Angiopoietin-like protein 2 regulates endothelial colony forming cell vasculogenesis.
Angiopoietin-like protein 2 regulates endothelial colony forming cell vasculogenesis.
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DOI：
10.1007/s10456-014-9423-8
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发表时间：
2014-07
期刊：
影响因子：
9.8
通讯作者：
Yoder, Mervin C.
中科院分区：
文献类型：
--
作者：
Richardson, Matthew R.;Robbins, Emilie P.;Vemula, Sasidhar;Critser, Paul J.;Whittington, Catherine;Voytik-Harbin, Sherry L.;Yoder, Mervin C.
据报道，血管生成素样2 （ANGPTL2）可诱导发芽血管生成；然而，它在血管发生中的作用，内皮细胞（EC）的新生流明化，仍未被探索。我们试图通过siRNA介导的抑制ANGPTL2基因表达来研究ANGPTL2在调节人脐带血来源的内皮集落形成细胞（ECFC）血管生成中的潜在作用。我们发现ANGPTL2减少的ecfc在体外管腔面积减少了3倍，而在血管发生的三维胶原实验中，向ecfc添加外源性ANGPTL2蛋白结构域导致管腔形成增加。通过敲除ANGPTL2 (KD)， ECFC的迁移减少了36%，但增殖和凋亡不受影响。我们随后发现，ANGPTL2 KD导致JNK（而非ERK1/2）磷酸化降低，而MT1-MMP（已知受JNK调控，是EC迁移和3D管腔形成的关键调节剂）的表达在体外来源于ANGPTL2沉默的ecfc的流明化结构中降低。用JNK抑制剂或外源性rhTIMP-3 （MT1-MMP活性抑制剂）处理3D胶原基质中的ecfc，结果与ANGPTL2 KD观察到的血管腔形成减少的表型相似，而刺激JNK活性则增加了血管发生。基于基因沉默、药理学、细胞和生化方法，我们得出结论，ANGPTL2可能通过其对迁移的影响，部分通过激活JNK和增加MT1-MMP表达，积极调节ECFC血管管腔的形成。
Angiopoietin-like 2 (ANGPTL2) has been reported to induce sprouting angiogenesis; however, its role in vasculogenesis, the de novo lumenization of endothelial cells (EC), remains unexplored. We sought to investigate the potential role of ANGPTL2 in regulating human cord blood derived endothelial colony forming cell (ECFC) vasculogenesis through siRNA mediated inhibition of ANGPTL2 gene expression. We found that ECFCs in which ANGPTL2 was diminished displayed a 3-fold decrease in in vitro lumenal area whereas addition of exogenous ANGPTL2 protein domains to ECFCs lead to increased lumen formation within a 3 dimensional collagen assay of vasculogenesis. ECFC migration was attenuated by 36% via ANGPTL2 knockdown (KD) although proliferation and apoptosis were not affected. We subsequently found that JNK, but not ERK1/2, phosphorylation was decreased upon ANGPTL2 KD, and expression of MT1-MMP, known to be regulated by JNK and a critical regulator of EC migration and 3D lumen formation, was decreased in lumenized structures in vitro derived from ANGPTL2 silenced ECFCs. Treatment of ECFCs in 3D collagen matrices with either a JNK inhibitor or exogenous rhTIMP-3 (an inhibitor of MT1-MMP activity) resulted in a similar phenotype of decreased vascular lumen formation as observed with ANGPTL2 KD, whereas stimulation of JNK activity increased vasculogenesis. Based on gene silencing, pharmacologic, cellular, and biochemical approaches, we conclude that ANGPTL2 positively regulates ECFC vascular lumen formation likely through its effects on migration and in part by activating JNK and increasing MT1-MMP expression.
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影响因子：
20.3
作者：
Sacharidou, Anastasia;Koh, Wonshill;Davis, George E.
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