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Induction of angiogenesis in a mouse model using engineered transcription factors

Nature Medicine volume 8, pages 1427–1432 (2002)Cite this article

Abstract

The relationship between the structure of zinc-finger protein (ZFP) transcription factors and DNA sequence binding specificity has been extensively studied1. Advances in this field have made it possible to design ZFPs de novo that will bind to specific targeted DNA sequences2,3,4,5,6,7,8,9,10. It has been proposed that such designed ZFPs may eventually be useful in gene therapy6,7,10. A principal advantage of this approach is that activation of an endogenous gene ensures expression of the natural array of splice variants2. Preliminary studies in tissue culture have validated the feasibility of this approach2,3,4. The studies reported here were intended to test whether engineered transcription factors are effective in a whole-organism model. ZFPs were designed to regulate the endogenous gene encoding vascular endothelial growth factor-A (Vegfa). Expression of these new ZFPs in vivo led to induced expression of the protein VEGF-A, stimulation of angiogenesis and acceleration of experimental wound healing. In addition, the neovasculature resulting from ZFP-induced expression of Vegfa was not hyperpermeable as was that produced by expression of murine Vegfa164 cDNA. These data establish, for the first time, that specifically designed transcription factors can regulate an endogenous gene in vivo and evoke a potentially therapeutic biophysiologic effect.

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Figure 1: Targeting, design and in vitro analysis of zinc-finger proteins designed for the mouse Vegfa locus.
Figure 2: ZFP-mediated Vegfa activation and induction of angiogenesis in vivo.
Figure 3: ZFP-mediated Vegfa activation accelerates wound healing.

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Acknowledgements

The authors thank A. Wolffe whose essential scientific input and discussions made this work possible; C. Pabo for scientific input and helpful comments on the manuscript; E. Lanphier and D. Bobo for supporting the ZFP concept; and A. Vincent for technical assistance. This work was funded by an American Heart Association award (AHA no. 97-30083N), by US National Institutes of Health grant RO1-HL64001 and by an unrestricted gift from Edwards Lifesciences (all to F.J.G.).

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  1. Edward J. Rebar and Yan Huang: These authors contributed equally to this work.

Authors and Affiliations

  1. Sangamo Biosciences, Richmond, California, USA

    Edward J. Rebar, Bingliang Chen, Lei Xu, Yuxin Liang, Andrew C. Jamieson, Lei Zhang, S Kaye Spratt, Casey C. Case & Alan Wolffe

  2. Dept. of Medicine, Yale University School of Medicine, New Haven, Connecticut, USA

    Yan Huang, Reed Hickey, Anjali K. Nath, David Meoli, Sameer Nath & Frank J. Giordano

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  1. Edward J. Rebar
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Correspondence to Frank J. Giordano.

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Competing interests

E.J.R., B.C., L.X., Y.L., A.C.J., L.Z., K.S., C.C.C. and A.W. have potential competing interests in that they are employees of Sangamo Biosciences, which could benefit from the publication of this data.

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Rebar, E., Huang, Y., Hickey, R. et al. Induction of angiogenesis in a mouse model using engineered transcription factors. Nat Med 8, 1427–1432 (2002). https://doi.org/10.1038/nm1202-795

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