VEGF (Vascular Endothelial Growth Factor) Induces NRP1 (Neuropilin-1) Cleavage via ADAMs (a Disintegrin and Metalloproteinase) 9 and 10 to Generate Novel Carboxy-Terminal NRP1 Fragments That Regulate Angiogenic Signaling.

Publisher: Lippincott Williams & Wilkins Country of Publication: United States NLM ID: 9505803 Publication Model: Print Cited Medium: Internet ISSN: 1524-4636 (Electronic) Linking ISSN: 10795642 NLM ISO Abbreviation: Arterioscler Thromb Vasc Biol Subsets: MEDLINE

Publication: 1998- : Baltimore, Md. : Lippincott Williams & Wilkins
Original Publication: Dallas, TX : American Heart Association, c1995-

ADAMTS Proteins/*metabolism ; ADAMTS9 Protein/*metabolism ; Angiogenesis Inducing Agents/*pharmacology ; Human Umbilical Vein Endothelial Cells/*drug effects ; Neovascularization, Physiologic/*drug effects ; Neuropilin-1/*metabolism ; Peptide Fragments/*metabolism ; Vascular Endothelial Growth Factor A/*pharmacology; ADAMTS Proteins/genetics ; ADAMTS9 Protein/genetics ; Animals ; Cell Movement/drug effects ; Cells, Cultured ; Coculture Techniques ; Human Umbilical Vein Endothelial Cells/enzymology ; Humans ; Mice ; Neuropilin-1/genetics ; Phosphorylation ; Protein Binding ; Protein Interaction Domains and Motifs ; Proteolysis ; Signal Transduction/drug effects ; Vascular Endothelial Growth Factor Receptor-2/metabolism

Objective- NRP1(neuropilin-1) acts as a coreceptor for VEGF (vascular endothelial growth factor) with an essential role in angiogenesis. Recent findings suggest that posttranslational proteolytic cleavage of VEGF receptors may be an important mechanism for regulating angiogenesis, but the role of NRP1 proteolysis and the NRP1 species generated by cleavage in endothelial cells is not known. Here, we characterize NRP1 proteolytic cleavage in endothelial cells, determine the mechanism, and investigate the role of NRP1 cleavage in regulation of endothelial cell function. Approach and Results- NRP1 species comprising the carboxy (C)-terminal and transmembrane NRP1 domains but lacking the ligand-binding A and B regions are constitutively expressed in endothelial cells. Generation of these C-terminal domain NRP1 proteins is upregulated by phorbol ester and Ca ²⁺ ionophore, and reduced by pharmacological inhibition of metalloproteinases, by small interfering RNA-mediated knockdown of 2 members of ADAM (a disintegrin and metalloproteinase) family, ADAMs 9 and 10, and by a specific ADAM10 inhibitor. Furthermore, VEGF upregulates expression of these NRP1 species in an ADAM9/10-dependent manner. Transduction of endothelial cells with adenoviral constructs expressing NRP1 C-terminal domain fragments inhibited VEGF-induced phosphorylation of VEGFR2 (VEGF receptor tyrosine kinase)/KDR (kinase domain insert receptor) and decreased VEGF-stimulated endothelial cell motility and angiogenesis in coculture and aortic ring sprouting assays. Conclusions- These findings identify novel NRP1 species in endothelial cells and demonstrate that regulation of NRP1 proteolysis via ADAMs 9 and 10 is a new regulatory pathway able to modulate VEGF angiogenic signaling.

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RG/06/003 United Kingdom BHF_ British Heart Foundation; PG/17/87/33400 United Kingdom BHF_ British Heart Foundation; RG/11/11/29050 United Kingdom BHF_ British Heart Foundation; PG/16/84/32464 United Kingdom BHF_ British Heart Foundation; PG/12/65/29840 United Kingdom BHF_ British Heart Foundation

Keywords: cytokines; endothelial cells; phosphorylation; proteolysis; signal transduction

0 (Angiogenesis Inducing Agents)
0 (NRP1 protein, human)
0 (Peptide Fragments)
0 (VEGFA protein, human)
0 (Vascular Endothelial Growth Factor A)
144713-63-3 (Neuropilin-1)
EC 2.7.10.1 (KDR protein, human)
EC 2.7.10.1 (Vascular Endothelial Growth Factor Receptor-2)
EC 3.4.24.- (ADAMTS Proteins)
EC 3.4.24.- (ADAMTS10 protein, human)
EC 3.4.24.- (ADAMTS9 Protein)
EC 3.4.24.- (ADAMTS9 protein, human)

Date Created: 20180609 Date Completed: 20190722 Latest Revision: 20210510

20231215

PMC6092111

10.1161/ATVBAHA.118.311118

29880492