Fibrinogen Interaction With Complement C3: A Potential Therapeutic Target To Reduce Thrombosis Risk

Complement C3 binds fibrinogen and compromises fibrin clot lysis, thereby enhancing the risk of thrombosis. We investigated the role of the fibrinogen-C3 interaction as a novel therapeutic target to reduce thrombosis risk by analyzing: (i) consistency in the fibrinolytic properties of C3; (ii) binding sites between fibrinogen and C3; and (iii) modulation of fibrin clot lysis by manipulating fibrinogen-C3 interactions. Purified fibrinogen and C3 from the same individuals (n=24) were used to assess inter-individual variability in the anti-fibrinolytic effects of C3. Microarray screening and molecular modeling evaluated C3 and fibrinogen interaction sites. Novel synthetic conformational proteins, termed affimers, were used to modulate the C3-fibrinogen interaction and fibrinolysis. C3 purified from patients with type 1 diabetes showed enhanced prolongation of fibrinolysis compared with healthy control protein (195 +/- 105 and 522 +/- 166 s, respectively; P=0.04), with consistent effects but a wider range (5-51% and 5-18% lysis prolongation, respectively). Peptide microarray screening identified two potential C3-fibrinogen interaction sites within the fibrinogen b chain (residues 424-433 and 435-445). One fibrinogen-binding affimer that was isolated displayed sequence identity with C3 in an exposed area of the protein. This affimer abolished C3induced prolongation of fibrinolysis (728 +/- 25.1 s to 632 +/- 23.7 s; P=0.005) and showed binding to fibrinogen in the same region that is involved in C3-fibrinogen interactions. Moreover, it shortened plasma clot lysis of patients with diabetes, cardiovascular disease or controls by 7-11%. C3 binds fibrinogen b-chain and disruption of the fibrinogen-C3 interaction using affimer proteins enhances fibrinolysis, which represents a potential novel tool to reduce thrombosis in high-risk individuals.