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Downregulation of Stat3 in melanoma: reprogramming the immune microenvironment as an anticancer therapeutic strategy

Gene Therapy volume 20, pages 1085–1092 (2013)Cite this article

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Abstract

Persistent activation of the transcription factor, signal transducer and activator of transcription 3 (Stat3) has been shown to mediate several oncogenic features in many types of cancers, including melanoma. In this study, we investigated whether lentiviral (LV) delivery of Stat3-targeting short hairpin RNA (shRNA; LV-shStat3) to K1735-C4 melanoma cells modulates antitumor immunity. Three shStat3 sequences, starting at the position 446, 830 and 1412, were cloned into a mir30 cassette. A shRNA with scrambled sequence served as a control. Transduction with LV-shStat3 resulted in downregulation of Stat3 in vitro. The latter coincided with low cell viability, a reduced expression of survivin and matrix metalloproteinase (MMP)-2. A single injection of LV-shStat3 in K1735-C4 tumors efficiently downregulated Stat3 in vivo and resulted in reduction of both vascular endothelial growth factor secretion and in myeloid-derived suppressor cell (MDSC) numbers. In contrast, we observed an increase in interleukin-6 and interferon-γ secretion, mature dendritic cells (DCs) and CD8+ T cells. Both DCs and CD8+ T cells displayed enhanced activity, whereas granulocytic MDSCs lost their suppressive capacity upon Stat3 downregulation. Importantly, a single injection of LV-shStat3 was sufficient to reduce tumor growth, hence prolong survival of tumor-bearing mice. These data demonstrate that Stat3 downregulation in melanoma reinvigorates existing antitumor immunity.

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Acknowledgements

We thank Angelo Willems and Julie Verdood for their assistance during the fluorescence-activated cell sorting analysis. We also thank Xavier Debaere, Elsy Vaeremans and Petra Roman for the plasmid DNA preparation. Funding was obtained from the Research foundation Flanders (FWO-V, Grant number #G023411N), the Agency of Innovation by Science and Technology, the Interuniversity Attraction Poles Program, the ‘Stichting tegen Kanker’ and Belgian State-Belgian Science Policy. KB is funded by the FWO-V. SM is funded by the IWT.

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  1. Department of Immunology-Physiology, Laboratory of Molecular and Cellular Therapy, Vrije Universiteit Brussel, Jette, Belgium

    P U Emeagi, S Maenhout, N Dang, C Heirman, K Thielemans & K Breckpot

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  1. P U Emeagi
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Correspondence to K Breckpot.

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Emeagi, P., Maenhout, S., Dang, N. et al. Downregulation of Stat3 in melanoma: reprogramming the immune microenvironment as an anticancer therapeutic strategy. Gene Ther 20, 1085–1092 (2013). https://doi.org/10.1038/gt.2013.35

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