Doxorubicin directs the accumulation of interleukin-12-induced IFNγ into tumors for enhancing STAT1-dependent antitumor effect

Abstract

Purpose: To examine the mechanism by which doxorubicin plus interleukin-12 (IL-12) gene transfer induces enhanced therapeutic efficacy against tumors. Experimental Design: Tumor-bearing mice were treated with doxorubicin, IL-12-encoding plasmid DNA, doxorubicin plus IL-12-encoding plasmid DNA, or plasmid DNA control. Doxorubicin was systemically given via i.p. injection, and IL-12 was systemically expressed via i.m. injection. To show that doxorubicin enhances the accumulation of IL-12-induced IFNγ into tumors and the signal transducer and activator of transcription 1 (Stat1)-dependent antitumor efficacy, the distribution of IFNγ and the therapeutic end points, such as T-cell infiltration, inhibition of tumor vessel density, tumor growth inhibition, and inhibition of spontaneous tumor metastasis in wild-type and Stat1-/- host and tumors were determined after the treatment at the indicated time points. Results: In this study, a novel mechanism was unveiled. We discovered that doxorubicin enhances the accumulation of IL-12-induced IFNγ in tumors. The doxorubicin-mediated accumulation of IFNγ in tumors is caused by an increased accumulation of IFNγ-secreting immune cells and not by a direct translocation of IFNγ protein into tumors. Depletion of immune cells reverses the doxorubicin-mediated accumulation of IFNγ into tumors and reverses the inhibition of tumor vessel density induced by coadministration of doxorubicin and IL-12 DNA. Knocking out IFNγ signaling in the tumor host reverses the significant inhibition of tumor growth by coadministration of doxorubicin and IL-12. Conclusions: The enhanced antitumor efficacy by coadministration of doxorubicin and IL-12 is dependent on the accumulation of IFNγ in tumors. This discovery provides a possible strategy to reduce side effects caused by IL-12. © 2007 American Association for Cancer Research.