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

An Erratum to this article was published on 01 May 2008

This article has been updated


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.


  1. Vogelstein, B. et al. Genetic alterations during colorectal-tumor development. N. Engl. J. Med. 319, 525–532 (1988).

    Article  CAS  Google Scholar 

  2. Winawer, S.J. et al. Prevention of colorectal cancer by colonoscopic polypectomy. The National Polyp Study Workgroup. N. Engl. J. Med. 329, 1977–1981 (1993).

    Article  CAS  Google Scholar 

  3. American Cancer Society. Cancer Facts & Figures 2007 (American Cancer Society, Atlanta, 2006).

  4. Kudo, S. et al. Colonoscopic diagnosis and management of nonpolypoid early colorectal cancer. World J. Surg. 24, 1081–1090 (2000).

    Article  CAS  Google Scholar 

  5. Mayer, R., Wong, W.D., Rothenberger, D.A., Goldberg, S.M. & Madoff, R.D. Colorectal cancer in inflammatory bowel disease: a continuing problem. Dis. Colon Rectum 42, 343–347 (1999).

    Article  CAS  Google Scholar 

  6. Garcea, G. et al. Molecular biomarkers of colorectal carcinogenesis and their role in surveillance and early intervention. Eur. J. Cancer 39, 1041–1052 (2003).

    Article  CAS  Google Scholar 

  7. Goldsmith, S.J. Receptor imaging: competitive or complementary to antibody imaging? Semin. Nucl. Med. 27, 85–93 (1997).

    Article  CAS  Google Scholar 

  8. Scott, J.K. & Smith, G.P. Searching for peptide ligands with an epitope library. Science 249, 386–390 (1990).

    Article  CAS  Google Scholar 

  9. Kelly, K.A. & Jones, D.A. Isolation of a colon tumor specific binding peptide using phage display selection. Neoplasia 5, 437–444 (2003).

    Article  CAS  Google Scholar 

  10. Abraham, J.M. et al. Novel decapeptides that bind avidly and deliver radioisotope to colon cancer cells. PLoS ONE 2, e964 (2007).

    Article  Google Scholar 

  11. Kelly, K., Alencar, H., Funovics, M., Mahmood, U. & Weissleder, R. Detection of invasive colon cancer using a novel, targeted, library-derived fluorescent peptide. Cancer Res. 64, 6247–6251 (2004).

    Article  CAS  Google Scholar 

  12. Zangani, D. et al. Multiple differentiation pathways of rat mammary stromal cells in vitro: acquisition of a fibroblast, adipocyte or endothelial phenotype is dependent on hormonal and extracellular matrix stimulation. Differentiation 64, 91–101 (1999).

    Article  CAS  Google Scholar 

  13. Walters, R.W. et al. Basolateral localization of fiber receptors limits adenovirus infection from the apical surface of airway epithelia. J. Biol. Chem. 274, 10219–10226 (1999).

    Article  CAS  Google Scholar 

  14. Pasqualini, R. & Ruoslahti, E. Organ targeting in vivo using phage display peptide libraries. Nature 380, 364–366 (1996).

    Article  CAS  Google Scholar 

  15. Carson-Walter, E.B. et al. Cell surface tumor endothelial markers are conserved in mice and humans. Cancer Res. 61, 6649–6655 (2001).

    CAS  PubMed  Google Scholar 

  16. Arap, W. et al. Steps toward mapping the human vasculature by phage display. Nat. Med. 8, 121–127 (2002).

    Article  CAS  Google Scholar 

  17. Wang, T.D. et al. Functional imaging of colonic mucosa with a fibered confocal microscope for real-time in vivo pathology. Clin. Gastroenterol. Hepatol. 5, 1300–1305 (2007).

    Article  Google Scholar 

  18. Fujimoto, J.G. Optical coherence tomography for ultrahigh resolution in vivo imaging. Nat. Biotechnol. 21, 1361–1367 (2003).

    Article  CAS  Google Scholar 

  19. Wang, T.D. et al. Fluorescence endoscopic imaging of human colonic adenomas. Gastroenterology 111, 1182–1191 (1996).

    Article  CAS  Google Scholar 

  20. Haringsma, J. et al. Autofluorescence endoscopy: feasibility of detection of GI neoplasms unapparent to white light endoscopy with an evolving technology. Gastrointest. Endosc. 53, 642–650 (2001).

    Article  CAS  Google Scholar 

  21. Machida, H. et al. Narrow-band imaging in the diagnosis of colorectal mucosal lesions: a pilot study. Endoscopy 36, 1094–1098 (2004).

    Article  CAS  Google Scholar 

  22. Messmann, H., Knuchel, R., Baumler, W., Holstege, A. & Scholmerich, J. Endoscopic fluorescence detection of dysplasia in patients with Barrett's esophagus, ulcerative colitis, or adenomatous polyps after 5-aminolevulinic acid-induced protoporphyrin IX sensitization. Gastrointest. Endosc. 49, 97–101 (1999).

    Article  CAS  Google Scholar 

  23. Folli, S. et al. Immunophotodiagnosis of colon carcinomas in patients injected with fluoresceinated chimeric antibodies against carcinoembryonic antigen. Proc. Natl. Acad. Sci. USA 89, 7973–7977 (1992).

    Article  CAS  Google Scholar 

  24. Keller, R., Winde, G., Terpe, H.J., Foerster, E.C. & Domschke, W. Fluorescence endoscopy using a fluorescein-labeled monoclonal antibody against carcinoembryonic antigen in patients with colorectal carcinoma and adenoma. Endoscopy 34, 801–807 (2002).

    Article  CAS  Google Scholar 

  25. Kiesslich, R. et al. Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo. Gastroenterology 127, 706–713 (2004).

    Article  Google Scholar 

  26. Ho, S.B. et al. Heterogeneity of mucin gene expression in normal and neoplastic tissues. Cancer Res. 53, 641–651 (1993).

    CAS  PubMed  Google Scholar 

  27. Moore, A., Medarova, Z., Potthast, A. & Dai, G. In vivo targeting of underglycosylated MUC-1 tumor antigen using a multimodal imaging probe. Cancer Res. 64, 1821–1827 (2004).

    Article  CAS  Google Scholar 

  28. Soler, A.P. et al. Increased tight junctional permeability is associated with the development of colon cancer. Carcinogenesis 20, 1425–1431 (1999).

    Article  CAS  Google Scholar 

  29. Peles, E. et al. Identification of a novel contactin-associated transmembrane receptor with multiple domains implicated in protein-protein interactions. EMBO J. 16, 978–988 (1997).

    Article  CAS  Google Scholar 

  30. Su, J.L. et al. Knockdown of contactin-1 expression suppresses invasion and metastasis of lung adenocarcinoma. Cancer Res. 66, 2553–2561 (2006).

    Article  CAS  Google Scholar 

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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



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).

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