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Cancer vaccines from cryogenically silicified tumour cells functionalized with pathogen-associated molecular patterns

Nature Biomedical Engineering volume 6, pages 19–31 (2022)Cite this article

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Abstract

The production of personalized cancer vaccines made from autologous tumour cells could benefit from mechanisms that enhance immunogenicity. Here we show that cancer vaccines can be made via the cryogenic silicification of tumour cells, which preserves tumour antigens within nanoscopic layers of silica, followed by the decoration of the silicified surface with pathogen-associated molecular patterns. These pathogen-mimicking cells activate dendritic cells and enhance the internalization, processing and presentation of tumour antigens to T cells. In syngeneic mice with high-grade ovarian cancer, a cell-line-based silicified cancer vaccine supported the polarization of CD4+ T cells towards the T-helper-1 phenotype in the tumour microenvironment, and induced tumour-antigen-specific T-cell immunity, resulting in complete tumour eradication and in long-term animal survival. In the setting of established disease and a suppressive tumour microenvironment, the vaccine synergized with cisplatin. Silicified and surface-modified cells from tumour samples are amenable to dehydration and room-temperature storage without loss of efficacy and may be conducive to making individualized cancer vaccines across tumour types.

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Fig. 1: Characterization of Si cancer cells.
Fig. 2: Surface functionalization enhances DC uptake and activation in vitro.
Fig. 3: Treatment with Si cells induces a protective immune response in vivo.
Fig. 4: Therapeutic vaccination clears pre-existing tumours.
Fig. 5: Therapeutic benefit of vaccination is associated with significant changes in tumour-associated lymphocytes and cytokines.
Fig. 6: Combination cisplatin and vaccine therapy clears established peritoneal tumours and enhances survival in mice.
Fig. 7: Effective vaccine can be developed from ascites.

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Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. The raw and analysed datasets generated during the study are too large to be publicly shared, but they are available for research purposes from the corresponding author on reasonable request. Source data for the figures are provided with this paper.

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Acknowledgements

We thank the University of New Mexico for the assistance and use of the Comprehensive Cancer Center Animal Models, Fluorescence Microscopy, Flow Cytometry and Histology Shared Resources, supported by NIH grant NCI P30 CA118100 (PI, Willman C.). This work was also supported by AIM center cores funded by NIH grant P20GM121176. We thank F. Schultz, I. Lagutina and M. Steinkamp for technical assistance.

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Author notes

  1. These authors contributed equally: Jimin Guo, Henning De May.

Authors and Affiliations

  1. Department of Internal Medicine, University of New Mexico Health Science Center, Albuquerque, NM, USA

    Jimin Guo, Stefan Franco, Donna F. Kusewitt & Rita E. Serda

  2. Department of Obstetrics & Gynecology, University of New Mexico Health Science Center, Albuquerque, NM, USA

    Henning De May & Sarah F. Adams

  3. Department of Chemical and Biological Engineering, University of New Mexico, Albuquerque, NM, USA

    Achraf Noureddine, Lien Tang & C. J. Brinker

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Contributions

R.E.S. and S.F.A. designed and co-directed the study. S.F., A.N. and L.T. conducted research. R.E.S., S.F.A., J.G., H.D.M. and D.F.K. contributed to the experimental design and conducted research. All authors reviewed the manuscript.

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Correspondence to Rita E. Serda.

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Competing interests

R.E.S., S.F.A., J.G. and C.J.B. are inventors on patent applications (US patent application no. 20200276286) based on technology presented in this manuscript.

