Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

In vivo imaging of lymph node metastasis with telomerase-specific replication-selective adenovirus

Nature Medicine volume 12, pages 1213–1219 (2006)Cite this article

Abstract

Currently available methods for detection of tumors in vivo such as computed tomography and magnetic resonance imaging are not specific for tumors. Here we describe a new approach for visualizing tumors whose fluorescence can be detected using telomerase-specific replication-competent adenovirus expressing green fluorescent protein (GFP) (OBP-401). OBP-401 contains the replication cassette, in which the human telomerase reverse transcriptase (hTERT) promoter drives expression of E1 genes, and the GFP gene for monitoring viral replication. When OBP-401 was intratumorally injected into HT29 tumors orthotopically implanted into the rectum in BALB/c nu/nu mice, para-aortic lymph node metastasis could be visualized at laparotomy under a three-chip color cooled charged-coupled device camera. Our results indicate that OBP-401 causes viral spread into the regional lymphatic area and selectively replicates in neoplastic lesions, resulting in GFP expression in metastatic lymph nodes. This technology is adaptable to detect lymph node metastasis in vivo as a preclinical model of surgical navigation.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: hTERT expression and selective replication of OBP-401 in human cancer cells.
Figure 2: Selective visualization of human cancer cells in vitro.
Figure 3: Selective visualization of subcutaneous tumors in vivo.
Figure 4: Orthotopic xenografts of human colorectal cancer cells and selective visualization of lymph node metastasis in two representative mice (no.1 and no. 2).

Similar content being viewed by others

References

  1. Tearney, G.J. et al. In vivo endoscopic optical biopsy with optical coherence tomography. Science 276, 2037–2039 (1997).

    Article  CAS  Google Scholar 

  2. MacDonald, S.L. & Hansell, D.M. Staging of non-small cell lung cancer: imaging of intrathoracic disease. Eur. J. Radiol. 45, 18–30 (2003).

    Article  Google Scholar 

  3. Kelloff, G.J. et al. Progress and promise of FDG-PET imaging for cancer patient management and oncologic drug development. Clin. Cancer Res. 11, 2785–2808 (2005).

    Article  CAS  Google Scholar 

  4. McMasters, K.M. et al. Sentinel lymph node biopsy for melanoma: controversy despite widespread agreement. J. Clin. Oncol. 19, 2851–2855 (2001).

    Article  CAS  Google Scholar 

  5. Kuerer, H.M. & Newman, L.A. Lymphatic mapping and sentinel lymph node biopsy for breast cancer: developments and resolving controversies. J. Clin. Oncol. 23, 1698–1705 (2005).

    Article  Google Scholar 

  6. Koopal, S.A. et al. Frozen section analysis of sentinel lymph nodes in melanoma patients. Cancer 89, 1720–1725 (2000).

    Article  CAS  Google Scholar 

  7. Tanis, P.J. et al. Frozen section investigation of the sentinel node in malignant melanoma and breast cancer. Ann. Surg. Oncol. 8, 222–226 (2001).

    Article  CAS  Google Scholar 

  8. Gulec, S.A., Su, J., O'Leary, J.P. & Stolier, A. Clinical utility of frozen section in sentinel node biopsy in breast cancer. Am. Surg. 67, 529–532 (2001).

    CAS  PubMed  Google Scholar 

  9. Chao, C. et al. Utility of intraoperative frozen section analysis of sentinel lymph nodes in breast cancer. Am. J. Surg. 182, 609–615 (2001).

    Article  CAS  Google Scholar 

  10. Misteli, T. & Spector, D.L. Applications of the green fluorescent protein in cell biology and biotechnology. Nat. Biotechnol. 15, 961–964 (1997).

    Article  CAS  Google Scholar 

  11. van Roessel, P. & Brand, A.H. Imaging into the future: visualizing gene expression and protein interactions with fluorescent proteins. Nat. Cell Biol. 4, E15–E20 (2002).

