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Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels

Abstract

Cancer recurrence can arise owing to rare circulating cancer stem cells (CSCs) that are resistant to chemotherapies and radiotherapies. Here, we show that a double-network hydrogel can rapidly reprogramme differentiated cancer cells into CSCs. Spheroids expressing elevated levels of the stemness genes Sox2, Oct3/4 and Nanog formed within 24 h of seeding the gel with cells from any of six human cancer cell lines or with brain cancer cells resected from patients with glioblastoma. Human brain cancer cells cultured on the double-network hydrogel and intracranially injected in immunodeficient mice led to higher tumorigenicity than brain cancer cells cultured on single-network gels. We also show that the double-network gel induced the phosphorylation of tyrosine kinases, that gel-induced CSCs from primary brain cancer cells were eradicated by an inhibitor of the platelet-derived growth factor receptor, and that calcium channel receptors and the protein osteopontin were essential for the regulation of gel-mediated induction of stemness in brain cancer cells.

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Fig. 1: DN gels rapidly induced CSCs of six human cancer cell lines.
Fig. 2: DN gels rapidly induced CSCs of human brain cancer cell lines and primary culture cells.
Fig. 3: Analysis of hydrogels for induction of CSCs and comparison of expression profiles of CSCs induced by DN gel and neurosphere culture conditions.
Fig. 4: Tumorigenic potential of DN gel-induced sphere-forming brain cancer cells in vivo.
Fig. 5: Analysis of DN gel-activated intracellular signalling pathways.
Fig. 6: Requirement for OPN for induction of brain CSCs by DN gels.
Fig. 7: DN gels induced PDGFR-expressing CSCs, which were eradicated by PDGFR inhibitor.
Fig. 8: Molecular mechanisms of the hydrogel-activated reprogramming phenomenon.

Data availability

The main data supporting the results of this study are available within the paper and its Supplementary Information. All raw data used to make the graphs are available from Figshare with the identifier https://doi.org/10.6084/m9.figshare.13297913.v1.

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Acknowledgements

We thank K. Hida (Hokkaido University, Japan) for the pCSII-CMV-tdTomato-Luc2 plasmid, S. Kon (Fukuyama University, Japan) for the anti-OPN antibody (clone 35B6), M. Sudol (National University of Singapore) for p2×FLAG-CMV2-YAP1, K. Tabuchi (Saga University, Japan) for the GBM cell line KMG4, A. Kawai (National Cancer Center, Japan) for the synovial sarcoma cell line SYO-1, A. Hirota (Hokkaido University, Japan) for the human induced pluripotent stem cells and Y. Hane (Hokkaido University) for gel synthesis. We also thank the medical students R. Nabeshima, K. Aoyama, T. Kurai and T. Ishizuka (Hokkaido University) for useful discussions. This work is supported by the Global Station for Soft Matter (a project of the Global Institution for Collaborative Research and Education at Hokkaido University) and, in part, by grants from MEXT (19H01171 to S.T., and 15K15106 and 18K07059 to M.T.), and AMED (20cm0106571h0001 and 21cm0106571h0002 to S.T.). The Institute for Chemical Reaction Design and Discovery (ICReDD) was established by the World Premier International Research Center Initiative (WPI), MEXT, Japan. This study used IVIS with support from the Global Center for Biomedical Science and Engineering, Faculty of Medicine, Hokkaido University.

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Contributions

J.S. performed all of the in vitro experiments with supervision from M.T. and S.S. L.W. performed the animal experiments. S.A. and Y.O. performed the microarray analysis. H.S. performed the calcium ion-channel analysis. T.K. and M.F. generated the hydrogels. K.Y. and J.P.G. organized the projects related to hydrogels at Hokkaido University. S. Kohsaka, S.Kojima, T.U. and H.M. performed the single-cell whole-transcriptome analysis. K.K. performed the immunofluorescence analysis. S.T. designed the entire study and wrote the manuscript.

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Correspondence to Shinya Tanaka.

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Supplementary information

Supplementary Information

Supplementary methods, figures, tables and video captions.

Reporting Summary

Supplementary Video 1

Rapid induction of spheroid formation on a DN gel.

Supplementary Video 2

SOX2-expressing cells in a spheroid formed on a DN gel.

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Suzuka, J., Tsuda, M., Wang, L. et al. Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels. Nat Biomed Eng (2021). https://doi.org/10.1038/s41551-021-00692-2

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