Engineered exosome-mediated messenger RNA and single-chain variable fragment delivery for human chimeric antigen receptor T-cell engineering
Abstract
Background aims Most current chimeric antigen receptor (CAR) T cells are generated by viral transduction, which induces persistent expression of CARs and may cause serious undesirable effects. Messenger RNA (mRNA)-based approaches in manufacturing CAR T cells are being developed to overcome these challenges. However, the most common method of delivering mRNA to T cells is electroporation, which can be toxic to cells.Methods The authors designed and engineered an exosome delivery platform using the bacteriophage MS2 system in combination with the highly expressed protein lysosome-associated membrane protein 2 isoform B on exosomes.Results The authors' delivery platform achieved specific loading and delivery of mRNA into target cells and achieved expression of specific proteins, and anti-CD3/CD28 single-chain variable fragments (scFvs) expressed outside the exosomal membrane effectively activated primary T cells in a similar way to commercial magnetic beads.Conclusions The delivery of CAR mRNA and anti-CD3/CD28 scFvs via designed exosomes can be used for ex vivo production of CAR T cells with cancer cell killing capacity. The authors' results indicate the potential applications of the engineered exosome delivery platform for direct conversion of primary T cells to CAR T cells while providing a novel strategy for producing CAR T cells in vivo.
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These findings demonstrate the prospective application of engineered exosome delivery platforms in converting primary T cells into CAR T cells directly, furnishing a new approach for in vivo CAR T cell production.
“These findings demonstrate the prospective application of engineered exosome delivery platforms in converting primary T cells into CAR T cells directly, furnishing a new approach for in vivo CAR T cell production [106].”
From Exosome for mRNA delivery: Strategies and therapeutic applications (2024) · cites this paper as [106]
Engineered exosomes deliver CAR mRNA and anti-CD3/CD28 scFv for the ex vivo production of CAR T cells with tumor-killing ability.
“Engineered exosomes deliver chimeric antigen receptor (CAR) mRNA and anti-CD3/CD28 single-chain variable fragments (scFv) for the ex vivo production of CAR T cells with tumor-killing ability.”
From Exosome for mRNA delivery: Strategies and therapeutic applications (2024) · cites this paper as [106]
Phage MS2 system and LAMP-2B modified exosomes demonstrated ability to specifically load and deliver anti-CD3/CD28 scFv and CAR mRNA.
“This system demonstrated the ability to specifically load and deliver anti-CD3/CD28 scFv and CAR mRNA.”
From Exosome for mRNA delivery: Strategies and therapeutic applications (2024) · cites this paper as [106]
This engineered exosome delivery system has great potential for converting primary T cells into CAR T cells and provides a new approach for in vivo induction of CAR T cells.
“The results suggested that this engineered exosome delivery system has great potential for converting primary T cells into CAR T cells and provides a new approach for the in vivo induction of CAR T cells [106].”
From Exosome for mRNA delivery: Strategies and therapeutic applications (2024) · cites this paper as [106]
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- Authors
- Si, K., Dai, Z., Li, Z., Ye, Z., Ding, B., Feng, S., Sun, B., Shen, Y., & Xiao, Z.
- Affiliations
- State Key Laboratory of Digital Medical Engineering (CN)
- Zhongda Hospital Southeast University (CN)
- Corresponding author
- Shen, Yang
- Xiao, Zhongdang
- Venue
- Cytotherapy
- Publisher
- Elsevier BV
- Published
- 2023-06
- Volume
- 25
- Issue
- 6
- Pages
- 615-624
- ISSN
- 1465-3249 (ISSN-L)
- OpenAlex
- W4321601872
- Cited by
- 60
- References
- 53
- Citations by year
- 2026: 20 · 2025: 22 · 2024: 10 · 2023: 8
- FWCI
- 7.32
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Cytotherapy (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Fundamental Research Funds for the Central Universities (2242018K3DN05)
- Jiangsu Commission of Health (ZDA2020012)
- National Natural Science Foundation of China (81671807, 82072078)
- Southeast University
- Jiangsu Provincial Key Research and Development Program (BE2020777, SBE2020741118)
- Topics
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- engineered exosomes, chimeric antigen receptor T cells, bacteriophage MS2, Lamp2b, scFv, T cell activation, tumor cell killing
- MeSH terms
- Receptors, Chimeric Antigen, Humans, Receptors, Antigen, T-Cell, T-Lymphocytes, Immunotherapy, Adoptive, CD28 Antigens, Cell Line, Tumor, Exosomes, Single-Chain Antibodies, Cell Engineering
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