|
Title
|
|
|
|
Design of an optimized Wilms' tumor 1 (WT1) mRNA construct for enhanced WT1 expression and improved immunogenicity in vitro and in vivo
| |
|
Author
|
|
|
|
| |
|
Abstract
|
|
|
|
| Tumor antigen-encoding mRNA for dendritic cell (DC)-based vaccination has gained increasing popularity in recent years. Within this context, two main strategies have entered the clinical trial stage: the use of mRNA for ex vivo antigen loading of DCs and the direct application of mRNA as a source of antigen for DCs in vivo. DCs transfected with mRNA-encoding Wilms' tumor 1 (WT1) protein have shown promising clinical results. Using a stepwise approach, we re-engineered a WT1 cDNA-carrying transcription vector to improve the translational characteristics and immunogenicity of the transcribed mRNA. Different modifications were performed: (i) the WT1 sequence was flanked by the lysosomal targeting sequence of dendritic cell lysosomal-associated membrane protein to enhance cytoplasmic expression; (ii) the nuclear localization sequence (NLS) of WT1 was deleted to promote shuttling from the nucleus to the cytoplasm; (iii) the WT1 DNA sequence was optimized in silico to improve translational efficiency; and (iv) this WT1 sequence was cloned into an optimized RNA transcription vector. DCs electroporated with this optimized mRNA showed an improved ability to stimulate WT1-specific T-cell immunity. Furthermore, in a murine model, we were able to show the safety, immunogenicity, and therapeutic activity of this optimized mRNA. This work is relevant for the future development of improved mRNA-based vaccine strategies K. |
| |
|
Language
|
|
|
|
English
| |
|
Source (journal)
|
|
|
|
Molecular therapy : Nucleic acids. - New York, NY, 2012, currens
| |
|
Publication
|
|
|
|
New York, NY
:
Nature Pub. Group
,
2013
| |
|
ISSN
|
|
|
|
2162-2531
| |
|
DOI
|
|
|
|
10.1038/MTNA.2013.54
| |
|
Volume/pages
|
|
|
|
2
(2013)
, 9 p.
| |
|
Article Reference
|
|
|
|
e134
| |
|
ISI
|
|
|
|
000332469000004
| |
|
Medium
|
|
|
|
E-only publicatie
| |
|
Full text (Publisher's DOI)
|
|
|
|
| |
|