Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo
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...
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doaj-a462f9b35af243399c03aa019a2d528a2020-11-25T00:44:47ZengElsevierMolecular Therapy: Nucleic Acids2162-25312013-01-012C10.1038/mtna.2013.54Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In VivoDaphné Benteyn0Sébastien Anguille1Sandra Van Lint2Carlo Heirman3An MT Van Nuffel4Jurgen Corthals5Sebastian Ochsenreither6Wim Waelput7Katrien Van Beneden8Karine Breckpot9Viggo Van Tendeloo10Kris Thielemans11Aude Bonehill12Laboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumCenter for Cell Therapy & Regenerative Medicine, Antwerp University Hospital, Antwerp, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumFred Hutchinson Cancer Research Center, Seattle, WashingtonDepartment of Pathology, Universitair Ziekenhuis Brussel, Brussels, BelgiumLiver Cell Biology Laboratory, Department of Human Anatomy, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Experimental Hematology, Tumor Immunology Group (TIGR), Vaccine & Infectious Disease Institute, University of Antwerp, Antwerp, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Molecular & Cellular Therapy, Vrije Universiteit Brussel, Brussels, BelgiumTumor 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.http://www.sciencedirect.com/science/article/pii/S2162253116301901 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Daphné Benteyn Sébastien Anguille Sandra Van Lint Carlo Heirman An MT Van Nuffel Jurgen Corthals Sebastian Ochsenreither Wim Waelput Katrien Van Beneden Karine Breckpot Viggo Van Tendeloo Kris Thielemans Aude Bonehill |
spellingShingle |
Daphné Benteyn Sébastien Anguille Sandra Van Lint Carlo Heirman An MT Van Nuffel Jurgen Corthals Sebastian Ochsenreither Wim Waelput Katrien Van Beneden Karine Breckpot Viggo Van Tendeloo Kris Thielemans Aude Bonehill Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo Molecular Therapy: Nucleic Acids |
author_facet |
Daphné Benteyn Sébastien Anguille Sandra Van Lint Carlo Heirman An MT Van Nuffel Jurgen Corthals Sebastian Ochsenreither Wim Waelput Katrien Van Beneden Karine Breckpot Viggo Van Tendeloo Kris Thielemans Aude Bonehill |
author_sort |
Daphné Benteyn |
title |
Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo |
title_short |
Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo |
title_full |
Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo |
title_fullStr |
Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo |
title_full_unstemmed |
Design of an Optimized Wilms’ Tumor 1 (WT1) mRNA Construct for Enhanced WT1 Expression and Improved Immunogenicity In Vitro and In Vivo |
title_sort |
design of an optimized wilms’ tumor 1 (wt1) mrna construct for enhanced wt1 expression and improved immunogenicity in vitro and in vivo |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2013-01-01 |
description |
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. |
url |
http://www.sciencedirect.com/science/article/pii/S2162253116301901 |
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