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Updated: Aug 9, 2025
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A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy
Published on: February 21, 2025
702
Engineered exosome-mediated messenger RNA and single-chain variable fragment delivery for human chimeric antigen
Ke Si1, Zhu Dai1, Zhanping Li1
1State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China.
Cytotherapy
|February 24, 2023
Summary
Researchers developed an exosome delivery platform to produce chimeric antigen receptor (CAR) T cells using messenger RNA (mRNA). This novel method bypasses viral transduction and electroporation toxicity, enabling efficient CAR T cell generation for cancer therapy.
Area of Science:
- Biotechnology
- Immunotherapy
- Cellular Engineering
Background:
- Current chimeric antigen receptor (CAR) T cell therapies often rely on viral transduction, which carries risks of adverse effects.
- Messenger RNA (mRNA)-based CAR T cell production offers an alternative but faces challenges with current delivery methods like electroporation, which can be toxic.
Purpose of the Study:
- To engineer a novel exosome-based delivery platform for mRNA encoding CARs and T cell activators.
- To evaluate the efficiency and efficacy of this exosome platform for CAR T cell generation.
Main Methods:
- Designed an exosome delivery system incorporating the bacteriophage MS2 system and lysosome-associated membrane protein 2 isoform B.
- Loaded exosomes with mRNA encoding CARs and anti-CD3/CD28 single-chain variable fragments (scFvs).
- Assessed mRNA loading, delivery, protein expression, and T cell activation.
Main Results:
- The engineered exosomes successfully loaded and delivered mRNA into target cells, leading to protein expression.
- Exosomes displaying anti-CD3/CD28 scFvs effectively activated primary T cells, comparable to commercial magnetic beads.
- Demonstrated ex vivo production of CAR T cells with cancer cell-killing capabilities.
Conclusions:
- Engineered exosomes provide an effective platform for delivering CAR mRNA and activating scFvs for CAR T cell production.
- This exosome-based approach offers a potentially safer and more efficient alternative to viral transduction and electroporation.
- The platform holds promise for both ex vivo CAR T cell manufacturing and in vivo CAR T cell generation strategies.
