Endothelial-derived von Willebrand factor accelerates fibrin clotting within engineered microvessels.
Endothelial-derived von Willebrand factor accelerates fibrin clotting within engineered microvessels.
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内皮源性血管性血友病因子可加速工程微血管内的纤维蛋白凝固
DOI：
10.1111/jth.15714
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发表时间：
2022-07
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影响因子：
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中科院分区：
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血管性血友病因子（VWF）通常与原发性止血和富血小板动脉血栓形成有关，但最近也与纤维蛋白凝固和静脉血栓形成有关。纤维蛋白和VWF之间的直接相互作用可能介导这些过程，虽然以前的报告是相互矛盾的。我们结合了两个互补的平台来表征VWF-纤维蛋白（原）相互作用，并确定其潜在的生理意义。工程化微血管内衬人内皮细胞，在流动下培养，并活化以释放VWF并形成经腔VWF纤维。然后灌注纤维蛋白原、纤维蛋白单体或聚合纤维蛋白，并评价与VWF的相互作用。凝血酶和纤维蛋白原灌注到活的与多聚甲醛固定的微血管和跨微血管的压降进行监测。单独地，使用全内反射荧光（TIRF）显微镜在单分子水平上评估蛋白质与系留VWF的结合。在微血管内，VWF纤维与聚合纤维蛋白共定位，但不与纤维蛋白原共定位。TIRF显微镜显示，在一个微流体流动室中，在一系列的剪切速率和蛋白质浓度范围内，VWF和纤维蛋白原或纤维蛋白单体之间没有共定位。凝血酶介导的活微血管内的纤维蛋白聚合触发内皮VWF释放，与具有惰性内皮的固定血管相比，增加微血管阻塞的速率和量。我们没有发现纤维蛋白（原）和VWF之间在单分子水平的特异性结合。尽管如此，我们的研究结果表明，在凝血过程中内皮VWF的快速释放可能为纤维蛋白聚合提供物理支持并加速血栓形成。这种相互作用可能是人类血栓性疾病的理解和治疗的根本重要性。
Von Willebrand factor (VWF) is classically associated with primary hemostasis and platelet-rich arterial thromboses, but recently has also been implicated in fibrin clotting and venous thrombosis. Direct interaction between fibrin and VWF may mediate these processes, although prior reports are conflicting. We combined two complementary platforms to characterize VWF-fibrin(ogen) interactions and identify their potential physiologic significance. Engineered microvessels were lined with human endothelial cells, cultured under flow, and activated to release VWF and form transluminal VWF fibers. Fibrinogen, fibrin monomers, or polymerizing fibrin were then perfused, and interactions with VWF evaluated. Thrombin and fibrinogen were perfused into living versus paraformeldahyde-fixed microvessels and the pressure drop across microvessels monitored. Separately, protein binding to tethered VWF was assessed on a single-molecule level using total internal reflection fluorescence (TIRF) microscopy. Within microvessels, VWF fibers colocalized with polymerizing fibrin, but not fibrinogen. TIRF microscopy showed no colocalization between VWF and fibrinogen or fibrin monomers in a microfluidic flow chamber across a range of shear rates and protein concentrations. Thrombin-mediated fibrin polymerization within living microvessels triggered endothelial VWF release, increasing the rate and amount of microvessel obstruction compared to fixed vessels with an inert endothelium. We did not identify specific binding between fibrin(ogen) and VWF at a single-molecule level. Despite this, our results suggest that rapid release of endothelial VWF during clotting may provide a physical support for fibrin polymerization and accelerate thrombosis. This interaction may be of fundamental importance for the understanding and treatment of human thrombotic disease.
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影响因子：
4
作者：
Hoffman, M
通讯作者：
Hoffman, M
影响因子：
7.5
作者：
Kragh, Thorsten;Napoleone, Marina;Reininger, Armin J.
通讯作者：
Reininger, Armin J.
影响因子：
15.9
作者：
LOSCALZO, J;INBAL, A;HANDIN, RI
通讯作者：
HANDIN, RI
影响因子：
20.3
作者：
ENDENBURG, SC;HANTGAN, RR;DEGROOT, PG
通讯作者：
DEGROOT, PG
DOI：
10.1111/j.1538-7836.2011.04544.x
发表时间：
2012-01
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Brill A;Fuchs TA;Savchenko AS;Thomas GM;Martinod K;De Meyer SF;Bhandari AA;Wagner DD
通讯作者：
Wagner DD