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Isolation of rare circulating tumour cells in cancer patients by microchip technology

Nature volume 450, pages 1235–1239 (2007)Cite this article

Abstract

Viable tumour-derived epithelial cells (circulating tumour cells or CTCs) have been identified in peripheral blood from cancer patients and are probably the origin of intractable metastatic disease1,2,3,4. Although extremely rare, CTCs represent a potential alternative to invasive biopsies as a source of tumour tissue for the detection, characterization and monitoring of non-haematologic cancers5,6,7,8. The ability to identify, isolate, propagate and molecularly characterize CTC subpopulations could further the discovery of cancer stem cell biomarkers and expand the understanding of the biology of metastasis. Current strategies for isolating CTCs are limited to complex analytic approaches that generate very low yield and purity9. Here we describe the development of a unique microfluidic platform (the ‘CTC-chip’) capable of efficient and selective separation of viable CTCs from peripheral whole blood samples, mediated by the interaction of target CTCs with antibody (EpCAM)-coated microposts under precisely controlled laminar flow conditions, and without requisite pre-labelling or processing of samples. The CTC-chip successfully identified CTCs in the peripheral blood of patients with metastatic lung, prostate, pancreatic, breast and colon cancer in 115 of 116 (99%) samples, with a range of 5–1,281 CTCs per ml and approximately 50% purity. In addition, CTCs were isolated in 7/7 patients with early-stage prostate cancer. Given the high sensitivity and specificity of the CTC-chip, we tested its potential utility in monitoring response to anti-cancer therapy. In a small cohort of patients with metastatic cancer undergoing systemic treatment, temporal changes in CTC numbers correlated reasonably well with the clinical course of disease as measured by standard radiographic methods. Thus, the CTC-chip provides a new and effective tool for accurate identification and measurement of CTCs in patients with cancer. It has broad implications in advancing both cancer biology research and clinical cancer management, including the detection, diagnosis and monitoring of cancer10.

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Figure 1: Isolation of CTCs from whole blood using a microfluidic device.
Figure 2: CTC capture and enumeration.
Figure 3: Enumeration of CTCs from cancer patients.
Figure 4: Characterization of CTCs with tumour-specific molecular markers.

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Acknowledgements

We thank A. Amin for technical assistance in running experiments, O. Hurtado for clean room work, S. Murthy for surface chemistry, L. Romonosky for cell counting, D. Hyde for digital pictures and D. Poulsen for illustrations. We are also grateful to R. Kapur and his team for technical assistance. The authors acknowledge funding from the National Institutes of Health (to M.T.), and the Doris Duke Distinguished Clinical Scientist Award (to D.A.H.).

Author Contributions S.N., L.V.S., R.G.T., D.A.H. and M.T. designed and conducted the study. S.M., D.W.B. and L.U. performed gene expression analyses; D.I. contributed to the microfluidic system. M.R.S., E.L.K. and P.R. acquired clinical samples. U.J.B. provided input on cytopathology; A.M. performed statistical analysis; and S.D. performed radiology measurements. S.N., L.V.S., D.W.B., S.M., D.I., D.A.H. and M.T. participated in data analysis and writing of the manuscript.

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  1. Daphne W. Bell & Ulysses J. Balis

    Present address: Present addresses: National Human Genome Research Institute/NIH Cancer Genetics Branch, Bethesda, Maryland 20892, USA (D.W.B.); Department of Pathology, University of Michigan Health System, Ann Arbor, Michigan 48109, USA (U.J.B.).,

  2. Sunitha Nagrath and Lecia V. Sequist: These authors contributed equally to this paper.

Authors and Affiliations

  1. Surgical Services and BioMEMS Resource Center, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, Massachusetts 02114, USA,

    Sunitha Nagrath, Daniel Irimia, Ulysses J. Balis, Ronald G. Tompkins & Mehmet Toner

  2. Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, Massachusetts 02114, USA,

    Lecia V. Sequist, Shyamala Maheswaran, Daphne W. Bell, Lindsey Ulkus, Matthew R. Smith, Eunice L. Kwak, Subba Digumarthy, Alona Muzikansky, Paula Ryan & Daniel A. Haber

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Correspondence to Mehmet Toner.

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Competing interests

At the time the reported study was performed, M.T., R.G.T. and Massachusetts General Hospital (MGH) had significant equity holdings or similar interests in licensees from MGH for technology described in this manuscript. Before final submission for publication, recognizing Partners HealthCare System and Harvard Medical School policies on conflicts of interest, M.T., R.G.T. and MGH relinquished all such interests.

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The file contains Supplementary Figures S1-S6 with Legends and Supplementary Tables S1-S2. (PDF 852 kb)

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Nagrath, S., Sequist, L., Maheswaran, S. et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature 450, 1235–1239 (2007). https://doi.org/10.1038/nature06385

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