Soluble urokinase receptor conjugated to carrier red blood cells binds latent pro-urokinase and alters its functional profile.
Coupling plasminogen activators to carrier red blood cells (RBC) prolongs their life-time in the circulation and restricts extravascular side effects, thereby allowing their utility for short-term thromboprophylaxis. Unlike constitutively active plasminogen activators, single chain urokinase plasminogen activator (scuPA) is activated by plasmin proteolysis or binding to its receptor, uPAR. In this study we conjugated recombinant soluble uPAR (suPAR) to rat RBC, forming RBC/suPAR complex. RBC carrying suPAR circulated in rats similarly to naïve RBC and markedly prolonged the circulation time of suPAR. RBC/suPAR carrying approximately 3x10(4) suPAR molecules per RBC specifically bound up to 2x10(4) molecules of scuPA, retained approximately 75% of scuPA-binding capacity after circulation in rats and markedly altered the functional profile of bound scuPA. RBC carrying directly conjugated scuPA adhered to endothelial cells, while showing no appreciable fibrinolytic activity. In contrast, RBC/suPAR loaded with scuPA did not exhibit increased adhesion to endothelium, while effectively dissolving fibrin clots. This molecular design, capitalizing on unique biological features of the interaction of scuPA with its receptor, provides a promising modality to deliver a pro-drug for prevention of thrombosis.
Murciano
Juan-Carlos
JC
Centro Nacional de Investigaciones Cardiovasculares, Madrid Spain.
Higazi
Abd Al-Roof
AA
Cines
Douglas B
DB
Muzykantov
Vladimir R
VR
D000818:Animals; D019921:Biotinylation; D002448:Cell Adhesion; D002478:Cells, Cultured; D002626:Chemistry, Pharmaceutical; D004337:Drug Carriers; D004339:Drug Compounding; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D017707:Erythrocyte Transfusion; D004912:Erythrocytes / Q000378:metabolism*; D005342:Fibrinolysis / Q000187:drug effects; D005343:Fibrinolytic Agents / Q000097:blood / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D011355:Prodrugs / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D051381:Rats; D055293:Receptors, Urokinase Plasminogen Activator / Q000097:blood* / Q000737:chemistry; D011994:Recombinant Proteins / Q000097:blood; D015912:Thrombolytic Therapy; D014568:Urokinase-Type Plasminogen Activator / Q000097:blood / Q000737:chemistry / Q000494:pharmacology*
D004337:Drug Carriers; D005343:Fibrinolytic Agents; D011355:Prodrugs; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator