title: Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents.
abstract: 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.
authors.affiliation: Division of Hemostasis and Thrombosis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Reema
authors.initials: R
authors.lastname: Jasuja
authors.forename: Freda H
authors.initials: FH
authors.lastname: Passam
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Kennedy
authors.forename: Sarah H
authors.initials: SH
authors.lastname: Kim
authors.forename: Lotte
authors.initials: L
authors.lastname: van Hessem
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.forename: Sheryl R
authors.initials: SR
authors.lastname: Bowley
authors.forename: Sucharit S
authors.initials: SS
authors.lastname: Joshi
authors.forename: James R
authors.initials: JR
authors.lastname: Dilks
authors.forename: Bruce
authors.initials: B
authors.lastname: Furie
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Furie
authors.forename: Robert
authors.initials: R
authors.lastname: Flaumenhaft
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005337:Fibrin / Q000235:genetics / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D010974:Platelet Aggregation / Q000187:drug effects*; D019704:Protein Disulfide-Isomerases / Q000009:adverse effects / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000494:pharmacology; D011994:Recombinant Proteins / Q000009:adverse effects / Q000037:antagonists & inhibitors / Q000235:genetics / Q000494:pharmacology; D012431:Rutin / Q000494:pharmacology*; D013927:Thrombosis / Q000139:chemically induced / Q000188:drug therapy* / Q000201:enzymology
chemical_list: D004791:Enzyme Inhibitors; D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D012431:Rutin; D005337:Fibrin; D019704:Protein Disulfide-Isomerases