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Updated: Aug 1, 2025
Generation of Human Chimeric Antigen Receptor Regulatory T Cells
Published on: January 3, 2025
Design of diversified chimeric antigen receptors through rational module recombination
Wen Si1,2, Ying-Ying Fan1,2, Shi-Zhen Qiu3
1Center for Cell and Gene Circuit Design, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
Abstract:
Chimeric antigen receptor (CAR)-T cells have shown great promise in cancer therapy. However, the anti-tumor efficiency is limited due to the CAR-induced T cell apoptosis or exhaustion. The intracellular domain of CAR comprised of various signaling modules orchestrates CAR-T cell behaviors. The modularity of CAR signaling domain functions as the "mainboard" to assemble diversified downstream signaling components. Here, we implemented the modular recombination strategy to construct a library of CARs with synthetic co-signaling modules adopted from immunoglobin-like superfamily (IgSF) and tumor necrosis factor receptor superfamily (TNFRSF). We quantitatively characterized the signaling behaviors of these recombinants by both NFAT and NF-κB reporter, and identified a set of new CARs with diverse signaling behaviors. Specifically, the 28(NM)-BB(MC) CAR-T cells exhibited improved cytotoxicity and T cell persistence. The synthetic approach can promote our understanding of the signaling principles of CAR molecule, and provide a powerful tool box for CAR-T cell engineering.
Insights
Engineered chimeric antigen receptor (CAR)-T cells show promise for cancer treatment. A new modular recombination strategy created novel CARs, with 28(NM)-BB(MC) CAR-T cells demonstrating enhanced anti-tumor activity and persistence.
Area of Science:
- Immunology
- Biotechnology
- Cancer Research
Background:
- Chimeric antigen receptor (CAR)-T cell therapy is a promising cancer treatment.
- CAR-T cell efficacy is often limited by T cell apoptosis and exhaustion.
- The intracellular signaling domain of CARs dictates T cell behavior.
Purpose of the Study:
- To engineer novel CARs with synthetic co-signaling modules using a modular recombination strategy.
- To characterize the signaling behaviors of these engineered CARs.
- To identify CAR designs that improve anti-tumor efficiency and T cell persistence.
Main Methods:
- Implemented a modular recombination strategy to construct a library of CARs.
- Incorporated synthetic co-signaling modules from the immunoglobulin-like superfamily (IgSF) and tumor necrosis factor receptor superfamily (TNFRSF).
- Quantitatively assessed CAR signaling using NFAT and NF-κB reporter assays.
Main Results:
- Successfully generated a library of CARs with diverse signaling profiles.
- Identified novel CAR constructs with distinct signaling behaviors.
- The 28(NM)-BB(MC) CAR-T cells demonstrated significantly improved cytotoxicity and enhanced T cell persistence.
Conclusions:
- The modular recombination approach provides a powerful tool for CAR-T cell engineering.
- This strategy enhances understanding of CAR signaling principles.
- Developed novel CARs with potential for improved cancer immunotherapy.
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