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In vitro and in vivo analysis of expression cassettes designed for vascular gene transfer

Gene Therapy volume 15, pages 340–346 (2008)Cite this article

Abstract

Increasing the level and duration of transgene expression and restricting expression to vascular cells are important goals for clinically useful gene therapy vectors. We evaluated several promoters, enhancers and introns in endothelial, smooth muscle and liver cells in tissue culture and in vivo, comparing local delivery to the carotid artery with intravenous delivery to the liver. A 1800-bp fragment of the oxidized LDL receptor (LOX-1) promoter showed highest in vivo activity in the carotid artery, achieving 39% the activity of the reference cytomegalovirus promoter, with 188-fold greater specificity for carotid artery over liver. An enhancer from the Tie2 gene in combination with the intracellular adhesion molecule-2 promoter improved endothelial specificity of plasmid vectors, increased the expression from adenoviral vectors in cultured endothelial cells and doubled the specificity for carotid artery over liver in vivo. Adding a short intron to expression cassettes increased expression in both endothelial and smooth muscle cells in vitro; however, the eNOS enhancer failed to consistently increase the expression or endothelial specificity of the vector. In conclusion, elements from the LOX-1 promoter and Tie2 enhancer together with an intron can be used to improve vectors for vascular gene transfer.

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Acknowledgements

We thank Ray Bush (University of Bristol) for help with animal work; Jill Tarlton and Melanie Roberts (University of Bristol) for help with virus production and technical assistance; James Uney (University of Bristol) for plasmid pXCX-CMV;34 Mike Antoniou (Kings College, London) for the provision of Cβ88 plasmid, containing the Δ89 truncated β-globin 2nd intron23 (intron B); Alessandra Tessitore (University of L'Aquila) for plasmid LC120 containing full ICAM-1 promoter; Anja Ehrhardt (Stanford University) for plasmid pHM5/hFIX.30 This work was performed at the Bristol Heart Institute, the University of Bristol. SJW was supported by British Heart Foundation intermediate fellowship (FS/03/096/16318).

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Author notes

  1. E D Papadakis

    Present address: 3Current address: Kingswood Court, London, UK,

Authors and Affiliations

  1. Bristol Heart Institute, University of Bristol, Bristol, UK

    S J White, E D Papadakis, C A Rogers, J L Johnson & A C Newby

  2. Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands

    E A L Biessen

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  1. S J White
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Correspondence to S J White.

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White, S., Papadakis, E., Rogers, C. et al. In vitro and in vivo analysis of expression cassettes designed for vascular gene transfer. Gene Ther 15, 340–346 (2008). https://doi.org/10.1038/sj.gt.3303058

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