Department of Genetic Medicine and Development, University of Geneva Faculty of Medicine, Geneva, Switzerland.
Institute for Molecular Bioscience, The University of Queensland, Saint Lucia, Queensland, Australia.
Thromb Haemost. 2021 Apr;121(4):433-448. doi: 10.1055/s-0040-1718414. Epub 2020 Dec 10.
Thrombosis is a leading cause of morbidity and mortality. Fibrinogen, the soluble substrate for fibrin-based clotting, has a central role in haemostasis and thrombosis and its plasma concentration correlates with cardiovascular disease event risk and a prothrombotic state in experimental models. We aimed to identify chemical entities capable of changing fibrinogen production and test their impact on experimental thrombosis. A total of 1,280 bioactive compounds were screened for their ability to alter fibrinogen production by hepatocyte-derived cancer cells and a selected panel was tested in zebrafish larvae. Anthralin and all- retinoic acid (RA) were identified as fibrinogen-lowering and fibrinogen-increasing moieties, respectively. In zebrafish larvae, anthralin prolonged laser-induced venous- occlusion times and reduced thrombocyte accumulation at injury sites. RA had opposite effects. Treatment with RA, a nuclear receptor ligand, increased fibrinogen mRNA levels. Using an antisense morpholino oligonucleotide to deplete zebrafish fibrinogen, we correlated a shortening of laser-induced venous thrombosis times with RA treatment and fibrinogen protein levels. Anthralin had little effect on fibrinogen mRNA in zebrafish larvae, despite leading to lower detectable fibrinogen. Therefore, we made a proteomic scan of anthralin-treated cells and larvae. A reduced representation of proteins linked to the canonical secretory pathway was detected, suggesting that anthralin affects protein secretion. In summary, we found that chemical modulation of fibrinogen levels correlates with measured effects on experimental venous thrombosis and could be investigated as a therapeutic avenue for thrombosis prevention.
血栓形成是发病率和死亡率的主要原因。纤维蛋白原是纤维蛋白为基础的凝血的可溶性底物，在止血和血栓形成中起核心作用，其血浆浓度与心血管疾病事件风险和实验模型中的促血栓形成状态相关。我们旨在确定能够改变纤维蛋白原产生的化学实体，并测试它们对实验性血栓形成的影响。总共筛选了 1280 种生物活性化合物，以鉴定其改变肝癌细胞产生纤维蛋白原的能力，并在斑马鱼幼虫中测试了选定的化合物。蒽林和全反式视黄酸（RA）分别被鉴定为降低纤维蛋白原和增加纤维蛋白原的部分。在斑马鱼幼虫中，蒽林延长了激光诱导的静脉阻塞时间，并减少了损伤部位血小板的聚集。RA 则有相反的效果。用核受体配体 RA 治疗可增加纤维蛋白原 mRNA 水平。用抗纤维蛋白原的反义 morpholino 寡核苷酸处理，我们将激光诱导的静脉血栓形成时间的缩短与 RA 治疗和纤维蛋白原蛋白水平相关联。尽管蒽林导致可检测的纤维蛋白原降低，但对斑马鱼幼虫的纤维蛋白原 mRNA 几乎没有影响。因此，我们对蒽林处理的细胞和幼虫进行了蛋白质组扫描。检测到与经典分泌途径相关的蛋白质的代表性降低，表明蒽林影响蛋白质分泌。总之，我们发现纤维蛋白原水平的化学调节与对实验性静脉血栓形成的测量影响相关，可作为预防血栓形成的治疗途径进行研究。
