Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents

• Published in: The Journal of clinical investigation Vol. 122; no. 6; pp. 2104 - 2113
• Main Authors: Jasuja, Reema, Passam, Freda H, Kennedy, Daniel R, Kim, Sarah H, van Hessem, Lotte, Lin, Lin, Bowley, Sheryl R, Joshi, Sucharit S, Dilks, James R, Furie, Bruce, Furie, Barbara C, Flaumenhaft, Robert
• Format: Journal Article
• Language: English
• Published: United States American Society for Clinical Investigation 01.06.2012
• Subjects: Animals; Biomedical research; Blood clots; Blood Platelets - metabolism; Endoplasmic reticulum; Enzyme inhibitors; Enzyme Inhibitors - pharmacology; Enzymes; Fibrin - genetics; Fibrin - metabolism; Fibrinolytic Agents - pharmacology; Humans; Identification and classification; Mice; Platelet Aggregation - drug effects; Protein Disulfide-Isomerases - adverse effects; Protein Disulfide-Isomerases - antagonists & inhibitors; Protein Disulfide-Isomerases - genetics; Protein Disulfide-Isomerases - pharmacology; Proteins; Recombinant Proteins - adverse effects; Recombinant Proteins - antagonists & inhibitors; Recombinant Proteins - genetics; Recombinant Proteins - pharmacology; Rutin - pharmacology; Thrombolytic drugs; Thrombosis; Thrombosis - chemically induced; Thrombosis - drug therapy; Thrombosis - enzymology; United States
• ISSN: 0021-9738; 1558-8238
• DOI: 10.1172/JCI61228

Thrombosis, or blood clot formation, and its sequelae remain a leading cause of morbidity and mortality, and recurrent thrombosis is common despite current optimal therapy. Protein disulfide isomerase (PDI) is an oxidoreductase that has recently been shown to participate in thrombus formation. While currently available antithrombotic agents inhibit either platelet aggregation or fibrin generation, inhibition of secreted PDI blocks the earliest stages of thrombus formation, suppressing both pathways. Here, we explored extracellular PDI as an alternative target of antithrombotic therapy. A high-throughput screen identified quercetin-3-rutinoside as an inhibitor of PDI reductase activity in vitro. Inhibition of PDI was selective, as quercetin-3-rutinoside failed to inhibit the reductase activity of several other thiol isomerases found in the vasculature. Cellular assays showed that quercetin-3-rutinoside inhibited aggregation of human and mouse platelets and endothelial cell-mediated fibrin generation in human endothelial cells. Using intravital microscopy in mice, we demonstrated that quercetin-3-rutinoside blocks thrombus formation in vivo by inhibiting PDI. Infusion of recombinant PDI reversed the antithrombotic effect of quercetin-3-rutinoside. Thus, PDI is a viable target for small molecule inhibition of thrombus formation, and its inhibition may prove to be a useful adjunct in refractory thrombotic diseases that are not controlled with conventional antithrombotic agents.