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Re-activation of a dormant tumor suppressor gene maspin by designed transcription factors

Oncogene volume 26, pages 2791–2798 (2007)Cite this article

Abstract

The controlled and specific re-activation of endogenous tumor suppressors in cancer cells represents an important therapeutic strategy to block tumor growth and subsequent progression. Other than ectopic delivery of tumor suppressor-encoded cDNA, there are no therapeutic tools able to specifically re-activate tumor suppressor genes that are silenced in tumor cells. Herein, we describe a novel approach to specifically regulate dormant tumor suppressors in aggressive cancer cells. We have targeted the Mammary Serine Protease Inhibitor (maspin) (SERPINB5) tumor suppressor, which is silenced by transcriptional and aberrant promoter methylation in aggressive epithelial tumors. Maspin is a multifaceted protein, regulating tumor cell homeostasis through inhibition of cell growth, motility and invasion. We have constructed artificial transcription factors (ATFs) made of six zinc-finger (ZF) domains targeted against 18-base pair (bp) unique sequences in the maspin promoter. The ZFs were linked to the activator domain VP64 and delivered in breast tumor cells. We found that the designed ATFs specifically interact with their cognate targets in vitro with high affinity and selectivity. One ATF was able to re-activate maspin in cell lines that comprise a maspin promoter silenced by epigenetic mechanisms. Consistently, we found that this ATF was a powerful inducer of apoptosis and was able to knock down tumor cell invasion in vitro. Moreover, this ATF was able to suppress MDA-MB-231 growth in a xenograft breast cancer model in nude mice. Our work suggests that ATFs could be used in cancer therapeutics as novel molecular switches to re-activate dormant tumor suppressors.

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Acknowledgements

We thank Dr S Earp for the critical reading of the paper. We thank Dr MJ Hendrix for sending the maspin-luciferase promoter constructs and Dr M Tschan for sending a p53-expression plasmid. This work was supported by an American Lung Association and LUNGevity Foundation (LD-17098-N), a V-Foundation Award and a DoD idea award (BC051475) to PB.

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Authors and Affiliations

  1. Department of Pharmacology and the Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    A Beltran, S Parikh, Y Liu, B D Cuevas, G L Johnson & P Blancafort

  2. Department of Pharmacology and Toxicology, Arizona Cancer Center and College of Pharmacy, University of Arizona, Tucson, AZ, USA

    B W Futscher

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  1. A Beltran
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  2. S Parikh
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  3. Y Liu
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  4. B D Cuevas
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  5. G L Johnson
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  6. B W Futscher
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  7. P Blancafort
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Correspondence to P Blancafort.

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Supplementary Information accompanies the paper on Oncogene website (http://www.nature.com/onc).

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Beltran, A., Parikh, S., Liu, Y. et al. Re-activation of a dormant tumor suppressor gene maspin by designed transcription factors. Oncogene 26, 2791–2798 (2007). https://doi.org/10.1038/sj.onc.1210072

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