Related Experiment Video
Updated: Jul 14, 2026
Generation of CAR T Cells for Adoptive Therapy in the Context of Glioblastoma Standard of Care
Published on: February 16, 2015
Enhanced anti-tumor activity mediated by combination chimeric antigen receptor T cells targeting GD2 and GPC2 in
Huantong Wu1, Guangji Zhang1, Zhongfeng Liu1
1Cell Therapy Center, Beijing Institute of Geriatrics, Xuanwu Hospital Capital Medical University, National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education, Beijing, China; Center of Neural Injury and Repair, Beijing Institute for Brain Disorders, Beijing, China.
Background Aims:
Chimeric antigen receptor T (CAR-T) cells targeting single antigens show limited activity against solid tumors due to poor T cell persistence, low efficiency infiltration, and exhaustion together with heterogeneous tumor-associated antigen (TAA) expression. This is also true in high-risk neuroblastoma (HRNB), a lethal pediatric extracranial malignancy. To overcome these obstacles, a combinational strategy using GD2-specific and GPC2-specific CAR-T cells was developed to improve immunotherapeutic efficacy.
Methods:
We individually developed GD2-specific and GPC2-specific CARs containing a selective domain (sCAR) which was a peptide of 10 amino acids derived from human nuclear autoantigen La/SS-B. These constructs allowed us to generate two different HRNB antigen-specific CAR-T cells with enhanced biological activity through stimulating sCAR-engrafted T cells via a selective domain-specific monoclonal antibody (SmAb). Binding affinity and stimulation of GD2- and GPC2-specific sCARs by SmAb were measured, and transient and persistent anti-tumor cytotoxicity of GD2sCAR-T and GPC2sCAR-T cells were quantified in neuroblastoma cell lines expressing different TAA levels. The anti-tumor pharmaceutical effects and cellular mechanisms mediated by single or combinational sCAR-T cells were evaluated in vitro and in vivo.
Results:
GD2- and GPC2-specific sCARs had antigen-specific binding affinity similar to their parental counterparts and were recognized by SmAb. SmAb-mediated stimulation selectively activated sCAR-T proliferation and increased central memory T cells in the final products. SmAb-stimulated sCAR-T cells had enhanced transient cytolytic activity, and combination therapy extended long-term anti-tumor activity in vitro through TNF-α and IL-15 release. Stimulated sCAR-T cells overcame heterogeneous antigen expression in HRNB, and the multi-TAA-targeting strategy was especially efficacious in vivo, inducing apoptosis through the caspase-3/PARP pathway and inhibiting the release of several tumor-promoting cytokines.
Conclusions:
These data suggest that combined targeting of multiple TAAs is a promising strategy to overcome heterogenous antigen expression in solid tumors and extend CAR-T cell persistence for HRNB immunotherapy.
Insights
Combining GD2- and GPC2-specific CAR-T cells, activated by a selective monoclonal antibody, enhances anti-tumor activity and persistence for high-risk neuroblastoma immunotherapy.
Area of Science:
- Immunotherapy
- Oncology
- Pediatric Malignancy
Background:
- CAR-T cell therapy faces challenges in solid tumors like high-risk neuroblastoma (HRNB) due to antigen heterogeneity, poor T cell persistence, and exhaustion.
- Current single-antigen CAR-T approaches are insufficient for effective HRNB treatment.
Purpose of the Study:
- To develop a novel combinational CAR-T cell strategy targeting multiple tumor-associated antigens (TAAs) for improved efficacy in HRNB.
- To enhance CAR-T cell biological activity, persistence, and anti-tumor cytotoxicity.
Main Methods:
- Engineered GD2-specific and GPC2-specific chimeric antigen receptors (CARs) with a selective domain (sCAR).
- Stimulated sCAR-T cells using a selective monoclonal antibody (SmAb) to assess proliferation and cytotoxicity in vitro and in vivo.
- Evaluated anti-tumor effects and cellular mechanisms of single and combination sCAR-T cell therapies.
Main Results:
- SmAb-mediated stimulation enhanced sCAR-T cell proliferation and central memory T cell generation.
- Combination therapy demonstrated extended long-term anti-tumor activity and overcame heterogeneous TAA expression in HRNB models.
- In vivo studies showed induced apoptosis via caspase-3/PARP and inhibition of tumor-promoting cytokines.
Conclusions:
- Combined targeting of multiple TAAs with sCAR-T cells is a promising strategy for solid tumor immunotherapy, particularly for HRNB.
- This approach enhances CAR-T cell persistence and overcomes antigen heterogeneity, offering a potential new therapeutic avenue.
More Related Videos
![Dynamic Imaging of Chimeric Antigen Receptor T Cells with [18F]Tetrafluoroborate Positron Emission Tomography/Computed Tomography](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F62334.jpg&w=3840&q=50)
09:34Dynamic Imaging of Chimeric Antigen Receptor T Cells with [18F]Tetrafluoroborate Positron Emission Tomography/Computed Tomography
Published on: February 17, 2022
05:22Intracranial Cannula Implantation for Serial Locoregional Chimeric Antigen Receptor CAR T Cell Infusions in Mice
Published on: February 24, 2023
