Immunization with mannosylated nanovaccines and inhibition of the immune-suppressing microenvironment sensitizes melanoma to immune checkpoint modulators

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Abstract

A low response rate, acquired resistance and severe side effects have limited the clinical outcomes of immune checkpoint therapy. Here, we show that combining cancer nanovaccines with an anti-PD-1 antibody (αPD-1) for immunosuppression blockade and an anti-OX40 antibody (αOX40) for effector T-cell stimulation, expansion and survival can potentiate the efficacy of melanoma therapy. Prophylactic and therapeutic combination regimens of dendritic cell-targeted mannosylated nanovaccines with αPD-1/αOX40 demonstrate a synergism that stimulates T-cell infiltration into tumours at early treatment stages. However, this treatment at the therapeutic regimen does not result in an enhanced inhibition of tumour growth compared to αPD-1/αOX40 alone and is accompanied by an increased infiltration of myeloid-derived suppressor cells in tumours. Combining the double therapy with ibrutinib, a myeloid-derived suppressor cell inhibitor, leads to a remarkable tumour remission and prolonged survival in melanoma-bearing mice. The synergy between the mannosylated nanovaccines, ibrutinib and αPD-1/αOX40 provides essential insights to devise alternative regimens to improve the efficacy of immune checkpoint modulators in solid tumours by regulating the endogenous immune response.

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Fig. 1: NP and man-NP are potential delivery systems for vaccination.
Fig. 2: NP and man-NP vaccines induce splenocyte activation and ex vivo cytotoxicity against melanoma cells.
Fig. 3: Prophylactic nanovaccines have a synergistic effect with PD-1 blockade and OX40 activation, which restricts melanoma growth and prolongs survival.
Fig. 4: Low CD8+:Treg ratio and high infiltration of MDSCs (CD11b+Gr-1+MDSC) compromise the therapeutic efficacy of the combination of mannosylated nanovaccines with αPD-1/αOX40.
Fig. 5: Trivalent combination of mannosylated nanovaccines with ibrutinib and αPD-1/αOX40 strongly restricts melanoma growth, which leads to long-term survival.
Fig. 6: Proposed model for the trivalent therapeutic strategy that combines mannosylated nanovaccines with ibrutinib and αPD-1/αOX40.

Data availability

The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information. Additional data and source files are available from the corresponding authors upon reasonable request.

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Acknowledgements

The MultiNano@MBM project was supported by The Israeli Ministry of Health and The Fundação para a Ciência e Tecnologia-Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES) under the framework of EuroNanoMed-II (ENMed/0051/2016 to H.F.F., R.S.-F. and S.J.). R.S.-F. thanks the European Research Council (ERC) Consolidator Grant Agreement no. [617445]-PolyDorm and ERC Advanced Grant Agreement no. [835227] - 3DBrainStrom, The Israel Science Foundation (Grant nos. 918/14 and 1969/18), The Melanoma Research Alliance (the Saban Family Foundation–MRA Team Science Award to R.S.-F. and N.E., and Established Investigator Award to R.S.-F.) and the Israel Cancer Research Fund (ICRF). J.C., A.I.M., C.P., E.Z. and L.I.F.M. are supported by the FCT-MCTES (Fellowships SFRH/BD/87150/2012, PD/BD/113959/2015, SFRH/BD/87591/2012, SFRH/BD/78480/2011 and SFRH/BPD/94111/2013, respectively). This project has received funding from European Structural & Investment Funds through the COMPETE Programme and from National Funds through FCT under the Programme grant SAICTPAC/0019/2015 (H.F.F.). We thank E. Haimov and B. Redko from the Blavatnik Center for Drug Discovery at the Tel Aviv University for their professional and technical assistance with the peptide synthesis.

Author information

J.C. and A.S. synthesized the nanovaccines and performed the in vitro and the animal studies; J.C. synthesized the man-PLGA polymers and carried out the physicochemical characterization of the nanovaccines; R.K. helped with the nanovaccine formulation; A.S., S.P. and R.K. performed the ELISPOT experiments; C.P. and A.I.M. analysed the flow cytometry experiments; E.Y. and S.P. performed the immunohistochemistry experiments; J.C. and A.I.M. carried out the tetramer assay; L.I.F.M. and E.Z. helped with the animal experiments; A.S.V. performed the AFM experiments; H.D. and N.E. contributed with the Ret cells; P.M.P.G. advised on the polymer synthesis; S.J. critically advised and contributed in interpreting the results. J.C., A.S., R.S.-F. and H.F.F. conceived and designed the experiments, analysed the data and wrote and revised the manuscript. R.S.-F. and H.F.F. were in charge of the overall direction and planning of this study, and all the authors commented on the manuscript.

Correspondence to Ronit Satchi-Fainaro or Helena F. Florindo.

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Peer review information: Nature Nanotechnology thanks Walter Storkus and other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Conniot, J., Scomparin, A., Peres, C. et al. Immunization with mannosylated nanovaccines and inhibition of the immune-suppressing microenvironment sensitizes melanoma to immune checkpoint modulators. Nat. Nanotechnol. 14, 891–901 (2019) doi:10.1038/s41565-019-0512-0

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