    Article  CAS  Google Scholar 

  12. Ehrhardt, D. GFP technology for live cell imaging. Curr. Opin. Plant Biol. 6, 622–628 (2003).

    Article  CAS  Google Scholar 

  13. Yang, M., Baranov, E., Moossa, A.R., Penman, S. & Hoffman, R.M. Visualizing gene expression by whole-body fluorescence imaging. Proc. Natl Acad. Sci. USA 97, 12278–12282 (2000).

    Article  CAS  Google Scholar 

  14. Hasegawa, S. et al. In vivo tumor delivery of the green fluorescent protein gene to report future occurrence of metastasis. Cancer Gene Ther. 7, 1336–1340 (2000).

    Article  CAS  Google Scholar 

  15. Ohtani, S. et al. Quantitative analysis of p53-targeted gene expression and visualization of p53 transcriptional activity following intratumoral administration of adenoviral p53 in vivo. Mol. Cancer Ther. 3, 93–100 (2004).

    CAS  PubMed  Google Scholar 

  16. Kawashima, T. et al. Telomerase-specific replication-selective virotherapy for human cancer. Clin. Cancer Res. 10, 285–292 (2004).

    Article  CAS  Google Scholar 

  17. Taki, M. et al. Enhanced oncolysis by a tropism-modified telomerase-specific replication selective adenoviral agent OBP-405 (“Telomelysin-RGD”). Oncogene 24, 3130–3140 (2005).

    Article  CAS  Google Scholar 

  18. Umeoka, T. et al. Visualization of intrathoracically disseminated solid tumors in mice with optical imaging by telomerase-specific amplification of transferred green fluorescent protein gene. Cancer Res. 64, 6259–6265 (2004).

    Article  CAS  Google Scholar 

  19. Gu, J., Andreeff, M., Roth, J.A. & Fang, B. hTERT promoter induces tumor-specific Bax gene expression and cell killing in syngenic mouse tumor model and prevents systemic toxicity. Gene Ther. 9, 30–37 (2002).

    Article  CAS  Google Scholar 

  20. Tsutsumi, S., Kuwano, H., Morinaga, N., Shimura, T. & Asao, T. Animal model of para-aortic lymph node metastasis. Cancer Lett. 169, 77–85 (2001).

    Article  CAS  Google Scholar 

  21. Maehara, Y. et al. Clinical significance of occult micrometastasis in lymph nodes from patients with early gastric cancer who died of recurrence. Surgery 119, 397–402 (1996).

    Article  CAS  Google Scholar 

  22. Coello, M.C., Luketich, J.D., Litle, V.R. & Godfrey, T.E. Prognostic significance of micrometastasis in non-small-cell lung cancer. Clin. Lung Cancer 5, 214–225 (2004).

    Article  Google Scholar 

  23. Fujiwara, T. et al. Multicenter phase I study of repeated intratumoral delivery of adenoviral p53 (ADVEXIN) in patients with advanced non-small cell lung cancer. J. Clin. Oncol. 24, 1689–1699 (2006).

    Article  CAS  Google Scholar 

  24. Hiyama, K. et al. Activation of telomerase in human lymphocytes and hematopoietic progenitor cells. J. Immunol. 155, 3711–3715 (1995).

    CAS  PubMed  Google Scholar 

  25. Adams, J.Y. et al. Visualization of advanced human prostate cancer lesions in living mice by a targeted gene transfer vector and optical imaging. Nat. Med. 8, 891–897 (2002).

    Article  CAS  Google Scholar 

  26. Yang, M., Luiken, G., Baranov, E. & Hoffman, R.M. Facile whole-body imaging of internal fluorescent tumors in mice with an LED flashlight. Biotechniques 39, 170–172 (2005).

    Article  CAS  Google Scholar 

  27. Watanabe, T. et al. Histone deacetylase inhibitor FR901228 enhances the antitumor effect of telomerase-specific replication-selective adenoviral agent OBP-301 in human lung cancer cells. Exp. Cell Res. 312, 256–265 (2006).

