Negative Regulation-Resistant p53 Variant Enhances Oncolytic Adenoviral Gene Therapy

Abstract

Intact p53 function is essential for responsiveness to cancer therapy. However, p53 activity is attenuated by the proto-oncoprotein Mdm2, the adenovirus protein E1B 55kD, and the p53 C-terminal domain. To confer resistance to Mdm2, E1B 55kD, and C-terminal negative regulation, we generated a p53 variant (p53VP Delta 30) by deleting the N-terminal and C-terminal regions of wild-type p53 and inserting the transcriptional activation domain of herpes simplex virus VP16 protein. The oncolytic adenovirus vector Ad-m Delta 19 expressing p53VP Delta 30 (Ad-m Delta 19/p53VP Delta 30) showed greater cytotoxicity than Ad-m Delta 19 expressing wild-type p53 or other p53 variants in human cancer cell lines. We found that Ad-m Delta 19/p53VP Delta 30 induced apoptosis through accumulation of p53VP Delta 30, regardless of endogenous p53 and Mdm2 status. Moreover, Ad-m Delta 19/p53VP Delta 30 showed a greater antitumor effect and increased survival rates of mice with U343 brain cancer xenografts that expressed wild-type p53 and high Mdm2 levels. To our knowledge, this is the first study reporting a p53 variant modified at the N terminus and C terminus that shows resistance to degradation by Mdm2 and E1B 55kD, as well as negative regulation by the p53 C terminus, without decreased trans-activation activity. Taken together, these results indicate that Ad-m Delta 19/p53VP Delta 30 shows potential for improving p53-mediated cancer gene therapy.