title: Dual suppression of hemangiogenesis and lymphangiogenesis by splice-shifting morpholinos targeting vascular endothelial growth factor receptor 2 (KDR).
abstract: The KDR gene, which participates in angiogenesis and lymphangiogenesis, produces two functionally distinct protein products, membrane-bound KDR (mbKDR) and its isoform, soluble KDR (sKDR). Since sKDR does not have a tyrosine kinase domain and does not dimerize, it is principally an antagonist of lymphangiogenesis by sequestering VEGF-C. Alternative polyadenylation of exon 30 or intron 13 leads to the production of mbKDR or sKDR, respectively, yet the regulatory mechanisms are unknown. Here we show that an antisense morpholino oligomer directed against the exon 13-intron 13 junction increases sKDR (suppressing lymphangiogenesis) and decreases mbKDR (inhibiting hemangiogenesis). The latent polyadenylation site in intron 13 of KDR is activated by blocking the upstream 5' splicing site with an antisense morpholino oligomer. Intravitreal morpholino injection suppressed laser choroidal neovascularization while increasing sKDR. In the mouse cornea, subconjunctival injection of the morpholino-inhibited corneal angiogenesis and lymphangiogenesis, and suppressed graft rejection after transplantation. Thus, this morpholino can be used for concurrent suppression of hemangiogenesis and lymphangiogenesis. This study offers new insight into the mechanisms and potential therapeutic modulation of alternative polyadenylation.
authors.affiliation: Moran Eye Center, University of Utah, Salt Lake City, UT 84132, USA.
authors.forename: Hironori
authors.initials: H
authors.lastname: Uehara
authors.forename: YangKyung
authors.initials: Y
authors.lastname: Cho
authors.forename: Jackie
authors.initials: J
authors.lastname: Simonis
authors.forename: Judd
authors.initials: J
authors.lastname: Cahoon
authors.forename: Bonnie
authors.initials: B
authors.lastname: Archer
authors.forename: Ling
authors.initials: L
authors.lastname: Luo
authors.forename: Subrata K
authors.initials: SK
authors.lastname: Das
authors.forename: Nirbhai
authors.initials: N
authors.lastname: Singh
authors.forename: Jayakrishna
authors.initials: J
authors.lastname: Ambati
authors.forename: Balamurali K
authors.initials: BK
authors.lastname: Ambati
mesh_terms: D000818:Animals; D001483:Base Sequence; D016039:Corneal Transplantation; D017931:DNA Primers; D005091:Exons; D006801:Humans; D020287:In Situ Nick-End Labeling; D042583:Lymphangiogenesis / Q000235:genetics*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D060172:Morpholinos / Q000235:genetics*; D018919:Neovascularization, Physiologic / Q000235:genetics*; D012326:RNA Splicing; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics*
chemical_list: D017931:DNA Primers; D060172:Morpholinos; D040301:Vascular Endothelial Growth Factor Receptor-2