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Adenovirus-Mediated Transfer of the Herpes Simplex Virus Thymidine Kinase Gene Inhibits Vascular Smooth Muscle Cell Proliferation and Neointima Formation Following Balloon Angioplasty of the Rat Carotid Artery

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Abstract

Background

Vascular smooth muscle cell (VSMC) proliferation following arterial injury plays a critical role in a variety of vascular proliferative disorders, including atherosclerosis and restenosis after balloon angioplasty. In this study, we tested the hypothesis that localized arterial infection at the time of balloon angioplasty with an adenovirus (ADV-tk) encoding the herpes simplex virus thymidine kinase gene (HSV-tk), followed by systemic ganciclovir administration, can inhibit VSMC proliferation and neointima formation in a well-characterized model of arterial injury and restenosis.

Materials and Methods

The left carotid arteries of 31 male Sprague-Dawley rats were subjected to balloon angioplasty and immediately infected with 2 × 109 pfu of either ADV-tk or a control adenovirus that does not encode a recombinant protein (ADV-ΔE1). Twenty-four hours after injury, animals from each experimental group were randomized to receive a course of systemic ganciclovir (ADV-tk/+GC, ADV-ΔE1/+GC) or saline (ADV-tk/-GC, ADV-ΔE1/-GC). VSMC DNA synthesis was measured by 5’-bromode-oxyuridine (BrdU) incorporation 2–4 days after balloon injury. The extent of restenosis, expressed as the neointima to media (I/M) area ratio was determined by digital planimetry 20 days after balloon injury in each of the four treatment groups. Immunohistochemistry using a mAb to von Willebrand factor (vWF) was used to determine the effects of ADV-tk infection and ganciclovir treatment on re-endothelialization of the carotid arteries 20 days following balloon angioplasty.

Results

Forty-one percent of the medial VSMCs in the ADV-tk/-GC arteries were labeled with BrdU 4 days after balloon injury. In contrast, ADV-tk infected animals that were treated with systemic ganciclovir (ADV-tk/+GC) displayed a 40% reduction in BrdU-staining medial VSMCs (p < 0.03). I/M area ratios of the three control groups were 1.17 ± 0.18 (ADV-tk/-GC, n = 5), 1.15 ± 0.10 (ADV-ΔE1/+GC, n = 6), and 0.91 ± 0.08 (ADV-ΔE1/-GC, n = 6). These differences were not statistically significant (p > 0.05). In contrast, the ADV-tk/+GC animals (n = 6) displayed an I/M area ratio of 0.49 ±0.13 which was significantly lower than that seen in each of the three control groups (p < 0.02). None of the treated animals showed evidence of significant organ toxicity at autopsy. A regenerated endothelium was observed in the ADV-tk/ +GC animals 20 days after balloon injury.

Conclusions

Localized arterial infection with ADV-tk at the time of balloon angioplasty followed by systemic ganciclovir therapy reduces VSMC proliferation and neointimal expansion in the rat carotid artery injury model. Moreover, combined treatment with ADV-tk and systemic ganciclovir does not result in systemic toxicity and appears to selectively eliminate proliferating VSMCs, while preserving the capacity of the injured arterial segments to re-endothelialize within 3 weeks of injury. Taken together, these results support the feasibility of using this gene therapy approach for the treatment of human vascular proliferative disorders.

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Acknowledgments

The authors would like to thank Lingling Xu and Hong San, MD, for their technical assistance. This work was supported in part by a grant from Vical Inc., La Jolla, CA.

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Authors and Affiliations

  1. Department of Medicine, University of Chicago, 5841 S. Maryland Avenue, Chicago, Illinois, 60637, USA

    Mark W. Chang, Min Min Lu & Jeffrey M. Leiden

  2. Department of Pathology, University of Chicago, Chicago, Illinois, USA

    Jeffrey M. Leiden

  3. Department of Medicine, University of Michigan, Ann Arbor, Michigan, USA

    Takeshi Ohno & Gary J. Nabel

  4. Department of Pathology, University of Michigan, Ann Arbor, Michigan, USA

    David Gordon

  5. Howard Hughes Medical Institute, University of Michigan, Ann Arbor, Michigan, USA

    Gary J. Nabel & Elizabeth G. Nabel

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Chang, M.W., Ohno, T., Gordon, D. et al. Adenovirus-Mediated Transfer of the Herpes Simplex Virus Thymidine Kinase Gene Inhibits Vascular Smooth Muscle Cell Proliferation and Neointima Formation Following Balloon Angioplasty of the Rat Carotid Artery. Mol Med 1, 172–181 (1995). https://doi.org/10.1007/BF03401565

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