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Alternatively spliced vascular endothelial growth factor receptor-2 is an essential endogenous inhibitor of lymphatic vessel growth

Abstract

Disruption of the precise balance of positive and negative molecular regulators of blood and lymphatic vessel growth can lead to myriad diseases. Although dozens of natural inhibitors of hemangiogenesis have been identified, an endogenous selective inhibitor of lymphatic vessel growth has not to our knowledge been previously described. We report the existence of a splice variant of the gene encoding vascular endothelial growth factor receptor-2 (Vegfr-2) that encodes a secreted form of the protein, designated soluble Vegfr-2 (sVegfr-2), that inhibits developmental and reparative lymphangiogenesis by blocking Vegf-c function. Tissue-specific loss of sVegfr-2 in mice induced, at birth, spontaneous lymphatic invasion of the normally alymphatic cornea and hyperplasia of skin lymphatics without affecting blood vasculature. Administration of sVegfr-2 inhibited lymphangiogenesis but not hemangiogenesis induced by corneal suture injury or transplantation, enhanced corneal allograft survival and suppressed lymphangioma cellular proliferation. Naturally occurring sVegfr-2 thus acts as a molecular uncoupler of blood and lymphatic vessels; modulation of sVegfr-2 might have therapeutic effects in treating lymphatic vascular malformations, transplantation rejection and, potentially, tumor lymphangiogenesis and lymphedema (pages 993–994)

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Figure 1: Loss of endogenous sVegfr-2, which antagonizes Vegf-c, leads to spontaneous corneal lymphangiogenesis.
Figure 2: sVegfr-2 inhibits reparative corneal lymphangiogenesis and rejection of corneal allografts.
Figure 3: Endogenous Vegf-c and sVegfr-2 selectively modulate corneal lymphangiogenesis.
Figure 4: Loss of sVegfr2 in the skin induces lymphatic hyperplasia.
Figure 5: sVegfr-2 is produced by BECs and skin epithelium and circulates in plasma.
Figure 6: sVEGFR-2 exists in humans and inhibits human lymphangioma cell proliferation.

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Acknowledgements

We thank R. Ashery-Padan (Tel Aviv University), P. Gruss (Max Planck Institute) and D.C. Beebe (Washington University) for LeCre mice, R.K. Nordeen (University of Colorado) for Cre plasmid, K. Miyazono (University of Tokyo) for pVegf-c and J.L. Arbiser (Emory University) and C.D. Kontos (Duke University) for endothelial cell lines; R. King, L. Xu, M. McConnell, K. Emerson, A. Blanford, M. Baker, S. Furlow, M. LaFalce and C. Long for technical assistance; and R. Mohan, F. Cambi, S. Bondada, M. Detmar, M.W. Fannon, T.V. Getchell, R.K. Jain, T.S. Khurana, B.J. Raisler, J.E. Springer, P.A. Pearson, C.W. Vander Kooi, J.G. Woodward, A.M. Rao, G.S. Rao, K. Ambati and L. Garcia for valuable discussions. This work was supported by US National Institutes of Health and National Eye Institute grants EY015422, EY018350 and EY018836 to J.A. and EY017182 and EY017950 to B.K.A.; a Research to Prevent Blindness Lew R. Wassermann Merit Award (J.A.); Physician Scientist Awards (J.A., B.K.A.); a Medical Student Fellowship (R.J.C.A.); a departmental unrestricted grant (J.A.); a University of Kentucky University Research Professorship (J.A.); Fight for Sight (R.J.C.A.); Japan Society for the Promotion of Science for Young Scientists (A.T.); a VA Merit Award (B.K.A.); and the US Department of Defense (B.K.A.). J.A. is also supported by the Doris Duke Distinguished Clinical Scientist Award and the Burroughs Wellcome Fund Clinical Scientist Award in Translational Research and is the Dr. E. Vernon Smith and Eloise C. Smith Macular Degeneration Endowed Chair.

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R.J.C.A., T.H., W.G.C., M.E.K., S.D., A.T., J.Z.B., K.Y., H.K., M.G.G. and S.C. designed and conducted experiments. R.A.B., J.W., H.A.W. and J.S.A. provided reagents and, with J.C., S.Y., S.A., N.M., M.L.P., M.H., T.U. and B.K.A., participated in planning experiments. J.A. conceived of and directed the project. R.J.C.A. and J.A. wrote the manuscript. All authors had the opportunity to discuss the results and comment on the manuscript.

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Correspondence to Jayakrishna Ambati.

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J.A. and R.J.C.A. are named as inventors in a patent application to the US Patent and Trademark Office filed by the University of Kentucky surrounding the intellectual property described in this paper (gene and protein sequences and therapeutic application of augmenting or diminishing soluble vascular endothelial growth factor receptor-2 amounts). As employees of the University of Kentucky, they are covered under the distribution of royalties policies of the institution.

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Albuquerque, R., Hayashi, T., Cho, W. et al. Alternatively spliced vascular endothelial growth factor receptor-2 is an essential endogenous inhibitor of lymphatic vessel growth. Nat Med 15, 1023–1030 (2009). https://doi.org/10.1038/nm.2018

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