title: MicroRNA-147b regulates vascular endothelial barrier function by targeting ADAM15 expression.
abstract: A disintegrin and metalloproteinase15 (ADAM15) has been shown to be upregulated and mediate endothelial hyperpermeability during inflammation and sepsis. This molecule contains multiple functional domains with the ability to modulate diverse cellular processes including cell adhesion, extracellular matrix degradation, and ectodomain shedding of transmembrane proteins. These characteristics make ADAM15 an attractive therapeutic target in various diseases. The lack of pharmacological inhibitors specific to ADAM15 prompted our efforts to identify biological or molecular tools to alter its expression for further studying its function and therapeutic implications. The goal of this study was to determine if ADAM15-targeting microRNAs altered ADAM15-induced endothelial barrier dysfunction during septic challenge by bacterial lipopolysaccharide (LPS). An in silico analysis followed by luciferase reporter assay in human vascular endothelial cells identified miR-147b with the ability to target the 3' UTR of ADAM15. Transfection with a miR-147b mimic led to decreased total, as well as cell surface expression of ADAM15 in endothelial cells, while miR-147b antagomir produced an opposite effect. Functionally, LPS-induced endothelial barrier dysfunction, evidenced by a reduction in transendothelial electric resistance and increase in albumin flux across endothelial monolayers, was attenuated in cells treated with miR-147b mimics. In contrast, miR-147b antagomir exerted a permeability-increasing effect in vascular endothelial cells similar to that caused by LPS. Taken together, these data suggest the potential role of miR147b in regulating endothelial barrier function by targeting ADAM15 expression.
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Victor
authors.initials: V
authors.lastname: Chatterjee
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Richard S
authors.initials: RS
authors.lastname: Beard
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Jason J
authors.initials: JJ
authors.lastname: Reynolds
authors.affiliation: Department of Surgery, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Ricci
authors.initials: R
authors.lastname: Haines
authors.affiliation: Department of Surgery, University of California Davis School of Medicine, Sacramento, California, United States of America.
authors.forename: Mingzhang
authors.initials: M
authors.lastname: Guo
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Matthew
authors.initials: M
authors.lastname: Rubin
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Jenny
authors.initials: J
authors.lastname: Guido
authors.affiliation: Department of Surgery, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Mack H
authors.initials: MH
authors.lastname: Wu
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine, Tampa, Florida, United States of America.
authors.forename: Sarah Y
authors.initials: SY
authors.lastname: Yuan
mesh_terms: D020413:3' Untranslated Regions; D051722:ADAM Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D015824:Blood-Air Barrier / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D015536:Down-Regulation; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D016130:Immunophenotyping; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D010539:Permeability; D034622:RNA Interference
chemical_list: D020413:3' Untranslated Regions; C572377:MIRN147 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D051722:ADAM Proteins; C098660:ADAM15 protein, human