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Tissue-specific expression of an anti-proliferative hybrid transgene from the human smooth muscle α-actin promoter suppresses smooth muscle cell proliferation and neointima formation

Gene Therapy volume 8, pages 1847–1854 (2001)Cite this article

Abstract

The retinoblastoma protein (Rb), a key regulator of cell cycle progression, can bind the transcription factor E2F converting it from a positive transcriptional factor capable of driving cells into S phase into a negative complex which arrests cells in G1. We have created a potent transcriptional repressor of E2F-dependent transcription by fusing the C-terminal fragment of Rb (p56) to the DNA and DP1-binding domains of E2F. Because the expression of E2F/56 fusion protein from a constitutive promoter was incompatible with virus growth, adenovirus constructs were prepared where transgenes were expressed from a fragment of the smooth muscle α-actin (SMA) promoter. Immunoblot and β-galactosidase staining demonstrated smooth muscle-specific expression of this transcriptional element in vitro. The SMA-p56 and SMA-E2F/p56 adenoviral constructs also induced G0/G1 cell cycle arrest specifically in smooth muscle cells. Following administration to rat tissues, the SMA-β-galactosidase construct exhibited expression in balloon-injured carotid arteries, but not in liver, bladder or skeletal muscle. Local delivery of the SMA-E2F/p56 adenoviral construct to balloon-injured carotid arteries inhibited intimal hyperplasia. Our results demonstrate that local delivery of the SMA-E2F/p56 adenoviral construct can limit intimal hyperplasia in balloon-injured vessels, while avoiding toxicity that could occur from the dissemination and expression of the viral transgene.

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Authors and Affiliations

  1. Canji, Inc., San Diego, CA, USA

    KN Wills, JB Avanzini & D Antelman

  2. Division of Cardiovascular Research, Department of Medicine, St Elizabeth's Medical Center, Boston, MA, USA

    T Mano, T Nguyen, RC Smith & K Walsh

  3. Genzyme, Framingham, MA, USA

    RJ Gregory

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  1. KN Wills
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  2. T Mano
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  3. JB Avanzini
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  4. T Nguyen
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  5. D Antelman
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  7. RC Smith
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  8. K Walsh
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Wills, K., Mano, T., Avanzini, J. et al. Tissue-specific expression of an anti-proliferative hybrid transgene from the human smooth muscle α-actin promoter suppresses smooth muscle cell proliferation and neointima formation. Gene Ther 8, 1847–1854 (2001). https://doi.org/10.1038/sj.gt.3301603

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  • DOI: https://doi.org/10.1038/sj.gt.3301603

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