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

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.

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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).

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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

Authors

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)

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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

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

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