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Author
Umeoka, Tatsuo
Kawashima, Takeshi
Teraishi, Fuminori
Taki, Masaki
Nishizaki, Masahiko
Kyo, Satoru
Nagai, Katsuyuki
Urata, Yasuo
Tanaka, Noriaki
Abstract
Currently available methods for detection of tumors in vivo such as X-ray, computed tomography, and ultrasonography are noninvasive and have been well studied; the images, however, are not specific for tumors. Direct optical imaging of tumor cells in vivo that can clearly distinguish them from surrounding normal tissues may be clinically useful. Here, we describe a new approach to visualizing tumors whose fluorescence can be detected using tumor-specific replication-competent adenovirus (OBP-301, Telomelysin) in combination with Ad-GFP, a replication-deficient adenovirus expressing green fluorescent protein (GFP). Human telomerase reverse transcriptase is the catalytic subunit of telomerase, which is highly active in cancer cells but quiescent in most normal somatic cells. We constructed an adenovirus 5 vector in which the human telomerase reverse transcriptase promoter element drives expression of E1A and E1B genes linked with an internal ribosome entry site and showed that OBP-301 replicated efficiently in human cancer cells, but not in normal cells such as human fibroblasts. When the human lung and colon cancer cell lines were infected with Ad-GFP at a low multiplicity of infection, GFP expression could not be detected under a fluorescence microscope; in the presence of OBP-301, however, Ad-GFP replicated in these tumor cells and showed strong green signals. In contrast, coinfection with OBP-301 and Ad-GFP did not show any signals in normal cells such as fibroblasts and vascular endothelial cells. We also found that established subcutaneous tumors could be visualized after intraturnoral injection of OBP-301 and Ad-GFP. A549 human lung tumors and SW620 human colon tumors transplanted into BALB/c nu/nu mice were intraturnorally injected with 8 X 10(5) plaque-forming units of Ad-GFP in combination with 8 X 106 plaque-forming units of OBP-301. Within 3 days of treatment, the fluorescence of the expressed GFP became visible by a three-chip color cooled charged-coupled device camera in these tumors, whereas intraturnoral injection of Ad-GFP alone could not induce GFP fluorescence. Moreover, intrathoracic administration of Ad-GFP and OBP-301 could visualize disseminated A549 tumor nodules in mice after intrathoracic implantation. Our results indicate that intratumoral or intrathoracic injection of Ad-GFP in combination with OBP-301 might be a useful diagnostic method that provides a foundation for future clinical application.
Keywords
GFP
adenovirus
telomerase
replication
gene therapy
Note
Published with permission from the copyright holder. This is the author's copy, as published in Cancer Research, September 2004, Volume 64, Issue 17, Pages 6259-6265.
Direct access to Thomson Web of Science record
Copyright © 2004 American Association for Cancer Research. All rights reserved.
Published Date
2004-09-01
Publication Title
Cancer Research
Volume
volume64
Issue
issue17
Publisher
American Association for Cancer Research
Start Page
6259
End Page
6265
ISSN
0008-5472
NCID
AA00598557
Content Type
Journal Article
Official Url
http://cancerres.aacrjournals.org/content/64/17/6259
language
英語
Copyright Holders
American Association for Cancer Research
File Version
author
Refereed
True
PubMed ID
Web of Sience KeyUT
Submission Path
oncology/4