Vasohibin as an endothelium-derived negative feedback regulator of angiogenesis
Kazuhide Watanabe, … , Hikaru Sonoda, Yasufumi Sato
Kazuhide Watanabe, … , Hikaru Sonoda, Yasufumi Sato
Published October 1, 2004
Citation Information: J Clin Invest. 2004;114(7):898-907. https://doi.org/10.1172/JCI21152.
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Article Angiogenesis

Vasohibin as an endothelium-derived negative feedback regulator of angiogenesis

Abstract

Negative feedback is a crucial physiological regulatory mechanism, but no such regulator of angiogenesis has been established. Here we report a novel angiogenesis inhibitor that is induced in endothelial cells (ECs) by angiogenic factors and inhibits angiogenesis in an autocrine manner. We have performed cDNA microarray analysis to survey VEGF-inducible genes in human ECs. We characterized one such gene, KIAA1036, whose function had been uncharacterized. The recombinant protein inhibited migration, proliferation, and network formation by ECs as well as angiogenesis in vivo. This inhibitory effect was selective to ECs, as the protein did not affect the migration of smooth muscle cells or fibroblasts. Specific elimination of the expression of KIAA1036 in ECs restored their responsiveness to a higher concentration of VEGF. The expression of KIAA1036 was selective to ECs, and hypoxia or TNF-α abrogated its inducible expression. As this molecule is preferentially expressed in ECs, we designated it “vasohibin.” Transfection of Lewis lung carcinoma cells with the vasohibin gene did not affect the proliferation of cancer cells in vitro, but did inhibit tumor growth and tumor angiogenesis in vivo. We propose vasohibin to be an endothelium-derived negative feedback regulator of angiogenesis.

Authors

Kazuhide Watanabe, Yasuhiro Hasegawa, Hiroshi Yamashita, Kazue Shimizu, Yuanying Ding, Mayumi Abe, Hideki Ohta, Keiichi Imagawa, Kanji Hojo, Hideo Maki, Hikaru Sonoda, Yasufumi Sato

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Modulation of vasohibin expression and the effect of vasohibin on VEGF-s...
Modulation of vasohibin expression and the effect of vasohibin on VEGF-stimulated signaling in HUVECs. (A) Induction of vasohibin. HUVECs were stimulated with VEGF (1 nM), PlGF (1 nM), FGF-2 (2 nM), HGF (1 nM), TNF-α (1 nM), or 10% serum for 24 hours. Thereafter, total RNA was obtained and Northern blotting for vasohibin was performed. (B) Effect of TNF-α on the induction of vasohibin by VEGF. HUVECs were stimulated with VEGF (1 nM) and/or TNF-α (1 nM). Thereafter, Northern blotting and Western blotting for vasohibin were performed. (C) Effect of hypoxia on the induction of vasohibin by VEGF. HUVECs were stimulated with VEGF (1 nM) under normoxic (N) or hypoxic (H) conditions. Upper panel: Total RNA was obtained and real-time RT-PCR of vasohibin was performed. Values are expressed as mean ± SD of 4 samples. **P < 0.01. Lower panel: Cell extract was obtained and Western blotting for vasohibin was performed. (D) Effect of vasohibin on VEGF-mediated KDR tyrosine phosphorylation or ERK1/2 activation of HUVECs. HUVECs were infected with AdLacZ or AdKIAA at an MOI of 100, and then stimulated with VEGF (10 ng/ml). VEGF-mediated KDR tyrosine phosphorylation or ERK1/2 activation was analyzed. Results shown in lower panel indicate that AdKIAA increased the synthesis of vasohibin in an MOI-dependent manner. IP, immunoprecipitation; IB, immunoblotting; pKDR, phosphorylated KDR.