Abstract
The genes that regulate the formation of blood vessels in adult tissues represent promising therapeutic targets because angiogenesis plays a role in many diseases, including cancer. We wished to develop a mouse model allowing characterization of gene function in adult angiogenic vasculature while minimizing effects on embryonic vasculature or adult quiescent vasculature. Here we describe a transgenic mouse model that allows expression of proteins in the endothelial cells of newly forming blood vessels in the adult using a selective retroviral gene delivery system. We generated transgenic mouse lines that express the TVA receptor for the RCAS avian-specific retrovirus from Flk1 gene regulatory elements that drive expression in proliferating endothelial cells. Several of these Flk1-TVA lines expressed TVA mRNA in the embryonic vasculature and TVA protein in new blood vessels growing into subcutaneous extracellular matrix implants in adult mice. In a Flk1-TVA line that was crossed with the MMTV-PyMT transgenic mammary tumor model, tumor endothelial cells also expressed the TVA protein. Furthermore, endothelial cells in extracellular matrix implants and the tumors of Flk1-TVA mice were susceptible to RCAS infection, as determined by expression of green fluorescent protein encoded by the virus. The Flk1-TVA mouse model in conjunction with the RCAS gene delivery system will be useful to study molecular mechanisms underlying adult forms of angiogenesis.
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Acknowledgments
The authors thank H. E. Varmus for the TVA cDNA, S. Hughes for the RCAS and pCla12 plasmids, M. Haden and L. Soroceanu for subcloning the TVA cDNA into several vectors, J. A. Seddon for preparing the Flk1-TVA DNA for injection, L. Wang for performing the oocyte microinjection experiments, V. Munoz for preliminary tumor infection trials and and S. Codeluppi for sharing unpublished information on RCAS–EGFP infection of chicken embryonic retina. This work was supported by DOD BCRP grant DAMD17-02-1-0314 and NIH CA102583.
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Vervoort, V.S., Lu, M., Valencia, F. et al. A novel Flk1-TVA transgenic mouse model for gene delivery to angiogenic vasculature. Transgenic Res 17, 403–415 (2008). https://doi.org/10.1007/s11248-007-9156-1
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DOI: https://doi.org/10.1007/s11248-007-9156-1