VEGFR2 and Src Kinase Inhibitors Suppress Andes Virus-Induced Endothelial Cell Permeability
VEGFR2 and Src Kinase Inhibitors Suppress Andes Virus-Induced Endothelial Cell Permeability
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DOI：
10.1128/jvi.02319-10
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发表时间：
2011-03-01
影响因子：
5.4
通讯作者：
Mackow, Erich R.
中科院分区：
文献类型：
--
作者：
Gorbunova, Elena E.;Gavrilovskaya, Irina N.;Mackow, Erich R.
汉坦病毒主要感染人内皮细胞，在没有细胞溶解的情况下，引起血管通透性增加引起的两种疾病。安第斯病毒（ANDV）引起高度致命的急性肺水肿称为汉坦病毒肺综合征（HPS）。ANDV感染通过增加由VEGFR2-Src-VE-cadherin通路指导的信号反应，从而增强内皮细胞对血管内皮生长因子（VEGF）的通透性，该通路指导粘附连接（AJ）的分解。本研究表明，抑制途径特异性VEGFR2和Src家族激酶（SFKs）可阻断andv诱导的内皮细胞通透性。在andv感染的内皮细胞中，小干扰RNA （siRNA）敲低Src导致内皮细胞通透性与siRNA对照组相比降低了70%。这一发现表明，现有的fda批准的小分子激酶抑制剂可能类似地阻断andv诱导的通透性。VEGFR2激酶抑制剂pazopanib以及SFK抑制剂达沙替尼、PP1、博舒替尼和Src抑制剂1显著抑制andv诱导的内皮细胞通透性。与其激酶抑制浓度一致，达沙替尼、PP1和帕唑帕尼分别在1、10和100纳摩尔50%抑制浓度（IC(50)s）下抑制andv诱导的通透性。我们进一步证明，达沙替尼和帕唑帕尼阻断了andv感染内皮细胞AJs中VE-cadherin的分离，阻断率为90%。这些发现表明，VEGFR2和Src激酶是治疗性降低andv诱导的内皮细胞通透性的潜在靶点，从而降低HPS期间的毛细血管通透性。由于VEGFR2和SFK抑制剂的功能已经得到了很好的定义，FDA也批准了它们的临床应用，因此这些发现合理化了它们在减少HPS疾病方面的疗效评估。内皮细胞屏障功能被许多病毒破坏，导致出血性、水肿或神经系统疾病，因此，我们的研究结果表明，应该考虑使用VEGFR2和SFK抑制剂来调节被其他病毒病原体改变的内皮细胞屏障功能。
Hantaviruses predominantly infect human endothelial cells and, in the absence of cell lysis, cause two diseases resulting from increased vascular permeability. Andes virus (ANDV) causes a highly lethal acute pulmonary edema termed hantavirus pulmonary syndrome (HPS). ANDV infection enhances the permeability of endothelial cells in response to vascular endothelial growth factor (VEGF) by increasing signaling responses directed by the VEGFR2-Src-VE-cadherin pathway, which directs adherens junction (AJ) disassembly. Here we demonstrate that inhibiting pathway-specific VEGFR2 and Src family kinases (SFKs) blocks ANDV-induced endothelial cell permeability. Small interfering RNA (siRNA) knockdown of Src within ANDV-infected endothelial cells resulted in an similar to 70% decrease in endothelial cell permeability compared to that for siRNA controls. This finding suggested that existing FDA-approved small-molecule kinase inhibitors might similarly block ANDV-induced permeability. The VEGFR2 kinase inhibitor pazopanib as well as SFK inhibitors dasatinib, PP1, bosutinib, and Src inhibitor 1 dramatically inhibited ANDV-induced endothelial cell permeability. Consistent with their kinase-inhibitory concentrations, dasatinib, PP1, and pazopanib inhibited ANDV-induced permeability at 1, 10, and 100 nanomolar 50% inhibitory concentrations (IC(50)s), respectively. We further demonstrated that dasatinib and pazopanib blocked VE-cadherin dissociation from the AJs of ANDV-infected endothelial cells by >90%. These findings indicate that VEGFR2 and Src kinases are potential targets for therapeutically reducing ANDV-induced endothelial cell permeability and, as a result, capillary permeability during HPS. Since the functions of VEGFR2 and SFK inhibitors are already well defined and FDA approved for clinical use, these findings rationalize their therapeutic evaluation for efficacy in reducing HPS disease. Endothelial cell barrier functions are disrupted by a number of viruses that cause hemorrhagic, edematous, or neurologic disease, and as a result, our findings suggest that VEGFR2 and SFK inhibitors should be considered for regulating endothelial cell barrier functions altered by additional viral pathogens.