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Synthetic Messenger RNA and Cell Metabolism Modulation pp 247–274Cite as

Mannosylated and Histidylated LPR Technology for Vaccination with Tumor Antigen mRNA

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 969))

Abstract

mRNA-based vaccines are currently being developed for treating various diseases including cancers. For this purpose, synthetic or in vitro transcribed (IVT) mRNA encoding tumor antigen offers several advantages over plasmid DNA encoding tumor antigen including better delivery and security. In this chapter, we report the preparation of mannosylated mRNA nanoparticles termed mannosylated lipopolyplexes or Man-LPR loaded with mRNA encoding a melanoma antigen. This formulation enhances the transfection of dendritic cells (DCs) in vivo and the anti-B16F10 melanoma vaccination in mice. The mRNA is formulated with histidylated liposomes and a histidylated polymer. Those pH-sensitive vectors promote membrane destabilization in endosomes upon the protonation of their histidine groups, allowing nucleic acid delivery in the cytosol. To favor DCs targeting via the mannose receptor, a mannose lipid is incorporated in the liposomes. Here, we provide protocols for the preparation of mannosylated liposomes, the synthesis of mRNA, mice immunization based on systemic injection, measurement of the cellular immune response and determination of the number of transfected splenic DC.

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Acknowledgments

We are indebted to Ligue Nationale contre le Cancer, Ligue contre le Cancer (Région Centre), and Canceropole Grand Ouest for the funding of our research project. We gratefully acknowledge the members of our team, especially Cristine Gonçalves and our former PhD. and Master Students (Michael Mockey, Erika Bourseau and Federico Perche) which allowed the realization of these projects. We also thank Paul-Alain Jaffres, Mathieu Mével and Mathieu Berchel (from Brest), and Thierry Benvegnu and Marie-Laure Miramon (from Rennes).

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Authors and Affiliations

  1. Centre de Biophysique Moléculaire, CNRS UPR4301, Orléans, France

    Chantal Pichon & Patrick Midoux

  2. Université d’Orléans, Orléans, France

    Patrick Midoux

Authors
  1. Chantal Pichon
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  2. Patrick Midoux
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Correspondence to Patrick Midoux .

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Editors and Affiliations

  1. The Anlyan Center, Department of Genetics, Yale School of Medicine, Cedar Street 300, New Haven, 06520-8005, Connecticut, USA

    Peter M. Rabinovich

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Pichon, C., Midoux, P. (2013). Mannosylated and Histidylated LPR Technology for Vaccination with Tumor Antigen mRNA. In: Rabinovich, P. (eds) Synthetic Messenger RNA and Cell Metabolism Modulation. Methods in Molecular Biology, vol 969. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-260-5_16

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  • DOI: https://doi.org/10.1007/978-1-62703-260-5_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-259-9

  • Online ISBN: 978-1-62703-260-5

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