title: Dual targeting of therapeutics to endothelial cells: collaborative enhancement of delivery and effect.
abstract: Anchoring pharmacologic agents to the vascular lumen has the potential to modulate critical processes at the blood-tissue interface, avoiding many of the off-target effects of systemically circulating agents. We report a novel strategy for endothelial dual targeting of therapeutics, which both enhances drug delivery and enables targeted agents to partner enzymatically to generate enhanced biologic effect. Based on the recent discovery that paired antibodies directed to adjacent epitopes of platelet endothelial cell adhesion molecule (PECAM)-1 stimulate each other's binding, we fused single-chain fragments (scFv) of paired anti-mouse PECAM-1 antibodies to recombinant murine thrombomodulin (TM) and endothelial protein C receptor (EPCR), endothelial membrane proteins that partner in activation of protein C (PC). scFv/TM and scFv/EPCR bound to mouse endothelial PECAM-1 with high affinity (EC50 1.5 and 3.8 nM, respectively), and codelivery induced a 5-fold increase in PC activation not seen when TM and EPCR are anchored to distinct cell adhesion molecules. In a mouse model of acute lung injury, dual targeting reduces both the expression of lung inflammatory markers and trans-endothelial protein leak by as much as 40%, as compared to either agent alone. These findings provide proof of principle for endothelial dual targeting, an approach with numerous potential biomedical applications.
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Colin F
authors.initials: CF
authors.lastname: Greineder
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Brenza
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Ronald
authors.initials: R
authors.lastname: Carnemolla
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Sergei
authors.initials: S
authors.lastname: Zaitsev
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Elizabeth D
authors.initials: ED
authors.lastname: Hood
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Pan
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Bi-Sen
authors.initials: BS
authors.lastname: Ding
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Charles T
authors.initials: CT
authors.lastname: Esmon
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA.
authors.forename: Ann Marie
authors.initials: AM
authors.lastname: Chacko
authors.affiliation: *Department of Pharmacology, Center for Targeted Therapeutics and Translational Nanomedicine, Institute for Translational Medicine and Therapeutics, and Departments of Radiology and Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; Department of Genetic Medicine, Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA; and Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA muzykant@mail.med.upenn.edu.
authors.forename: Vladimir R
authors.initials: VR
authors.lastname: Muzykantov
mesh_terms: D055371:Acute Lung Injury / Q000188:drug therapy / Q000378:metabolism; D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D016503:Drug Delivery Systems / Q000379:methods; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D000939:Epitopes / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D004364:Pharmaceutical Preparations / Q000008:administration & dosage*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011486:Protein C / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D018180:Thrombomodulin / Q000378:metabolism
chemical_list: D015815:Cell Adhesion Molecules; D000939:Epitopes; D004364:Pharmaceutical Preparations; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011486:Protein C; D011956:Receptors, Cell Surface; D018180:Thrombomodulin
keywords: EPCR; PECAM-1; drug delivery; thrombomodulin