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Nonviral gene delivery to human breast cancer cells by targeted Ad5 penton proteins

Gene Therapy volume 8, pages 1753–1761 (2001)Cite this article

Abstract

The capsid proteins of adenovirus serotype 5 (Ad5) are key to the virus’ highly efficient cell binding and entry mechanism. In particular, the penton base plays a significant role in both viral internalization and endosome penetration. We have produced an adenovirus penton fusion protein (HerPBK10) containing moieties for DNA transport and targeted delivery to breast cancer cells. HerPBK10 binds DNA through a polylysine appendage, while the EGF-like domain of the heregulin-α1 isoform is used as the targeting ligand. This ligand binds with high affinity to HER2/3 or HER2/4 heterodimers, which are overexpressed on certain aggressive breast cancers. In addition, this ligand is rapidly internalized after binding, thus adding to the utility of heregulin for targeting. HerPBK10 binds MDA-MB-453 breast cancer cells in a receptor-specific manner, and mediates the entry of a reporter plasmid in MDA-MB-453 cells in culture. Delivery can be competed by excess heregulin peptide, thus confirming receptor specificity. Importantly, the penton segment appears to contribute significantly to enhanced delivery. Complexes containing HerPBK10 and DNA have been optimized to provide targeted gene delivery to breast cancer cells in vitro. We demonstrate that delivery can be accomplished in the presence of serum, thus suggesting a potential use for in vivo delivery.

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Acknowledgements

We are grateful to the following people for ongoing discussions and support: Xinhua Chen, Gene Chung, Yasuo Hamamori, John Hwang, Tatsuya Iso, Kimi Kong, Vivian Leung, Jochen Muller-Ehmsen, Alex Oxyzoglou, Coralie Poizat, Tsuyoshi Sakoda, Vittorio Sartorelli, Terry Saluna, David Tinsley and Hung-Yi Wu. Grants to LHK from the Susan G Komen Breast Cancer foundation, the Department of Defense, and the National Institutes of Health (CA59318-07) and a fellowship to LKM-K from the National Institutes of Health (HL09638-02) supported this work.

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Authors and Affiliations

  1. Institute for Genetic Medicine and Department of Biochemistry and Molecular Biology, University of Southern California Keck School of Medicine, Los Angeles, CA, USA

    LK Medina-Kauwe, M Maguire, N Kasahara & L Kedes

  2. Department of Pathology, University of Southern California Keck School of Medicine, Los Angeles, CA, USA

    N Kasahara

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  1. LK Medina-Kauwe
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  2. M Maguire
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  4. L Kedes
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Medina-Kauwe, L., Maguire, M., Kasahara, N. et al. Nonviral gene delivery to human breast cancer cells by targeted Ad5 penton proteins. Gene Ther 8, 1753–1761 (2001). https://doi.org/10.1038/sj.gt.3301583

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