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RNA interference-mediated silencing of Foxo3 in antigen-presenting cells as a strategy for the enhancement of DNA vaccine potency

Gene Therapy volume 18, pages 372–383 (2011)Cite this article

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Abstract

The transcription factor Forkhead box O3 (Foxo3) has a critical role in suppressing the expansion of antigen-specific effector T-cell populations; hence, Foxo3 is a potential target for enhancing the antitumor immunity of cancer vaccines. In this report, we evaluated the potential of RNA interference (RNAi)-mediated silencing of Foxo3 in antigen-presenting cells as an adjuvant for HER2/neu DNA cancer vaccines. Bicistronic plasmids expressing the N-terminal extracellular domain of human HER-2/neu and the Foxo3 short hairpin RNA (hN’-neu-Foxo3 shRNA) or the scrambled control (hN’-neu-scramble shRNA) were subcutaneously injected into mice by gene gun administration to elicit antitumor immunity against p185neu-overexpressing MBT-2 bladder tumor cells. We found that mice treated with hN’-neu-Foxo3 shRNA showed greater reductions in tumor growth and longer survival times than mice treated with hN’-neu-scramble shRNA, indicating that the silencing of Foxo3 enhanced the antitumor efficacy of the HER-2/neu cancer vaccine. Cytotoxicity analyses further revealed that the Foxo3 shRNA-enhanced antitumor effect was associated with significant increases in the number of functional CD8+ T cells and in the levels of cytotoxic T lymphocytes activity. Interleukin-6 was induced by hN’-neu-Foxo3 shRNA treatment but did not have a critical role in the antitumor effect of the hN’-neu-Foxo3 shRNA vaccine. Moreover, in vivo lymphocyte depletion analyses confirmed that the antitumor efficacy of the hN’-neu-Foxo3 shRNA vaccine depended on functional CD8+ T cells. Finally, Foxo3 suppression was shown to markedly improve the effect of the HER-2/neu DNA vaccine in limiting the growth and lung metastases of MBT-2 cells. Overall, these results support RNAi-mediated silencing of Foxo3 as an effective strategy to enhance the therapeutic antitumor effect of HER-2/neu DNA vaccines against p185neu-positive tumors.

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Acknowledgements

This study was supported by grants from the Taichung Veterans General Hospital and National Chung Hsing University (TCVGH-NCHU-997602) Taichung, Taiwan and in part by the Ministry of Education, Taiwan, ROC under the ATU plan.

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Author notes

  1. S-T Wang and C-C Chang: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Biomedical Sciences, College of Life Science, National Chung Hsing University, Taichung, Taiwan

    S-T Wang, C-C Chang, Y-T Peng, D-Y Chen, J-L Lan & C-C Lin

  2. Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan

    C-C Chang

  3. Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    M-C Yen & C-F Tu

  4. Yeastern Biotech Co, Taipei County, Taiwan

    C-L Chu

  5. Division of Allergy, Department of Internal Medicine, Immunology and Rheumatology, Taichung Veterans General Hospital, Taichung, Taiwan

    D-Y Chen, J-L Lan & C-C Lin

  6. Department of Internal Medicine, Taichung Veterans General Hospital, Taichung, Taiwan

    J-L Lan

  7. Department of Medical Research and Education, Taichung Veterans General Hospital, Taichung, Taiwan

    C-C Lin

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  1. S-T Wang
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Wang, ST., Chang, CC., Yen, MC. et al. RNA interference-mediated silencing of Foxo3 in antigen-presenting cells as a strategy for the enhancement of DNA vaccine potency. Gene Ther 18, 372–383 (2011). https://doi.org/10.1038/gt.2010.146

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