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: Liya Pi, Anitha K Shenoy, Jianwen Liu, Seungbum Kim, Nikole Nelson, Huiming Xia, William W Hauswirth, Bryon E Petersen, Gregory S Schultz, Edward W Scott
• Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology (FASEB J) Vol. 26 Issue 8 Pg. 3365-79 (Aug 2012) ISSN: 1530-6860 [Electronic] United States
• PMID: 22611085 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• CCN2 protein, mouse
• Membrane Proteins
• Slit3 protein, mouse
• Connective Tissue Growth Factor

Topics

• Animals
• Carcinoma, Lewis Lung (chemically induced)
• Connective Tissue Growth Factor (physiology)
• Cystine Knot Motifs (drug effects, genetics)
• Human Umbilical Vein Endothelial Cells
• Humans
• Membrane Proteins (physiology)
• Mice
• Neovascularization, Physiologic (drug effects, physiology)
• Retinal Vessels (growth & development)
• Two-Hybrid System Techniques