PDF(Pubmed)
Abstract：
Developing a reliable method to predict thrombocytopenia is imperative in drug discovery. Here, we establish an assay using a microphysiological system (MPS) to recapitulate the in-vivo mechanisms of platelet aggregation and adhesion. This assay highlights the role of shear stress on platelet aggregation and their interactions with vascular endothelial cells. Platelet aggregation induced by soluble collagen was detected under agitated, but not static, conditions using a plate shaker and gravity-driven flow using MPS. Notably, aggregates adhered on vascular endothelial cells under gravity-driven flow in the MPS, and this incident increased in a concentration-dependent manner. Upon comparing the soluble collagen-induced aggregation activity in platelet-rich plasma (PRP) and whole blood, remarkable platelet aggregate formation was observed at concentrations of 30 µg/mL and 3 µg/mL in PRP and whole blood, respectively. Moreover, ODN2395, an oligonucleotide, induced platelet aggregation and adhesion to vascular endothelial cells. SYK inhibition, which mediated thrombogenic activity via glycoprotein VI on platelets, ameliorated platelet aggregation in the system, demonstrating that the mechanism of platelet aggregation was induced by soluble collagen and oligonucleotide. Our evaluation system partially recapitulated the aggregation mechanisms in blood vessels and can contribute to the discovery of safe drugs to mitigate the risk of thrombocytopenia.
摘要：
开发一种可靠的方法来预测血小板减少症在药物发现中势在必行。这里,我们使用微生理系统（MPS）建立了一种测定法，以概括血小板聚集和粘附的体内机制。该测定强调了剪切应力对血小板聚集的作用及其与血管内皮细胞的相互作用。在搅拌下检测到可溶性胶原蛋白诱导的血小板聚集，但不是静态的，使用平板振动筛和使用MPS的重力驱动流的条件。值得注意的是,在重力驱动下，MPS中粘附在血管内皮细胞上的聚集体，这一事件以浓度依赖的方式增加。通过比较富含血小板的血浆（PRP）和全血中可溶性胶原蛋白诱导的聚集活性，在PRP和全血中30µg/mL和3µg/mL的浓度下观察到明显的血小板聚集体形成，分别。此外，ODN2395，一种寡核苷酸，诱导血小板聚集和粘附血管内皮细胞。SYK抑制,通过血小板上的糖蛋白VI介导血栓形成活性，改善系统中的血小板聚集，表明血小板聚集的机制是由可溶性胶原蛋白和寡核苷酸诱导的。我们的评估系统部分概述了血管中的聚集机制，并有助于发现安全的药物以减轻血小板减少症的风险。
