title: CCN2/CTGF regulates neovessel formation via targeting structurally conserved cystine knot motifs in multiple angiogenic regulators.
abstract: Blood vessels are formed during development and tissue repair through a plethora of modifiers that coordinate efficient vessel assembly in various cellular settings. Here we used the yeast 2-hybrid approach and demonstrated a broad affinity of connective tissue growth factor (CCN2/CTGF) to C-terminal cystine knot motifs present in key angiogenic regulators Slit3, von Willebrand factor, platelet-derived growth factor-B, and VEGF-A. Biochemical characterization and histological analysis showed close association of CCN2/CTGF with these regulators in murine angiogenesis models: normal retinal development, oxygen-induced retinopathy (OIR), and Lewis lung carcinomas. CCN2/CTGF and Slit3 proteins worked in concert to promote in vitro angiogenesis and downstream Cdc42 activation. A fragment corresponding to the first three modules of CCN2/CTGF retained this broad binding ability and gained a dominant-negative function. Intravitreal injection of this mutant caused a significant reduction in vascular obliteration and retinal neovascularization vs. saline injection in the OIR model. Knocking down CCN2/CTGF expression by short-hairpin RNA or ectopic expression of this mutant greatly decreased tumorigenesis and angiogenesis. These results provided mechanistic insight into the angiogenic action of CCN2/CTGF and demonstrated the therapeutic potential of dominant-negative CCN2/CTGF mutants for antiangiogenesis.
authors.affiliation: Program in Stem Cell Biology and Regenerative Medicine, University of Florida, PO Box 100232, Gainesville, FL 32610, USA. lpi@ufl.edu
authors.forename: Liya
authors.initials: L
authors.lastname: Pi
authors.forename: Anitha K
authors.initials: AK
authors.lastname: Shenoy
authors.forename: Jianwen
authors.initials: J
authors.lastname: Liu
authors.forename: Seungbum
authors.initials: S
authors.lastname: Kim
authors.forename: Nikole
authors.initials: N
authors.lastname: Nelson
authors.forename: Huiming
authors.initials: H
authors.lastname: Xia
authors.forename: William W
authors.initials: WW
authors.lastname: Hauswirth
authors.forename: Bryon E
authors.initials: BE
authors.lastname: Petersen
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Schultz
authors.forename: Edward W
authors.initials: EW
authors.lastname: Scott
mesh_terms: D000818:Animals; D018827:Carcinoma, Lewis Lung / Q000139:chemically induced; D055513:Connective Tissue Growth Factor / Q000502:physiology*; D047168:Cystine Knot Motifs / Q000187:drug effects* / Q000235:genetics; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008565:Membrane Proteins / Q000502:physiology; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000502:physiology*; D012171:Retinal Vessels / Q000254:growth & development; D020798:Two-Hybrid System Techniques
chemical_list: C526318:CCN2 protein, mouse; D008565:Membrane Proteins; C432251:Slit3 protein, mouse; D055513:Connective Tissue Growth Factor