Bibliographic Details
| Title: |
Efficient and Non-genotoxic RNA-Based Engineering of Human T Cells Using Tumor-Specific T Cell Receptors With Minimal TCR Mispairing |
| Authors: |
Diana Campillo-Davo, Fumihiro Fujiki, Johan M. J. Van den Bergh, Hans De Reu, Evelien L. J. M. Smits, Herman Goossens, Haruo Sugiyama, Eva Lion, Zwi N. Berneman, Viggo Van Tendeloo |
| Source: |
Frontiers in Immunology, Vol 9 (2018) |
| Publisher Information: |
Frontiers Media S.A., 2018. |
| Publication Year: |
2018 |
| Collection: |
LCC:Immunologic diseases. Allergy |
| Subject Terms: |
TCR-gene transfer, electroporation, adoptive cell therapy (ACT), DsiRNA, RNA transfection, Immunologic diseases. Allergy, RC581-607 |
| More Details: |
Genetic engineering of T cells with tumor specific T-cell receptors (TCR) is a promising strategy to redirect their specificity against cancer cells in adoptive T cell therapy protocols. Most studies are exploiting integrating retro- or lentiviral vectors to permanently introduce the therapeutic TCR, which can pose serious safety issues when treatment-related toxicities would occur. Therefore, we developed a versatile, non-genotoxic transfection method for human unstimulated CD8+ T cells. We describe an optimized double sequential electroporation platform whereby Dicer-substrate small interfering RNAs (DsiRNA) are first introduced to suppress endogenous TCR α and β expression, followed by electroporation with DsiRNA-resistant tumor-specific TCR mRNA. We demonstrate that double sequential electroporation of human primary unstimulated T cells with DsiRNA and TCR mRNA leads to unprecedented levels of transgene TCR expression due to a strongly reduced degree of TCR mispairing. Importantly, superior transgenic TCR expression boosts epitope-specific CD8+ T cell activation and killing activity. Altogether, DsiRNA and TCR mRNA double sequential electroporation is a rapid, non-integrating and highly efficient approach with an enhanced biosafety profile to engineer T cells with antigen-specific TCRs for use in early phase clinical trials. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
1664-3224 |
| Relation: |
https://www.frontiersin.org/article/10.3389/fimmu.2018.02503/full; https://doaj.org/toc/1664-3224 |
| DOI: |
10.3389/fimmu.2018.02503 |
| Access URL: |
https://doaj.org/article/c139e17698ca4d20b686af4b4ae43c01 |
| Accession Number: |
edsdoj.139e17698ca4d20b686af4b4ae43c01 |
| Database: |
Directory of Open Access Journals |
