Abstract Number: OC 02.2
Meeting: ISTH 2020 Congress
Background: Plasma fibrinogen concentration correlates with cardiovascular disease (CVD) event risk, but a causative role for fibrinogen has not been established, and it is unclear if lowering fibrinogen levels could be beneficial for CVD. A small molecule modulator of fibrinogen production could be used to investigate this.
Aims: We aimed to identify chemical entities capable of changing fibrinogen production and test their impact on experimental thrombosis.
Methods: 1280 compounds were screened for their ability to alter fibrinogen production in hepatocyte-derived HepG2 cells. The screen was performed three times and lead compounds tested for dose-responsiveness and efficacy prior to validation in human HuH7 and murine AML12 cells. A panel of compounds were assessed for fibrinogen modulation and effects on laser-induced venous thrombosis assays in zebrafish larvae. Gene expression, proteomics, and morpholino-based mRNA knockdowns were used to investigate mechanisms-of-action.
Results: Our chemical screen identified anthralin and all-trans retinoic acid (RA) 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 the opposite effects. Anthralin had little effect on fibrinogen mRNA in zebrafish larvae but, using a proteomic scan of anthralin-treated cells and zebrafish, we detected reduced representation of proteins with secretory signal peptides, suggesting a mechanism whereby anthralin affects proteins via the canonical secretory pathway. Treatment of cultured cells or zebrafish larvae with RA increased steady-state fibrinogen mRNA. Using a morpholino oligonucleotide to deplete zebrafish fibrinogen mRNA, we correlated the shortening of venous thrombosis times with RA and fibrinogen protein levels.
Conclusions: We identified anthralin as a fibrinogen-lowering small molecule and are now testing its effects on fibrinogen levels and hemostasis in mice. Anthralin, or an optimized derivative with improved selectivity, could be used to assess the protective effect of lowering plasma fibrinogen on cardiovascular disease-related event risk.
To cite this abstract in AMA style:Vilar R, Lukowski SW, Garieri M, Di Sanza C, Neerman-Arbez M, Fish RJ. Chemical Modulation of Fibrinogen Production and its Impact on Venous Thrombosis [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/chemical-modulation-of-fibrinogen-production-and-its-impact-on-venous-thrombosis/. Accessed February 28, 2024.
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