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Immunocompetent mouse model of breast cancer for preclinical testing of EphA2-targeted therapy

Cancer Gene Therapy volume 12, pages 46–53 (2005)Cite this article

Abstract

EphA2, a receptor tyrosine kinase, is elevated in many invasive human breast cancers, and the majority of EphA2 remains unphosphorylated. The successful attachment of ligand EphrinA1 present on the surface of adjacent cells to EphA2 initiates EphA2 phosphorylation leading to its turnover. In vivo efficacy of various approaches targeting EphA2 for breast cancer therapy is usually evaluated in nude mice bearing human breast cancer xenografts. In order to establish an immunocompetent mouse model of breast cancer for EphA2-targeted therapies, we evaluated a mouse breast cancer cell line (MT1A2) for EphA2 expression and phosphorylation. Overexpression of EphA2 was observed in MT1A2 cells and the majority of it remained unphosphorylated signifying that EphA2 in MT1A2 cells behaved similar to that of human breast cancer cells. Human adenovirus subtype 5 (HAd5) vectors expressing secretory forms of EphrinA1 were used for in vitro and in vivo targeting of MT1A2-derived EphA2. MT1A2 cells infected with HAd-EphrinA1-Fc (HAd expressing extracellular domain of human EphrinA1 attached to Fc portion of human IgG1 heavy chain) induced EphA2 activation and its turnover. This led to inhibition in MT1A2 cell colony formation in soft agar and cell viability in monolayer culture. In addition, MT1A2 cells-infected with HAd-EphrinA1-Fc failed to form tumors in syngeneic FVB/n mice at least 32 days postinoculation. Moreover, intratumoral inoculation of FVB/n mice-bearing MT1A2-induced tumors with HAd-EphrinA1-Fc slowed the tumor growth and also resulted in the development of vector-specific immune response. These results indicate that FVB/n mice-bearing MT1A2-induced tumors could serve as an immunocompetent model of breast cancer for EphA2-targeted therapeutic strategies.

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Acknowledgements

We thank Jane Stewart, Shaji Abraham, Rebecca Pratt, and Keith Kikawa for their technical advice and Jane Kovach for excellent secretarial assistance. We are also thankful to Dr Mike Kinch, MedImmune, Inc., Gaithersburg, MD for discussions and for providing EphA2-specific reagents, and Dr William Muller, Department of Biology, McMaster University, Hamilton, Ontario, Canada for providing MT1A2 cell line. This work was supported by a Purdue Research Foundation Grant.

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Authors and Affiliations

  1. Laboratory of Gene Therapy, Purdue University, West Lafayette, Indiana, 47907, USA

    Loren W Noblitt, Dinesh S Bangari, Shruti Shukla & Suresh K Mittal

  2. Purdue University Cancer Center, Purdue University, West Lafayette, Indiana, 47907, USA

    Loren W Noblitt, Dinesh S Bangari, Shruti Shukla, Sulma Mohammed & Suresh K Mittal

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  1. Loren W Noblitt
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Correspondence to Suresh K Mittal.

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Noblitt, L., Bangari, D., Shukla, S. et al. Immunocompetent mouse model of breast cancer for preclinical testing of EphA2-targeted therapy. Cancer Gene Ther 12, 46–53 (2005). https://doi.org/10.1038/sj.cgt.7700763

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