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Peer review information Nature Biomedical Engineering thanks Sidi Bencherif, Lélia Delamarre and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Therapeutic efficacy of single vs dual TLR ligand vaccines.

a) Timeline for BR5-Akt-Luc2 cancer cell and vaccine administration. b) Chitosan (CHIT) or polyethylimine (PEI) vaccine formulations containing MPL; CpG; or MPL plus CpG. c) IVIS bioluminescence images of FVB mice over time. d) Average tumour burden (photons/second; p/s) of each group shown in ‘c’ on Day 19 (Unpaired, two-tailed, parametric t-tests, s.d. error bars). e) Kaplan-Meier survival curves for mice treated with Si-PEI cells coated with CpG, or CpG and MPL. f = free, and b = bound TLR ligand; (n = 4/group, Log-rank Mantel-Cox: p < 0.0001 with symbols to the right on the survival curve representing comparisons with no treatment (no Tx) controls and the symbol on the purple line comparing the CpG vs CpG/MPL (f + b) groups. * p < 0.05, **p < 0.01, p < 0.001.

Extended Data Fig. 2 Bone marrow-derived dendritic cell (BMDC) activation and internalization of silicified (Si) cells.

a) 2D and 3D confocal images show five internalized Si-LPS-ID8ova cells (visualized via internalized DyLight 488-labelled nanoparticles) in a single BMDC. b) Flow cytometry analysis of SIINFEKL MHC-I (H-2Kb) presentation and surface expression of CD80 in BMDC incubated with Si cells presenting null, PEI, or PEI plus LPS or MPL for 24 hours (n = 3/group; ***p < 0.001). c) Cell Trace Far Red-labelled vaccine cells were predominately located in filtering and lymphatic organs (LN-M: mesenteric; LNs: Inguinal and axillary) 24 and 48 h post IP injection in tumour-bearing mice (n = 2-3 biological replicates). d) Fluorescent vaccine cells (red; Cy3) were located in peritoneal lymph nodes and the omentum, predominately in DC-rich areas, 24-48 hours post IP injection in naïve mice (green, right; CD11c FITC; blue nuclei, DAPI). Unpaired, two-tailed, parametric t-tests, SD error bars.

Extended Data Fig. 3 Therapeutic vaccination in the tumour microenvironment enables systemic immunity.

a) Experimental design and timeline (n = 3). b) IVIS bioluminescent images of BR5-Akt-Luc2 tumour growth. c) Graph of tumour burden (photons/sec) across time by treatment group (no treatment (no Tx) and tumour location. Intraperitoneal (IP) vaccination of mice bearing both subcutaneous (SC) and IP tumours cleared established tumours in both locations while mice with only SC tumours continued to display tumour growth (n = 3/group, Holm-Sidak multiple comparison; individual animal curves in SI). **p < 0.01.

Extended Data Fig. 4 Vaccination activates T cells in the tumour microenvironment.

To test the effect of treatment number on IP immune responses, IP cells were collected and analysed for activation and functional status using flow cytometry on Day 19 following tumour challenge in mice vaccinated 1x, 2x, 3x, 4x or those receiving no treatment (PBS; no Tx) on days 4, 6,11 and 17. a) Proportion of IP CD4 and CD8 cells with effector memory phenotypes (CD44+ CD62Llow). b) Representative dot plot for effector memory CD4+ IP cells (CD44 vs CD62L. c) Representative plot for IFNγ expression by IP CD4+ cells from with no vaccination vs 2x vaccination. d) Proportion of IP CD4+ cells expressing the regulatory markers CTLA4 and FoxP3. e) Proportion of CD4+ and CD8+ IP cells expressing PD1 and Tim3. n = 3 biological replicates; unpaired, two-tailed, parametric t-tests, SD error bars, *p < 0.05, **p < 0.01, ***p < 0.001.

Extended Data Fig. 5 CBC and blood metabolite analysis supports vaccine safety.

Tumour naïve (a,c) or cancer challenged (b,d) FVB mice (n = 3 biological replicates; unpaired, two-tailed, parametric t-tests) were vaccinated 1-4x. Graphs show cell counts by population and blood metabolites obtained using the Abaxis VetScan System and piccolo metabolite discs. *p < 0.05, ***p < 0.001.

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Guo, J., De May, H., Franco, S. et al. Cancer vaccines from cryogenically silicified tumour cells functionalized with pathogen-associated molecular patterns. Nat Biomed Eng 6, 19–31 (2022). https://doi.org/10.1038/s41551-021-00795-w

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