In situ vaccination with cowpea mosaic virus nanoparticles suppresses metastatic cancer

Abstract

Nanotechnology has tremendous potential to contribute to cancer immunotherapy. The ‘in situ vaccination’ immunotherapy strategy directly manipulates identified tumours to overcome local tumour-mediated immunosuppression and subsequently stimulates systemic antitumour immunity to treat metastases. We show that inhalation of self-assembling virus-like nanoparticles from cowpea mosaic virus (CPMV) reduces established B16F10 lung melanoma and simultaneously generates potent systemic antitumour immunity against poorly immunogenic B16F10 in the skin. Full efficacy required Il-12, Ifn-γ, adaptive immunity and neutrophils. Inhaled CPMV nanoparticles were rapidly taken up by and activated neutrophils in the tumour microenvironment as an important part of the antitumour immune response. CPMV also exhibited clear treatment efficacy and systemic antitumour immunity in ovarian, colon, and breast tumour models in multiple anatomic locations. CPMV nanoparticles are stable, nontoxic, modifiable with drugs and antigens, and their nanomanufacture is highly scalable. These properties, combined with their inherent immunogenicity and demonstrated efficacy against a poorly immunogenic tumour, make CPMV an attractive and novel immunotherapy against metastatic cancer.

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Figure 1: eCPMV nanoparticles are inherently immuonogenic.
Figure 2: eCPMV inhalation induces dramatic changes in lung immune cell composition and cytokine/chemokine milieu in mice bearing B16F10 lung tumours.
Figure 3: eCPMV inhalation reduces formation of B16F10 metastatic-like lung tumours.
Figure 4: eCPMV treatment efficacy in B16F10 lung model is immune-mediated.
Figure 5: eCPMV immunotherapy is successful in metastatic breast, colon, and ovarian carcinoma models.
Figure 6: eCPMV induces systemic, durable antitumour immunity.

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Acknowledgements

George P. Lomonossoff (John Innes Centre, UK) is thanked for providing the pEAQexpress-VP60-24K plasmid. We thank the following for their assistance: Zachary Parker; the lab of David Mullins; the lab of Brent Berwin; the Immune Monitoring Lab and DartLab at the Geisel School of Medicine at Dartmouth, particularly Gary Ward and John DeLong; the Dartmouth Transgenic and Genetic shared resource; the Dartmouth Hitchcock Medical Center pathology department; the Irradiation, Pre-clinical Imaging, and Microscopy (IPIM) and Genomics/Molecular Biology (GMB) shared resources at the Norris Cotton Cancer Center. Shared resources at Dartmouth are made possible through generous Centers of Biomedical Research Excellence support. Work was supported by Dartmouth Immunobiology of Myeloid and Lymphoid Cells National Institutes of Health Training Grant 5T32AI007363–22 (P.H.L.), Case Western Reserve University Cardiovascular Research National Institutes of Health Training Grant T32 HL105338 (A.M.W.), National Science Foundation CMMI 1333651 (N.F.S.), Dartmouth Center of Nanotechnology Excellence NIH 1 U54 CA151662 (S.F.), Center for Molecular, Cellular, and Translational Immunological Research NIGMS 1P30RR032136–01 (S.F.), and Norris Cotton Cancer Center P30 CA023108-27 (S.F.).

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P.H.L., N.F.S., and S.F. conceived and designed the experiments, and wrote the manuscript. P.H.L., A.M.W., P.R., M.R.S., and J.F. performed the experiments. M.R.S. was responsible for Supplementary Fig. 4. P.R. was responsible for Supplementary Fig. 6. A.M.W. was responsible for Supplementary Fig. 8. P.H.L. performed all other experiments and analysed the data. J.F. assisted with in vivo work. All authors commented on the manuscript.

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Correspondence to N. F. Steinmetz or S. Fiering.

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The authors have a patent pending for the immunotherapeutic use of the eCPMV nanoparticle.

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Lizotte, P., Wen, A., Sheen, M. et al. In situ vaccination with cowpea mosaic virus nanoparticles suppresses metastatic cancer. Nature Nanotech 11, 295–303 (2016). https://doi.org/10.1038/nnano.2015.292

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