title: PECAM-targeted delivery of SOD inhibits endothelial inflammatory response.
abstract: Elevated generation of reactive oxygen species (ROS) by endothelial enzymes, including NADPH-oxidase, is implicated in vascular oxidative stress and endothelial proinflammatory activation involving exposure of vascular cell adhesion molecule-1 (VCAM-1). Catalase and superoxide dismutase (SOD) conjugated with antibodies to platelet/endothelial cell adhesion molecule 1 (PECAM-1) bind specifically to endothelium and inhibit effects of corresponding ROS, H(2)O(2), and superoxide anion. In this study, anti-PECAM/SOD, but not anti-PECAM/catalase or nontargeted enzymes, including polyethylene glycol (PEG)-SOD, inhibited 2- to 3-fold VCAM expression caused by tumor necrosis factor (TNF), interleukin-1β, and lipopolysaccharide. Anti- PECAM/SOD, but not nontargeted counterparts, accumulated in vascular endothelium after intravenous injection, localized in endothelial endosomes, and inhibited by 70% lipopolysaccharide-caused VCAM-1 expression in mice. Anti-PECAM/SOD colocalized with EEA-1-positive endothelial vesicles and quenched ROS produced in response to TNF. Inhibitors of NADPH oxidase and anion channel ClC3 blocked TNF-induced VCAM expression, affirming that superoxide produced and transported by these proteins, respectively, mediates inflammatory signaling. Anti-PECAM/SOD abolished VCAM expression caused by poly(I:C)-induced activation of toll-like receptor 3 localized in intracellular vesicles. These results directly implicate endosomal influx of superoxide in endothelial inflammatory response and suggest that site-specific interception of this signal attained by targeted delivery of anti-PECAM/SOD into endothelial endosomes may have anti-inflammatory effects.
authors.affiliation: Institute for Environmental Medicine, Department of Pharmacology, University of Pennsylvania School of Medicine, 1 John Morgan Bldg., 3620 Hamilton Walk, Philadelphia, PA 19104-6068, USA.
authors.forename: Vladimir V
authors.initials: VV
authors.lastname: Shuvaev
authors.forename: Jingyan
authors.initials: J
authors.lastname: Han
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Yu
authors.forename: Shaohui
authors.initials: S
authors.lastname: Huang
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Hawkins
authors.forename: Muniswamy
authors.initials: M
authors.lastname: Madesh
authors.forename: Marian
authors.initials: M
authors.lastname: Nakada
authors.forename: Vladimir R
authors.initials: VR
authors.lastname: Muzykantov
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D015153:Blotting, Western; D002460:Cell Line; D018118:Chloride Channels / Q000235:genetics / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D018796:Immunoconjugates / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D007275:Injections, Intravenous; D053583:Interleukin-1beta / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008168:Lung / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism; D008856:Microscopy, Fluorescence; D019255:NADPH Oxidases / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology*; D034622:RNA Interference; D013482:Superoxide Dismutase / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D013481:Superoxides / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018118:Chloride Channels; C115376:ClC-3 channel; D018796:Immunoconjugates; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D013481:Superoxides; D013482:Superoxide Dismutase; D019255:NADPH Oxidases