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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KIAA1036 inhibits angiogenesis. (A) Preparation of KIAA1036 protein. SDS...
KIAA1036 inhibits angiogenesis. (A) Preparation of KIAA1036 protein. SDS-PAGE/Coomassie brilliant blue staining is shown on the left and Western blotting on the right. Arrows indicate KIAA1036. (B) Effect of KIAA1036 on network formation by ECs. HUVECs were plated on Matrigel in the absence or presence of KIAA1036 (10 nM). (C) Downregulation of KIAA1036 during network formation on Matrigel. HUVECs were plated on Matrigel, and after the indicated period of incubation, total RNA was obtained and real-time RT-PCR was performed. Values are expressed as mean ± SD of 3 samples. (D) Matrigel implantation analysis was performed as described in Methods. The vessel number per low-power field in 3 different fields was counted for each sample. Values are expressed as mean ± SD of 5 animals. (E) Mouse corneal micropocket assay was performed as described in Methods. Arrows indicate the site where pellets were implanted. Neovascular area (mm2) was determined using NIH Image. Values are expressed as mean ± SD of 5 eyes. (F) CAM assay using adenovirus vectors was performed as described in Methods. The nylon mesh containing adenovirus/Matrigel mixture was placed on the peripheral zone of the CAM, where vascular structure would not appear (left panels). Four days after adenovirus infection, vascular formation was evaluated by macroscopic observation (right). Arrowheads indicate the site where the nylon mesh was placed. *P < 0.05, **P < 0.01.