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Visualization of early events in mRNA vaccine delivery in non-human primates via PET–CT and near-infrared imaging

Nature Biomedical Engineering volume 3, pages 371–380 (2019)Cite this article

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Abstract

Visualization of the spatio–temporal trafficking of vaccines after their delivery would help evaluate the efficacy of candidate formulations and aid their rational design for preclinical and translational studies. Here, we show that a dual radionuclide–near-infrared probe allows for quantitative, longitudinal and non-invasive monitoring, via positron emission tomography–computed tomography and near-infrared imaging of cynomolgus macaques, of the trafficking dynamics to draining lymph nodes of a model messenger RNA vaccine labelled with the probe. After intramuscular administration of the vaccine to the monkeys, we observed the dynamics of the mRNA vaccine at the injection site and in the draining lymph nodes, performed cellular analyses of the involved tissues using flow cytometry and identified through immunofluorescence that professional antigen-presenting cells are the primary cells containing the injected mRNA and encoding the antigen. This approach may reveal spatio–temporal determinants of vaccine efficacy in preclinical and translational studies employing large mammals.

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Fig. 1: Experimental workflow and outline after i.m. injection with YF prME mRNA labelled with64Cu–DyLight 680.
Fig. 2: Approach to orthogonally label YF prME mRNA with dual radionuclide–near-IR probes.
Fig. 3: Representative PET–CT images of non-human primates 4 h after i.m. 64Cu vaccine delivery.
Fig. 4: PET–CT images of non-human primates 28 h after i.m. 64Cu vaccine delivery.
Fig. 5: Total SUV increases over 28 h in draining LNs.
Fig. 6: APCs are the primary cell type containing radiolabelled prME mRNA in muscle and LNs.
Fig. 7: YF prME mRNA and protein expression in LNs.

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Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information. The PET–CT-scanner acquisition settings are available on request from the corresponding author.

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Acknowledgements

The Petit Institute Core at Georgia Tech, particularly S. Durham and A. Shaw, who provided guidance and resources for flow cytometry and confocal microscopy, respectively. This work was supported by the Defense Advanced Research Projects Agency, Sanofi Pasteur and the RNArmorVax Consortium (to P.J.S). The views, opinions and/or findings expressed are those of the author(s) and should not be interpreted as representing the official views or policies of the US Department of Defense or the US Government.

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Author notes

  1. These authors contributed equally: Kevin E. Lindsay, Sushma M. Bhosle, Chiara Zurla.

Authors and Affiliations

  1. Wallace H. Coulter Department of Biomedical Engineering, Georgia Tech and Emory University, Atlanta, GA, USA

    Kevin E. Lindsay, Sushma M. Bhosle, Chiara Zurla, Jared Beyersdorf, Daryll Vanover & Philip J. Santangelo

  2. New Iberia Research Center, University of Louisiana at Lafayette, Lafayette, LA, USA

    Kenneth A. Rogers, Peng Xiao, Mariluz Araínga, Lisa M. Shirreff & Francois Villinger

  3. CRCINA, INSERM, University of Nantes, University of Angers, Nantes, France

    Bruno Pitard

  4. CureVac AG, Tübingen, Germany

    Patrick Baumhof

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Contributions

K.E.L. performed the probe labelling of mRNA, 64Cu-reporter labelling, necropsies, near-IR tissue extraction and processing, tissue imaging and quantification, PET–CT data analysis, and wrote the manuscript. S.M.B. performed in vitro transfections, probe labelling and optimization of mRNA labelling, nanoparticle optimization, tissue staining, imaging and quantification, and wrote the manuscript. C.Z. performed optimization of mRNA labelling with probes, HPLC of RNA, RT–PCR of innate gene expression, preparation of samples for flow cytometry, and paper revision and editing. J.B. analysed the flow cytometry data. K.A.R., P.X. and L.M.S. performed the flow cytometry. D.V. and M.A. performed PET imaging and necropsy. B.P. synthesized the CholK nanoparticles. P.B. synthesized the IVT YF prME mRNA. F.V. designed the experiments, and performed PET imaging, animal care and necropsies. P.J.S. designed the experiments, performed probe labelling of mRNA, PET imaging and edited the manuscript.

Corresponding author

Correspondence to Philip J. Santangelo.

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Competing interests

B.P. owns stock in In-Cell-Art, which commercializes lipidic aminoglycoside derivatives. P.B. is an employee at CureVac, which commercializes RNA-based vaccines.

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Supplementary information

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Supplementary Video

3D reconstruction of vaccine PET–CT signal at 28 hours post injection in AF093.

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Lindsay, K.E., Bhosle, S.M., Zurla, C. et al. Visualization of early events in mRNA vaccine delivery in non-human primates via PET–CT and near-infrared imaging. Nat Biomed Eng 3, 371–380 (2019). https://doi.org/10.1038/s41551-019-0378-3

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