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Detection of colonic dysplasia in vivo using a targeted heptapeptide and confocal microendoscopy

Nature Medicine volume 14, pages 454–458 (2008)Cite this article

An Erratum to this article was published on 01 May 2008

This article has been updated

Abstract

A combination of targeted probes and new imaging technologies provides a powerful set of tools with the potential to improve the early detection of cancer. To develop a probe for detecting colon cancer, we screened phage display peptide libraries against fresh human colonic adenomas for high-affinity ligands with preferential binding to premalignant tissue. We identified a specific heptapeptide sequence, VRPMPLQ, which we synthesized, conjugated with fluorescein and tested in patients undergoing colonoscopy. We imaged topically administered peptide using a fluorescence confocal microendoscope delivered through the instrument channel of a standard colonoscope. In vivo images were acquired at 12 frames per second with 50-μm working distance and 2.5-μm (transverse) and 20-μm (axial) resolution. The fluorescein-conjugated peptide bound more strongly to dysplastic colonocytes than to adjacent normal cells with 81% sensitivity and 82% specificity. This methodology represents a promising diagnostic imaging approach for the early detection of colorectal cancer and potentially of other epithelial malignancies.

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Figure 1: Binding assays for clone bearing peptide VRPMPLQ.
Figure 2: In vivo confocal fluorescence images of peptide binding.
Figure 3: In vivo confocal fluorescence images of the border between colonic adenoma and normal mucosa, showing peptide binding to dysplastic colonocytes.
Figure 4: Receiver operating characteristic (ROC) for peptide binding to adenoma versus normal colonocytes.
Figure 5: Schematic of peptide reagent development procedure.

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Change history

  • 21 March 2008

    In the version of this article initially published online, the name of the first author, Pei-Lin Hsiung, was misspelled as Pei-Lei Hsiung. The error has been corrected for all versions of the article.

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Acknowledgements

The authors acknowledge funding support from the US National Institutes of Health, including U54 CA105296 (NCI), K08 DK067618 (NIDDK) and P30 DK56339 (DDC Pilot Award), from the Doris Duke Charitable Foundation, from the Stanford School of Medicine Dean's Fellowship (P.-L.H.) and from the Cynthia Fry Gunn Research Fund. P.-L.H. is supported by the Canary Foundation/American Cancer Society Early Detection Postdoctoral Fellowship. We thank J. Kosek for support with histopathological interpretation and Mauna Kea Technologies for technical support and use of their clinical Cellvizio-GI imaging system.

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Authors and Affiliations

  1. Department of Pediatrics, Radiology and Microbiology & Immunology, Stanford University School of Medicine, 318 Campus Dr., Rm. E-150, Stanford, 94305, California, USA

    Pei-Lin Hsiung, Jonathan Hardy, Christine B Du, Amy P Wu & Christopher H Contag

  2. Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave., Bldg. 100, Palo Alto, 94304, California, USA

    Shai Friedland, Roy Soetikno, Peyman Sahbaie & Thomas D Wang

  3. Department of Medicine, Stanford University School of Medicine, 300 Pasteur Dr., Alway Bldg., Rm. M211, Stanford, 94305, California, USA

    Shai Friedland, Roy Soetikno, Anson W Lowe & Thomas D Wang

  4. Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, P.O. Box 100275, 1600 SW Archer Rd., MSB 649, Gainesville, 32610, Florida, USA

    James M Crawford

Authors
  1. Pei-Lin Hsiung
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Contributions

P.-L.H. conducted most of the experiments, with help from A.P.W. and J.H. (library clearing and cell line screening), C.B.D., P.S. and T.D.W. (clinical coordination), and T.D.W., S.F. and R.S. (in vivo studies). P.-L.H., J.H. and T.D.W. conducted the data analysis. P.-L.H., J.H., J.M.C., A.W.L., C.H.C. and T.D.W. were responsible for the concepts and writing of the manuscript. R.S., C.H.C. and T.D.W. supervised the project.

Corresponding author

Correspondence to Thomas D Wang.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–3, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 692 kb)

Supplementary Video 1

In vivo video of peptide binding to a dysplastic crypt adjacent to normal mucosa. Real time video following peptide administration shows increased binding to colonocytes of a dysplastic crypt (left half) compared to normal crypts (right half). Imaging of border provides a uniform field of view for comparison. (MOV 2737 kb)

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Hsiung, PL., Hardy, J., Friedland, S. et al. Detection of colonic dysplasia in vivo using a targeted heptapeptide and confocal microendoscopy. Nat Med 14, 454–458 (2008). https://doi.org/10.1038/nm1692

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