Nanoplasmonic quantification of tumour-derived extracellular vesicles in plasma microsamples for diagnosis and treatment monitoring

Abstract

Tumour-derived extracellular vesicles (EVs) are of increasing interest as a resource of diagnostic biomarkers. However, most EV assays require large samples and are time-consuming, low-throughput and costly, and thus impractical for clinical use. Here, we describe a rapid, ultrasensitive and inexpensive nanoplasmon-enhanced scattering (nPES) assay that directly quantifies tumour-derived EVs from as little as 1 μl of plasma. The assay uses the binding of antibody-conjugated gold nanospheres and nanorods to EVs captured by EV-specific antibodies on a sensor chip to produce a local plasmon effect that enhances tumour-derived EV detection sensitivity and specificity. We identified a pancreatic cancer EV biomarker, ephrin type-A receptor 2 (EphA2), and demonstrate that an nPES assay for EphA2-EVs distinguishes pancreatic cancer patients from pancreatitis patients and healthy subjects. EphA2-EVs were also informative in staging tumour progression and in detecting early responses to neoadjuvant therapy, with better performance than a conventional enzyme-linked immunosorbent assay. The nPES assay can be easily refined for clinical use, and readily adapted for diagnosis and monitoring of other conditions with disease-specific EV biomarkers.

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Figure 1: Design of an nPES platform for EV detection.
Figure 2: Characterization of the performance of the nPES assay with exosome-spiked human plasma samples.
Figure 3: Identification of pancreatic-cancer-associated EV as a potential biomarker by establishing an nPES-EphA2-EV detection system.
Figure 4: EphA2-EV detection and clinical performance.

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Acknowledgements

We thank P. Mcshane at Houston Methodist Hospital and M. W. Hurd at the University of Texas M.D. Anderson Cancer Center for organizing clinical samples in this study. The work was primarily supported by research funding provided by NIH grants R01Al113725, R01AI122932, U54CA143837 and 5P50CA126752-08, an NIH intramural research program, a John S. Dunn Foundation award, a PANCAN-AACR Career Development Award (14-20-25-KOAY), a Radiological Society of North America seed grant (RSD1429), and the Sheikh Ahmed Center for Pancreatic Cancer Research and the Center for Radiation Oncology Research at The University of Texas M.D. Anderson Cancer Center.

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K.L., F.L., J.F. and Y.H. designed the research plan. K.L., F.L., J.F., D.S. and C.L. performed the experiments. E.J.K., D.W.B., K.Y. and M.H.K. collected the clinical samples and clinical data. K.L., F.L. C.J.L., Y.L., Z.Z. and Y.H. performed data analysis. K.L., F.L., C.J.L., Z.Z. and Y.H. wrote the manuscript, and all authors contributed to the revision of the manuscript.

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Correspondence to Kai Liang or Fei Liu or Ye Hu.

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The authors declare no competing interests.

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Liang, K., Liu, F., Fan, J. et al. Nanoplasmonic quantification of tumour-derived extracellular vesicles in plasma microsamples for diagnosis and treatment monitoring. Nat Biomed Eng 1, 0021 (2017). https://doi.org/10.1038/s41551-016-0021

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