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

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.

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).

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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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