Abstract 4065: A simple biological imaging system for detecting adenovirus receptor expression in tumor cells using a telomerase-specific replication-competent adenovirus

Abstract

Abstract Adenovirus serotype 5 (Ad5) is frequently used as an effective vector for induction of therapeutic transgenes in cancer gene therapy or of tumor cell lysis in oncolytic virotherapy. Ad5 can infect target cells through binding with the coxsackie and adenovirus receptor (CAR). Thus, the infectious ability of Ad5-based vectors depends on the CAR expression level in target cells. There are conventional methods to evaluate the CAR expression level in human target cells, including flow cytometry, western blotting and immunocytochemistry. We recently generated a green fluorescent protein (GFP)-expressing telomerase-specific replication-competent adenovirus OBP-401, which induces ectopic GFP expression in tumor cells, but not in normal cells. In this study, we evaluated whether induction of GFP expression by OBP-401 infection is associated with CAR expression in tumor cells. OBP-401-mediated GFP induction was further examined in xenograft tumor tissues that have different levels of CAR expression and in surrounding normal tissues. We used six human sarcoma cell lines that have different levels of CAR expression and CAR-positive normal human lung fibroblasts (NHLF). Flow cytometry analysis confirmed that cells of four of the sarcoma cell lines (OST, U2OS, NOS-10 and MNNG/HOS) as well as the NHLF cells showed detectable CAR expression, whereas cells of the NMFH-1 and OUMS-27 sarcoma cell lines had no detectable CAR expression. To determine suitable conditions for OBP-401 infection, all tumor and normal cells was infected with OBP-401 at multiplicity of infections of 1, 10 and 100 plaque forming units/cell. OBP-401 infection induced detectable GFP expression in CAR-expressing tumor cells, but not in CAR-negative tumor cells, nor in CAR-positive normal fibroblasts, 24 hours after infection. OBP-401-mediated GFP expression was significantly associated with CAR expression in tumor cells. OBP-401 infection detected tumor cells with low CAR expression more efficiently than conventional methods. We further examined the potential of the OBP-401-mediated method for the detection of CAR expression in tumor and normal tissues, we used this method to analyze CAR expression of human xenograft tumor tissues. OBP-401 infection induced GFP expression in OST tumor tissues, but not in OUMS-27 tumor tissues or in normal muscle tissue. OBP-401 also distinguished CAR-positive tumor tissues from CAR-negative tumor and normal tissues in biopsy samples. These results suggest that GFP-expressing telomerase-specific replication-competent adenovirus is a very potent diagnostic tool for assessment of functional CAR expression in tumor cells for Ad5-based antitumor therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4065. doi:1538-7445.AM2012-4065