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Use of protamine to augment adenovirus-mediated cancer gene therapy

Gene Therapy volume 6, pages 1600–1610 (1999)Cite this article

Abstract

Improving the therapeutic potential of adenoviral (Ad) suicide gene therapy has become an area of intense investigation since the inception of gene therapy strategies for cancer treatment. Poor efficiency of gene transfer to target tissues has become one of the most important limitations to Ad-based gene therapy. Since polycations have been shown to enhance adenovirus-mediated gene transfer in epithelial cells both in vitro and in vivo, we hypothesized that polycations could augment treatment efficacy in animals with established tumor. To address this hypothesis, protamine sulfate, a polycation already safely administered in humans, was complexed with a recombinant Ad (E1E3-deleted) vector containing the herpes simplex 1 thymidine kinase (HSVtk) suicide gene to treat cancer cell lines in vitro and in animals bearing intraperitoneal tumor. In the presence of 5 μg/ml protamine, the efficiency of gene transfer to a number of cancer cell lines normally resistant to adenovirus was significantly enhanced. Protamine’s effect in vitro was found to be inversely proportional to the level of expression of the high affinity Ad binding site, coxsackievirus and adenovirus receptor (CAR), on the sur- face of the various cell lines tested. Ad.tk infected tumor cells were rendered 2.5- to three-fold more sensitive to 20 μM ganciclovir (GCV) in the presence of protamine. Protamine also augmented the in vivo transfer efficiency of the marker gene, LacZ (contained in an Ad vector), on the surface of tumors derived from an intraperitoneal mouse model. Quantitative imaging revealed 50% tumor surface transduced with LacZ when treatment was performed in the presence of 50 μg/ml protamine compared with 12% tumor surface in controls. However, experiments performed utilizing intraperitoneal administration of Ad.tk/GCV in the presence or absence of 50 μg/ml protamine demonstrated no significantly improved median survival in mice bearing established intraperitoneal tumors. Similarly, in Fischer rats bearing intrapleural tumor, no improvement in anti-tumor response was observed when Ad treatment was performed intrapleurally in the presence of protamine. Thus, although protamine induced an enhancement of Ad-mediated gene transfer in vitro and in vivo, its use as an adjunct to intracavitary Ad-based cancer gene therapy in vivo appears to be limited.

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Acknowledgements

This research was supported by a grant from the National Cancer Institute (NCI PO1 66726). Partial support provided by the Samuel H Lunenfeld Charitable Foundation and the Benjamin Shein Foundation for Humanity. The authors gratefully acknowledge Selim M Arcasoy, MD and Jeffrey M Bergelson MD for providing constructive commentary on the data presented in this manuscript. We would also like to thank Dr Prem Seth for providing the recombinant adenovirus containing the cytosine deaminase expression cassette (Ad.CMVcd).

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Authors and Affiliations

  1. Department of Surgery, Thoracic Oncology Research Laboratory, Philadelphia, PA, USA

    M Lanuti, S D Force, M Y Chang, K Amin & L R Kaiser

  2. Department of Medicine, Pulmonary/Critical Care Division, Philadelphia, PA, USA

    C El Kouri & S M Albelda

  3. Department of Pharmacology, University of Pennsylvania Medical Center, Philadelphia, PA, USA

    K Xu & I A Blair

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  1. M Lanuti
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Lanuti, M., Kouri, C., Force, S. et al. Use of protamine to augment adenovirus-mediated cancer gene therapy. Gene Ther 6, 1600–1610 (1999). https://doi.org/10.1038/sj.gt.3300987

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