Extracellular Vesicles Activate a CD36-Dependent Signaling Pathway to Inhibit Microvascular Endothelial Cell Migration and Tube Formation.
Extracellular Vesicles Activate a CD36-Dependent Signaling Pathway to Inhibit Microvascular Endothelial Cell Migration and Tube Formation.
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DOI：
10.1161/atvbaha.115.307085
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发表时间：
2016-03
期刊：
影响因子：
--
通讯作者：
Silverstein RL
中科院分区：
文献类型：
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作者：
Ramakrishnan DP;Hajj-Ali RA;Chen Y;Silverstein RL
关于细胞来源的细胞外囊泡（EV）（活化或凋亡期间从各种细胞类型脱落的≤1μm囊泡）对微血管内皮细胞（MVEC）信号传导的影响的文献相互矛盾。血小板反应蛋白-1和相关蛋白通过CD 36诱导MVEC中的抗血管生成信号。CD 36通过暴露在其表面上的磷脂酰丝氨酸（PS）结合EV，但这种相互作用对MVEC功能的影响尚不清楚。我们假设EV通过CD 36抑制血管生成MVEC功能。在体外基质胶试验和Boyden小室试验中，从各种细胞类型中体外产生的EV或从血浆中分离的EV抑制MVEC管形成和EC迁移。来源于相同细胞的外来体不具有抑制活性。抑制迁移需要EC表达CD 36。在小鼠体内基质胶塞测定EV抑制细胞迁移到基质胶塞在野生型，但不是在cd 36空动物。膜联蛋白V，一种阴离子磷脂结合蛋白，当与EV孵育时，部分逆转了对迁移的抑制，表明PS依赖性效应。EV暴露诱导活性氧（ROS）的产生在MVEC中的NADPH氧化酶和Src家族激酶依赖的方式和他们的抑制夹竹桃黄素和PP 2，分别部分逆转EV介导的抑制迁移。Annexin V部分逆转EV诱导的ROS在鼠CD 36 cDNA转染的HVUEC中的产生，但在CD 36阴性的HUVEC中不逆转。这些研究建立了EV对EC促血管生成反应的一般抑制作用，并确定了EV抑制MVEC迁移和管形成的CD 36介导的机制途径。
Literature on the effect of cell-derived extracellular vesicles (EV), ≤1μm vesicles shed from various cell types during activation or apoptosis, on microvascular endothelial cell (MVEC) signaling is conflicting. Thrombospondin-1 and related proteins induce anti-angiogenic signals in MVEC via CD36. CD36 binds EV via phosphatidylserine (PS) exposed on their surface but the effects of this interaction on MVEC functions are not known. We hypothesized that EV would inhibit angiogenic MVEC functions via CD36. EV generated in vitro from various cell types or isolated from plasma inhibited MVEC tube formation in in vitro matrigel assays and EC migration in Boyden chamber assays. Exosomes derived from the same cells did not have inhibitory activity. Inhibition of migration required EC expression of CD36. In mouse in vivo matrigel plug assays EV inhibited cell migration into matrigel plugs in wild type but not in cd36 null animals. Annexin V, an anionic phospholipid binding protein, when incubated with EV partially reversed inhibition of migration, suggesting a PS-dependent effect. EV exposure induced reactive oxygen species (ROS) generation in MVEC in a NADPH oxidase and Src family kinase dependent manner and their inhibition by apocynin and PP2, respectively, partially reversed the EV-mediated inhibition of migration. Annexin V partially reversed EV-induced ROS generation in murine CD36 cDNA transfected HVUEC but not in CD36 negative HUVEC. These studies establish a general inhibitory effect of EV on EC pro-angiogenic responses and identify a CD36-mediated mechanistic pathway through which EV inhibit MVEC migration and tube formation.