Abstract

Profiling the heterogeneous phenotypes of rare circulating tumour cells (CTCs) in whole blood is critical to unravelling the complex and dynamic properties of these potential clinical markers. This task is challenging because these cells are present at parts per billion levels among normal blood cells. Here we report a new nanoparticle-enabled method for CTC characterization, called magnetic ranking cytometry, which profiles CTCs on the basis of their surface expression phenotype. We achieve this using a microfluidic chip that successfully processes whole blood samples. The approach classifies CTCs with single-cell resolution in accordance with their expression of phenotypic surface markers, which is read out using magnetic nanoparticles. We deploy this new technique to reveal the dynamic phenotypes of CTCs in unprocessed blood from mice as a function of tumour growth and aggressiveness. We also test magnetic ranking cytometry using blood samples collected from cancer patients.

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References

  1. 1.

    & A perspective on cancer cell metastasis. Science 331, 1559–1564 (2011).

  2. 2.

    , & Circulating tumor cells. Science 341, 1186–1188 (2013).

  3. 3.

    & Challenges in circulating tumour cell research. Nat. Rev. Cancer 14, 623–631 (2014).

  4. 4.

    , & Circulating tumor cells: getting more from less. Sci. Transl. Med. 4, 141ps13 (2012).

  5. 5.

    et al. Beyond the capture of circulating tumor cells: next-generation devices and materials. Angew. Chem. Int. Ed. 55, 1252–1265 (2016).

  6. 6.

    et al. Marker-specific sorting of rare cells using dielectrophoresis. Proc. Natl Acad. Sci. USA 102, 15757–15761 (2005).

  7. 7.

    et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature 450, 1235–1239 (2007).

  8. 8.

    et al. Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor. J. Am. Chem. Soc. 130, 8633–8641 (2008).

  9. 9.

    et al. Isolating highly enriched populations of circulating epithelial cells and other rare cells from blood using a magnetic sweeper device. Proc. Natl Acad. Sci. USA 106, 3970–3975 (2009).

  10. 10.

    et al. Isolation of circulating tumor cells using a microvortex-generating herringbone-chip. Proc. Natl Acad. Sci. USA 107, 18392–18397 (2010).

  11. 11.

    et al. Highly efficient capture of circulating tumor cells by using nanostructured silicon substrates with integrated chaotic micromixers. Angew. Chem. Int. Ed. 50, 3084–3088 (2011).

  12. 12.

    et al. Sensitive and high-throughput isolation of rare cells from peripheral blood with ensemble-decision aliquot ranking. Angew. Chem. Int. Ed. 51, 4618–4622 (2012).

  13. 13.

    et al. Bioinspired multivalent DNA network for capture and release of cells. Proc. Natl Acad. Sci. USA 109, 19626–19631 (2012).

  14. 14.

    et al. Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells. Sci. Transl. Med. 5, 179ra47 (2013).

  15. 15.

    et al. Tunable nanostructured coating for the capture and selective release of viable circulating tumor cells. Adv. Mater. 27, 1593–1599 (2015).

  16. 16.

    , , , & Discontinuous nanoporous membranes reduce non-specific fouling for immunoaffinity cell capture. Small 9, 4207–4214 (2013).

  17. 17.

    et al. A novel 3D integrated platform for the high-resolution study of cell migration plasticity. Macromol. Biosci. 13, 973–983 (2013).

  18. 18.

    et al. Nanoparticle-mediated binning and profiling of heterogeneous circulating tumor cell subpopulations. Angew. Chem. Int. Ed. 54, 139–143 (2015).

  19. 19.

    et al. Genetic analysis of H1N1 influenza virus from throat swab samples in a microfluidic system for point-of-care diagnostics. J. Am. Chem. Soc. 133, 9129–9135 (2011).

  20. 20.

    , , & Microscale magnetic field modulation for enhanced capture and distribution of rare circulating tumor cells. Sci. Rep. 5, 8745 (2015).

  21. 21.

    et al. Immune-induced epithelial to mesenchymal transition in vivo generates breast cancer stem cells. Cancer Res. 69, 2887–2895 (2009).

  22. 22.

    et al. Circulating breast tumor cells exhibit dynamic changes in epithelial and mesenchymal composition. Science 339, 580–584 (2013).

  23. 23.

    , , , & Translational applications of flow cytometry in clinical practice. J. Immunol. 188, 4715–4719 (2012).

  24. 24.

    , , & Prognostic significance of Gleason score 3+4 versus Gleason score 4+3 tumor at radical prostatectomy. Urology 56, 823–827 (2000).

  25. 25.

    et al. Ultrasensitive clinical enumeration of rare cells ex vivo using a micro-hall detector. Sci. Transl. Med. 4, 141ra192 (2012).

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Acknowledgements

The authors acknowledge generous support from the Canadian Institutes of Health Research (Emerging Team grant, POP grant), the Ontario Research Fund (ORF Research Excellence grant), the Canadian Cancer Society Research Institute (Innovation grant) and the Connaught Foundation. We also acknowledge all of the patients and healthy donors who donated specimens to our studies.

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Affiliations

  1. Department of Electrical and Computer Engineering, Faculty of Engineering, University of Toronto, Toronto, Ontario M5S 3G4, Canada

    • Mahla Poudineh
    •  & Edward H. Sargent
  2. Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario M5S 3M2, Canada

    • Peter M. Aldridge
    • , Brenda J. Green
    • , Leyla Kermanshah
    •  & Shana O. Kelley
  3. Department of Pharmaceutical Science, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada

    • Sharif Ahmed
    • , Vivian Nguyen
    • , Carmen Tu
    • , Reza M. Mohamadi
    •  & Shana O. Kelley
  4. Sunnybrook Health Sciences Centre, Sunnybrook Research Institute, University of Toronto, Toronto, Ontario M4N 3M5, Canada

    • Robert K. Nam
  5. Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Ontario M5G 2M9, Canada

    • Aaron Hansen
    • , Srikala S. Sridhar
    • , Antonio Finelli
    • , Neil E. Fleshner
    •  & Anthony M. Joshua
  6. Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada

    • Shana O. Kelley

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Contributions

M.P., P.M.A., S.A., B.J.G., L.K., V.N., C.T., R.M.M., S.O.K. and E.H.S. conceived and designed the experiments. M.P., P.M.A., S.A. B.J.G., L.K., V.N., C.T. and R.M.M. performed the experiments and analysed the data. R.K.N., A.H., S.S.S., A.F., N.E.F. and A.M.J. contributed clinical expertise and clinical specimens. All authors discussed the results and contributed to the preparation and editing of the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Edward H. Sargent or Shana O. Kelley.

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DOI

https://doi.org/10.1038/nnano.2016.239

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