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Updated: Jun 7, 2025
A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
Published on: November 12, 2019
Ectopic expression of NKG7 enhances CAR-T function and improves the therapeutic efficacy in liquid and solid tumors
Yuxin Chen1, Meng Wang1, Shuxin Huang2
1Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou, Jiangsu 221004, China; Center of Clinical Oncology, the Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou, Jiangsu 221002, China; Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou, Jiangsu 221004, China.
Abstract:
Lack of biopsies after treatment, especially in solid tumors, restricts the understanding of chimeric antigen receptor (CAR)-T cells -related characteristic in vivo, thus hindering the development of strategies to improve CAR-T cells efficacy. Here, we applied nineteen individual single-cell RNA sequencing (scRNA-seq) data from clinical samples of digestive cancers to explore the characteristics of tumor-infiltrating T cells (TILs) to identify effective targets which might be benefit for enhancing the function of CAR-T cells. The data showed that natural killer cell granule protein 7 (NKG7) was overexpressed in TILs and positively associated with anti-PD1 or anti-CTLA4 therapy in digestive cancers. Subsequently, we found that ectopic expression of NKG7 significantly improved the cytotoxicity of B7H3-targeting CAR-T cells to B7H3-positive digestive cancer cells (MKN45, Huh7, HuCCT-1, SW620 and PANC-1 cells), as well as promoted the TNF-α and IL-2 expression. Furthermore, in a CD19-targeting CAR-T model, the therapeutic efficacy was also found increased after NKG7 overexpression. Mechanically, NKG7 preserved surface CAR expression and promoted CAR-T cell proliferation after exposing to relative tumor antigen. These results indicated that it may be feasible to explore single-cell sequencing data of clinical tumor samples to find strategies to improve CAR-T function, and that ectopic expression of NKG7 is an effective strategy to improve the therapeutic efficacy of CAR-T cells against tumors.
Insights
Natural killer cell granule protein 7 (NKG7) enhances chimeric antigen receptor (CAR)-T cell therapy. Overexpressing NKG7 boosts CAR-T cell effectiveness against digestive cancers by improving cytotoxicity and cytokine production.
Area of Science:
- Immunology
- Oncology
- Genetics
Background:
- Understanding in vivo chimeric antigen receptor (CAR)-T cell characteristics is limited by the lack of post-treatment biopsies, especially in solid tumors.
- This knowledge gap hinders the development of strategies to improve CAR-T cell efficacy.
- Tumor-infiltrating T cells (TILs) play a crucial role in the tumor microenvironment and CAR-T cell function.
Purpose of the Study:
- To explore the characteristics of TILs in digestive cancers using single-cell RNA sequencing (scRNA-seq).
- To identify potential targets for enhancing CAR-T cell function.
- To investigate the role of natural killer cell granule protein 7 (NKG7) in CAR-T cell efficacy.
Main Methods:
- Analysis of nineteen individual scRNA-seq datasets from clinical samples of digestive cancers.
- Investigation of NKG7 expression in TILs and its association with anti-PD1 or anti-CTLA4 therapy.
- Ectopic expression of NKG7 in CAR-T cells targeting B7H3 or CD19.
- Assessment of CAR-T cell cytotoxicity, cytokine production (TNF-α, IL-2), surface CAR expression, and proliferation in vitro and in models.
Main Results:
- NKG7 was found to be overexpressed in TILs and positively associated with anti-PD1/anti-CTLA4 therapy response in digestive cancers.
- Ectopic NKG7 expression significantly enhanced the cytotoxicity of B7H3-targeting CAR-T cells against B7H3-positive digestive cancer cells.
- NKG7 overexpression promoted TNF-α and IL-2 expression in CAR-T cells and improved therapeutic efficacy in a CD19-targeting CAR-T model.
- Mechanistically, NKG7 preserved surface CAR expression and promoted CAR-T cell proliferation upon tumor antigen exposure.
Conclusions:
- Single-cell sequencing of clinical tumor samples is a feasible approach to identify strategies for improving CAR-T cell function.
- Ectopic expression of NKG7 is an effective strategy to enhance the therapeutic efficacy of CAR-T cells against various tumors.
- NKG7 represents a promising target for optimizing CAR-T cell-based cancer immunotherapies.