    CAS  PubMed  Google Scholar 

  28. Fujiwara, T. et al. Enhanced antitumor efficacy of telomerase-selective oncolytic adenoviral agent OBP-401 with docetaxel: preclinical evaluation of chemovirotherapy. Int. J. Cancer 119, 432–440 (2006).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (T.F. and S.K.); and by grants from the Ministry of Health, Labour, and Welfare of Japan (T.F.). We thank K. Nagai and H. Kawamura for the helpful discussion, and Y. Shirakiya and N. Mukai for the excellent technical support.

Author information

Authors and Affiliations

  1. Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan

    Hiroyuki Kishimoto, Toru Kojima, Yuichi Watanabe, Shunsuke Kagawa, Toshiya Fujiwara, Futoshi Uno, Fuminori Teraishi, Noriaki Tanaka & Toshiyoshi Fujiwara

  2. Center for Gene and Cell Therapy, Okayama University Hospital, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan

    Hiroyuki Kishimoto, Toru Kojima, Shunsuke Kagawa, Toshiya Fujiwara, Futoshi Uno, Fuminori Teraishi & Toshiyoshi Fujiwara

  3. Oncolys BioPharma, Inc., 3-16-33 Roppongi, Minato-ku, Tokyo, 106-0032, Japan

    Yuichi Watanabe, Yuuri Hashimoto & Yasuo Urata

  4. Department of Obstetrics and Gynecology, Kanazawa University School of Medicine, 13-1 Takara-machi, Kanazawa, 920-8641, Japan

    Satoru Kyo

  5. Laboratory of Gene Transfer and Regulation, National Institute of Biomedical Innovation, 7-6-8 Saito-Asagi, Ibaraki, 567-0085, Osaka, Japan

    Hiroyuki Mizuguchi

Authors
  1. Hiroyuki Kishimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Toru Kojima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuichi Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shunsuke Kagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Toshiya Fujiwara
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Futoshi Uno
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fuminori Teraishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Satoru Kyo
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hiroyuki Mizuguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yuuri Hashimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yasuo Urata
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Noriaki Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Toshiyoshi Fujiwara
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

T.F. conceived the idea for this project, designed all experiments and wrote the manuscript. H.K., Y.W. and Y.H. performed all laboratory experiments and H.K., T.K. and Y.W. performed all animal experiments. S.K. provided crucial ideas and helped with data interpretation. T.F., F.U., F.T. and N.T. provided technical assistance. S.K. provided the hTERT promoter. H.M. constructed the OBP-401 virus. Y.U. developed a protocol for virus manufacture and validation.

Corresponding author

Correspondence to Toshiyoshi Fujiwara.

Ethics declarations

Competing interests

Y.U. is Chief Executive Officer and T.F. is Chief Scientific Officer of Oncolys BioPharma, Inc. N.T. is Chief of the Scientific Advisory Board of Oncolys BioPharma, Inc. Y.W. and Y.H. are employees of Oncolys BioPharma, Inc. T.F. and N.T. are inventors designated on the patent application for OBP-401.

Supplementary information

Supplementary Fig. 1

In vitro and in vivo oncolytic effect of OBP-401. (PDF 28 kb)

Supplementary Fig. 2

Selective replication of OBP-401 and visualization of mouse cancer cells in vitro and in vivo. (PDF 77 kb)

Supplementary Fig. 3

Immunohistochemical analysis of GFP protein expression in metastatic lymph nodes. (PDF 43 kb)

Supplementary Fig. 4

OBP-401 replication in inflammatory lesions. (PDF 39 kb)

Supplementary Table 1

Expression of hTERT protein in lymph nodes of patients with gastric cancer. (PDF 20 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kishimoto, H., Kojima, T., Watanabe, Y. et al. In vivo imaging of lymph node metastasis with telomerase-specific replication-selective adenovirus. Nat Med 12, 1213–1219 (2006). https://doi.org/10.1038/nm1404

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm1404

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing