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Low-cost thermophoretic profiling of extracellular-vesicle surface proteins for the early detection and classification of cancers

Nature Biomedical Engineering volume 3, pages 183–193 (2019)Cite this article

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Abstract

Non-invasive assays for early cancer screening are hampered by challenges in the isolation and profiling of circulating biomarkers. Here, we show that surface proteins from serum extracellular vesicles labelled with a panel of seven fluorescent aptamers can be profiled, via thermophoretic enrichment and linear discriminant analysis, for cancer detection and classification. In a cohort of 102 patients, including 6 cancer types at stages I–IV, the assay detected stage I cancers with 95% sensitivity (95% confidence interval (CI): 74–100%) and 100% specificity (95% CI: 80–100%), and classified the cancer type with an overall accuracy of 68% (95% CI: 59–77%). For patients who underwent prostate biopsies, the assay was superior to the analysis of prostate-specific antigen levels (area under the curve: 0.94 versus 0.68; 33 patients) for the discrimination of prostate cancer and benign prostate enlargement, and also in the assessment of biochemical cancer recurrence after radical prostatectomy. The assay is inexpensive, fast, and requires small serum volumes (<1 µl), and if validated in larger cohorts may facilitate cancer screening, classification and monitoring.

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Fig. 1: Overview of TAS for the profiling of surface proteins of EVs.
Fig. 2: Characterization of thermophoretic accumulation.
Fig. 3: Characterization of TAS to profile surface proteins of EVs.
Fig. 4: EV surface protein profiles in cancer patients, as measured by TAS.
Fig. 5: Multiclass cancer diagnostics enabled by EV surface protein profiles.
Fig. 6: Detection and classification of cancers in a validation cohort.
Fig. 7: EV-based liquid biopsy in prostate cancer.

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Code availability

Custom code for linear discriminant analysis is available within the Supplementary Information.

Data availability

All data supporting the findings of this study are available within the paper and its Supplementary Information. Raw imaging data are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported financially by the NSFC (21622503 and 21475028), Youth Innovation Promotion Association of CAS (2016035) and Beijing Municipal Science and Technology Commission (Z171100001117135 and Z161100004916095).

Author information

Author notes

  1. These authors contributed equally: Chao Liu, Junxiang Zhao.

Authors and Affiliations

  1. CAS Key Laboratory of Standardization and Measurement for Nanotechnology, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, Beijing, China

    Chao Liu, Junxiang Zhao, Fei Tian, Wei Zhang, Qiang Feng, Jianqiao Chang, Baoquan Ding & Jiashu Sun

  2. University of Chinese Academy of Sciences, Beijing, China

    Chao Liu, Junxiang Zhao, Wei Zhang, Baoquan Ding & Jiashu Sun

  3. Department of Geriatric Laboratory Medicine, Chinese PLA General Hospital, Beijing, China

    Lili Cai & Yulong Cong

  4. Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, China

    Fangning Wan, Yunjie Yang & Bo Dai

  5. Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China

    Fangning Wan, Yunjie Yang & Bo Dai

  6. Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering and College of Biology, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha, China

    Weihong Tan

  7. Institute of Molecular Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Weihong Tan

  8. College of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai, China

    Weihong Tan

  9. Department of Chemistry, Center for Research at Bio/Nano Interface, UF Health Cancer Center, UF Genetics Institute, University of Florida, Gainesville, FL, USA

    Weihong Tan

  10. Department of Physiology and Functional Genomics, Center for Research at Bio/Nano Interface, UF Health Cancer Center, UF Genetics Institute, University of Florida, Gainesville, FL, USA

    Weihong Tan

Authors
  1. Chao Liu
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Contributions

J.S. and W.T. directed the research. J.S. and C.L. conceived the idea. J.Z., C.L., F.T., L.C., W.Z., Q.F., J.C., F.W. and Y.Y. performed the experiments. C.L. performed the CFD simulation and LDA algorithm. B. Dai, Y.C. and B. Ding assisted with data interpretation. All authors discussed the results and wrote the paper.

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Correspondence to Jiashu Sun.

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Liu, C., Zhao, J., Tian, F. et al. Low-cost thermophoretic profiling of extracellular-vesicle surface proteins for the early detection and classification of cancers. Nat Biomed Eng 3, 183–193 (2019). https://doi.org/10.1038/s41551-018-0343-6

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