|
Title
|
|
|
|
Efficient and non-genotoxic RNA-based engineering of human T cells using tumor-specific T cell receptors with minimal TCR mispairing
| |
|
Author
|
|
|
|
| |
|
Abstract
|
|
|
|
| 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 sequential double 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 sequential double 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. |
| |
|
Language
|
|
|
|
English
| |
|
Source (journal)
|
|
|
|
Frontiers in immunology. - Place of publication unknown
Frontiers in immunology. - Place of publication unknown
| |
|
Publication
|
|
|
|
Place of publication unknown
:
2018
| |
|
ISSN
|
|
|
|
1664-3224
| |
|
Volume/pages
|
|
|
|
9
(2018)
, 14 p.
| |
|
Article Reference
|
|
|
|
2503
| |
|
ISI
|
|
|
|
000449420000001
| |
|
Pubmed ID
|
|
|
|
30464762
| |
|
Medium
|
|
|
|
E-only publicatie
| |
|
Full text (Publisher's DOI)
|
|
|
|
| |
|
Full text (open access)
|
|
|
|
| |
|