Abstract
Dendritic cells (DCs) are the orchestrators of the immune system and are frequently used in clinical trials in order to boost the immune system in cancer patients. Among several available techniques for DC modification, mRNA electroporation is an interesting technique due to the favorable characteristics of mRNA. Antigen expression level and duration can be increased by multiple optimizations of an antigen-encoding mRNA template. Here, we describe different molecular modifications to a WT1-encoding mRNA construct in order to increase antigen expression and the subsequent introduction of mRNA into DCs.
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References
Benteyn D, Heirman C, Bonehill A et al (2015) mRNA-pulsed dendritic cell vaccines. Expert Rev Vaccines 14:161–176
Cheever MA, Allison JP, Ferris AS et al (2009) The prioritization of cancer antigens: a national cancer institute pilot project for the acceleration of translational research. Clin Cancer Res 15:5323–5337
Bonehill A, Heirman C, Tuyaerts S et al (2004) Messenger RNA-electroporated dendritic cells presenting MAGE-A3 simultaneously in HLA class I and class II molecules. J Immunol 172:6649–6657
Benteyn D, Anguille S, Van Lint S et al (2013) Design of an optimized Wilms’ tumor 1 (WT1) mRNA construct for enhanced WT1 expression and improved immunogenicity in vitro and in vivo. Mol Ther Nucleic Acids 2:e134
De Keersmaecker B, Heirman C, Allard S et al (2010) Lumenal part of the DC-LAMP protein is not required for induction of antigen-specific T cell responses by means of antigen-DC-LAMP messenger RNA-electroporated dendritic cells. Hum Gene Ther 21:479–485
Kreiter S, Selmi A, Diken M et al (2008) Increased antigen presentation efficiency by coupling antigens to MHC class I trafficking signals. J Immunol 180:309–318
Niksic M, Slight J, Sanford JR et al (2004) The Wilms’ tumour protein (WT1) shuttles between nucleus and cytoplasm and is present in functional polysomes. Hum Mol Genet 13:463–471
Kuhn AN, Diken M, Kreiter S et al (2012) Determinants of intracellular RNA pharmacokinetics: Implications for RNA-based immunotherapeutics. RNA Biol 8:35–43
Pascolo S (2008) Vaccination with messenger RNA (mRNA). Handb Exp Pharmacol 183:221–235
Fath S, Bauer AP, Liss M et al (2011) Multiparameter RNA and codon optimization: a standardized tool to assess and enhance autologous mammalian gene expression. PLoS One 6:e17596
Turksma AW, Bontkes HJ, Ruizendaal JJ et al (2013) Increased cytotoxic capacity of tumor antigen specific human T cells after in vitro stimulation with IL21 producing dendritic cells. Hum Immunol 74:506–513
Holtkamp S, Kreiter S, Selmi A et al (2006) Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. Blood 108:4009–4017
Van Driessche A, Van de Velde ALR, Nijs G et al (2013) Clinical-grade manufacturing of autologous mature mRNA-electroporated dendritic cells and safety testing in acute myeloid leukemia patients in a phase I dose-escalation clinical trial. Cytotherapy 11:653–668
Melhem NM, Gleason SM, Liu XD et al (2014) High-level antigen expression and sustained antigen presentation in dendritic cells nucleofected with wild-type viral mRNA but not DNA. Clin Vaccine Immunol 15:1337–1344
De Haes W, Rejman J, Pollard C et al (2013) Lipoplexes carrying mRNA encoding Gag protein modulate dendritic cells to stimulate HIV-specific immune responses. Nanomedicine 8:77–87
De Temmerman M-L, Dewitte H, Vandenbroucke RE et al (2011) mRNA-Lipoplex loaded microbubble contrast agents for ultrasound-assisted transfection of dendritic cells. Biomaterials 32:9128–9135
Acknowledgments
This work was supported by grants from the Interuniversity Attraction Poles Program—Belgian State—Belgian Science Policy, the National Cancer Plan of the Federal Ministry of Hearlth, the Stichting tegen Kanker, the Vlaamse Liga tegen Kanker, an Integrated Project and a Network of Excellence sponsored by the EU FP-6, an IWT-TBM program, the Fonds voor Wetenschappelijk Onderzoek Vlaanderen (FWO-Vlaanderen), and the Willy Gepts Wetenschappelijk Fonds of the UZ Brussel.
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Benteyn, D., Heirman, C., Thielemans, K., Bonehill, A. (2016). Engineering WT1-Encoding mRNA to Increase Translational Efficiency in Dendritic Cells. In: Rhoads, R. (eds) Synthetic mRNA. Methods in Molecular Biology, vol 1428. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3625-0_7
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DOI: https://doi.org/10.1007/978-1-4939-3625-0_7
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3623-6
Online ISBN: 978-1-4939-3625-0
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