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STAT1 activation by venous malformations mutant Tie2-R849W antagonizes VEGF-A-mediated angiogenic response partly via reduced bFGF production

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Abstract

A missense mutation from arginine to tryptophan at residue 849 in the kinase domain of Tie2 (Tie2-R849W) is commonly identified in familial venous malformations. The mechanistic action of Tie2-R849W variant expression on angiogenic cascades including smooth muscle cell recruitment, however, remains elusive. To avoid confounding factors from endogenous Tie2 expression, Tie2-depleted endothelial cells (ECs) were used to study the effects of ectopic shRNA-resistant Tie2 variant expression, Tie2-WT* and Tie2-R849W*, on vascular cell proliferation, migration, tube formation, and smooth muscle cell (SMC) recruitment. Tie2-R849W* induced STAT1 phosphorylation at Tyr701. Tie2-R849W*-expressing cells had reduced ability to migrate and form tubes on Matrigel than their wildtype counterparts. STAT1 phosphorylation attenuated VEGF-A-induced STAT3 tyrosine phosphorylation in Tie2-R849W*-expressing HUVECs. The induced STAT1 activation also decreased VEGF-A-induced bFGF mRNA expression by competing with activated STAT3 for a direct binding to the consensus STAT-binding site at positions −997 to −989 bp from transcription start site in the bFGF promoter. Depleting STAT1 expression rescued the inability of Tie2-R849W expression to mediate angiogenesis. Moreover, bFGF neutralization or constitutive STAT1 activation, reminiscence of Tie2-R849W* expression, suppressed the smooth muscle cell recruiting ability of endothelial conditioned medium. This work reveals an anti-angiogenic role of STAT1 activation that acts in Tie2-R849W-expressing ECs to impair VEGF-A-mediated STAT3 signaling, bFGF production, and smooth muscle cell recruitment. A balancing activity of STAT1 and STAT3 may be important for Tie2-mediated vascular homeostasis.

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Acknowledgments

This work was supported by National Science Council (98-2320-B-006-034-MY3, 99-2628-B-006-017-MY3, and 100-2325-B006-005) and Department of Health (DOH-101-TD-C-111-003) to Wu LW. A fellowship for establishing centers of excellence for cancer research from Department of Health, Executive Yuan in Taiwan (DOH100-TD-C-111-003) was awarded to Huang YH. All shRNA plasmids were obtained from the National RNAi Core Facility at the Institute of Molecular Biology/Genomic Research Center, Academia Sinica, supported by the National Research Program for Genomic Medicine Grants by National Science Council (NSC 97-B-3112-B-001-016). All the authors state no conflict of interest.

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Authors and Affiliations

  1. Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, 1 University Rd., Tainan, 701, Taiwan, ROC

    Yi-Hsien Huang & Li-Wha Wu

  2. Graduate Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, 1 University Rd., Tainan, 701, Taiwan, ROC

    Wu-Chou Su

  3. Institute of Molecular Medicine, College of Medicine, National Cheng Kung University, 1 University Rd., Tainan, 701, Taiwan, ROC

    Yi-Wen Chen & Li-Wha Wu

  4. Division of Urogynecology and Pelvic Floor Reconstruction, Department of Obstetrics & Gynecology, Chi Mei Medical Center, 901 Zhonghua Rd., Yongkang District, Tainan, 710, Taiwan, ROC

    Ming-Ping Wu

  5. Department of Obstetrics & Gynecology, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei, 110, Taiwan, ROC

    Ming-Ping Wu

  6. Section of Plastic and Reconstructive Surgery, Department of Surgery, National Cheng Kung University Hospital, 138 Sheng-Li Rd., Tainan, 704, Taiwan, ROC

    Shin-Chen Pan

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Huang, YH., Wu, MP., Pan, SC. et al. STAT1 activation by venous malformations mutant Tie2-R849W antagonizes VEGF-A-mediated angiogenic response partly via reduced bFGF production. Angiogenesis 16, 207–222 (2013). https://doi.org/10.1007/s10456-012-9313-x

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