Drug Safety Research and Evaluation, Research, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-Chome, Fujisawa, Kanagawa, 251-8555, Japan.
Sci Rep. 2024 Jun 19;14(1):14109. doi: 10.1038/s41598-024-64063-y.
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.
开发一种可靠的方法来预测血小板减少症在药物发现中至关重要。在这里，我们建立了一种使用微生理系统（MPS）的测定方法，以重现血小板聚集和黏附的体内机制。该测定方法突出了切应力对血小板聚集及其与血管内皮细胞相互作用的作用。使用平板摇床和 MPS 中的重力驱动流在搅拌而非静态条件下检测可溶性胶原诱导的血小板聚集。值得注意的是，在 MPS 中的重力驱动流下，血小板聚集物黏附在血管内皮细胞上，并且这种事件呈浓度依赖性增加。在比较富含血小板的血浆（PRP）和全血中可溶性胶原诱导的聚集活性时，在 PRP 和全血中分别观察到浓度为 30μg/mL 和 3μg/mL 时出现明显的血小板聚集形成。此外，寡核苷酸 ODN2395 诱导血小板聚集和黏附到血管内皮细胞。通过血小板上的糖蛋白 VI 介导血栓形成活性的 SYK 抑制，改善了该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和寡核苷酸诱导的。我们的评估系统部分重现了血管中的聚集机制，并有助于发现安全药物以降低血小板减少症的风险。
