Optimization of antitumor efficacy and safety of in vivo cytokine gene therapy using RGD fiber-mutant adenovirus vector for preexisting murine melanoma.

We previously reported that RGD fiber-mutant adenovirus vector (AdRGD) was a very useful vector system for in vivo cytokine gene therapy for established murine B16BL6 melanoma. However, intratumoral administration of AdRGD expressing tumor necrosis factor alpha (AdRGD-TNFalpha) at high dose revealed not only the dramatic reinforcement of anti-tumor effect but also serious adverse effects, such as body weight reduction and sudden death, caused by high-level TNF-alpha leakage from the tumor into circulation. These results strongly suggested that the determination of 'limiting dose', which demonstrated therapeutic effectiveness without adverse effect, against each vector was important for the development of appropriate cytokine gene therapy. In the present study, we investigated the efficacy and the safety of AdRGD expressing interleukin-12 (AdRGD-IL12) in murine melanoma model, and determined its limiting dose. Moreover, we demonstrated that combination therapy using AdRGD-IL12 and AdRGD-TNFalpha at limiting doses or less could achieve more effective tumor regression without adverse effects. Therefore, we conclude that a combination of multiple AdRGD expressing cytokines having distinct anti-tumor mechanisms can contribute to the establishment of in vivo cytokine gene therapy for melanoma, which possesses both excellent efficacy and high safety.