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A three-dimensional engineered tumour for spatial snapshot analysis of cell metabolism and phenotype in hypoxic gradients

Nature Materials volume 15, pages 227–234 (2016)Cite this article

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An Erratum to this article was published on 22 January 2016

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Abstract

The profound metabolic reprogramming that occurs in cancer cells has been investigated primarily in two-dimensional cell cultures, which fail to recapitulate spatial aspects of cell-to-cell interactions as well as tissue gradients present in three-dimensional tumours. Here, we describe an engineered model to assemble three-dimensional tumours by rolling a scaffold–tumour composite strip. By unrolling the strip, the model can be rapidly disassembled for snapshot analysis, allowing spatial mapping of cell metabolism in concert with cell phenotype. We also show that the establishment of oxygen gradients within samples that are shaped by oxygen-dependent signalling pathways, as well as the consequential variations in cell growth, response to hypoxic gradients extending from normoxia to severe hypoxia, and therapy responsiveness, are consistent with those of tumours in vivo. Moreover, by using liquid chromatography tandem mass spectrometry, we mapped cellular metabolism and identified spatially defined metabolic signatures of cancer cells to reveal both known and novel metabolic responses to hypoxia.

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Figure 1: Schematic of TRACER design concept.
Figure 2: Cellular behaviour within the TRACER.
Figure 3: Oxygen gradients within the TRACER over time.
Figure 4: Layer-specific metabolomic analysis and correlation with hypoxia in the TRACER.
Figure 5: Metabolites with significant correlations with level of EF5 binding for shGFP and shHIF cells.

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Change history

  • 01 December 2015

    Corrected online 1 December 2015. In the version of the Article originally published online, in Fig. 1 there were some image display errors in panels a and b and the label 'GFP SK-OV-3' should have been green in panels c and e. In Fig. 2, panels a and b, layer numbers should have been defined. These errors have now been corrected in all versions of the Article.

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Acknowledgements

The authors acknowledge V. Bindokas (University of Chicago), M. Macasaet-Peralta (Pathology Research Program, UHN), J. Cathcart (AOMF), J. Stewart and D. Scollard (STTARR), B. Calvieri, S. Doyle, J. Soleas, S. Javaherian, C. Londono, C. Crossman, R. Vellanki, J. Han, M. Young and S. Lakhani (University of Toronto) for technical assistance. This work was funded by a Natural Science and Engineering Council Discovery Accelerator Supplement (RGPIN-314056) to A.P.M., a YSF NSERC fellowship to D.R., MRC Cancer Unit Core Funding to C.F. and E.G., Ontario Ministry of Health and Long Term Care (OMOHLTC), the Terry Fox New Frontiers Research Program (PPG09-020005) and the Canadian Institute for Health Research (CIHR grant 201592) grants to B.G.W., and by a Ontario Graduate Scholarship to D.C.

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Author notes

  1. Christian Frezza and Bradly G. Wouters: Co-secondary senior authors.

Authors and Affiliations

  1. Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street Toronto, Ontario M5S 3E5, Canada

    Darren Rodenhizer, Radhakrishnan Mahadevan & Alison P. McGuigan

  2. MRC Cancer Unit, University of Cambridge, Hutchison/MRC Research Centre, Box 197, Cambridge Biomedical Campus, Cambridge CB2 0XZ, UK

    Edoardo Gaude & Christian Frezza

  3. Departments of Medical Biophysics and Radiation Oncology, Princess Margaret Cancer Centre and Campbell Family Institute for Cancer Research, University Health Network, Toronto, Ontario M5G 2M9, Canada

    Dan Cojocari & Bradly G. Wouters

  4. Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario M5S 3G9, Canada

    Radhakrishnan Mahadevan & Alison P. McGuigan

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Contributions

D.R., E.G., D.C., R.M., B.G.W., C.F. and A.P.M. designed experiments, analysed the data and wrote the manuscript. D.R., E.G. and D.C. conducted experiments.

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Correspondence to Alison P. McGuigan.

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The authors declare no competing financial interests.

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Rodenhizer, D., Gaude, E., Cojocari, D. et al. A three-dimensional engineered tumour for spatial snapshot analysis of cell metabolism and phenotype in hypoxic gradients. Nature Mater 15, 227–234 (2016). https://doi.org/10.1038/nmat4482

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