiting --> Active, not recruiting | Trial completion date: Sep 2022 --> Sep 2024 | Trial primary completion date: Aug 2022 --> Aug 2024

P1/2, N=84, Recruiting, AstraZeneca | Not yet recruiting --> Recruiting

In conclusion, GPC3-7-19-CAR-T cells achieved antitumor effects superior to those of conventional GPC3-CAR-T cells by reconstructing the TME induced by the dominant CD4 T and CD8 T cell subsets. Most importantly, GPC3-7-19-CAR-T cells exhibited good safety and antitumor efficacy in HCC patients in the clinic. ► Novel GPC3-7-19-CAR-T cells designed with mediate level of IL-7 secretion and high level of CCL19 secretion, which could recruit more mature DCs to assist killing on GPC3HCCs. ►DC cells recruited by CCL19 could interact with CD4 T cells and promote the differentiation of CD4T cells into CD4T and CD8T subsets, leading a better anti-tumor effect on GPC3HCCs. ►Compared with conventional GPC3-CAR-T, GPC3-7-CCL19-CAR-T cells could reverse tumor immunosuppressive microenvironment by reducing PMN-MDSC and Treg cell infiltration.

P1, N=20, Recruiting, Shanghai Ming Ju Biotechnology Co., Ltd.

P1/2, N=84, Not yet recruiting, AstraZeneca

GPC3 protein is expressed in a large percentage of HB and FLC tumors, but with variable intensity and distribution. Characterization of GPC3 tumor protein H-score and pattern of immunoreactivity are two independent parameters that may be helpful to quantitate GPC3 immunostaining of tumor and correlate with treatment response to a variety of GPC3 targeted therapeutics.

Importantly, STK-009 induced the expression of cytotoxic machinery, pro-survival, and proliferative genes in tumor-infiltrating SYNCAR-002 (figure 3). Conclusions These findings validate that the orthogonal IL-2 platform has the potential to improve the efficacy and durability of CAR T therapy for solid tumor targets such as GPC3 by selectively expanding CAR-T cells in vivo and activating CAR-T cells in a hostile tumor microenvironment.

CT017 CAR T cells were engineered to co-express CAR-GPC3 and runt-related transcription factor 3 (RUNX3), which triggers CD8 T-cell infiltration into the cancer microenvironment...These results need to be confirmed in a robust clinical trial. This study was funded by CARsgen Therapeutics Co., Ltd.