This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ono, H. A. Articles by Yamamoto, M. Search for Related Content PubMed PubMed Citation Articles by Ono, H. A. Articles by Yamamoto, M.

Noninvasive Visualization of Adenovirus Replication with a Fluorescent Reporter in the E3 Region

¹ Division of Human Gene Therapy, Departments of Medicine, Pathology and Surgery, and the Gene Therapy Center, University of Alabama at Birmingham, Birmingham, Alabama and ² Department of Gastroenterological Surgery, Yokohama City University, Graduate School of Medicine, Yokohama, Japan

Requests for reprints: Masato Yamamoto, Division of Human Gene Therapy, Departments of Medicine, Pathology and Surgery, and the Gene Therapy Center, University of Alabama at Birmingham, 901 19th Street South, BMR2-410, Birmingham, AL 35294. Phone: 205-975-0172; Fax: 205-975-8565; E-mail: Masato.Yamamoto{at}ccc.uab.edu.

To overcome the inefficacy and undesirable side effects of current cancer treatment strategies, conditionally replicative adenoviruses have been developed to exploit the unique mechanism of oncolysis afforded by tumor-specific viral replication. Despite rapid translation into clinical trials and the established safety of oncolytic adenoviruses, the in vivo function of these agents is not well understood due to lack of a noninvasive detection system for adenovirus replication. To address this issue, we propose the expression of a reporter from the adenovirus E3 region as a means to monitor replication. Adenovirus replication reporter vectors were constructed with the enhanced green fluorescent protein (EGFP) gene placed in the deleted E3 region under the control of the adenoviral major late promoter while retaining expression of the adenovirus death protein to conserve the native oncolytic capability of the virus. Strong EGFP fluorescence was detected from these vectors in a replication-dependent manner, which correlated with viral DNA replication. Fluorescence imaging in vivo confirmed the ability to noninvasively detect fluorescent signal during replication, which generally corresponded with the underlying level of viral DNA replication. EGFP representation of viral replication was further confirmed by Western blot comparison with the viral DNA content in the tumors. Imaging reporter expression controlled by the adenoviral major late promoter provides a viable approach to noninvasively monitor adenovirus replication in preclinical studies and has the potential for human application with clinically relevant imaging reporters.

This article has been cited by other articles:

				J. Leyton, M. Lockley, J. L. Aerts, S. K. Baird, E. O. Aboagye, N. R. Lemoine, and I. A. McNeish Quantifying the Activity of Adenoviral E1A CR2 Deletion Mutants Using Renilla Luciferase Bioluminescence and 3'-Deoxy-3'-[18F]Fluorothymidine Positron Emission Tomography Imaging. Cancer Res., September 15, 2006; 66(18): 9178 - 9185. [Abstract] [Full Text] [PDF]