title: Slit2-Robo4 receptor responses inhibit ANDV directed permeability of human lung microvascular endothelial cells.
abstract: Hantaviruses nonlytically infect human endothelial cells (ECs) and cause edematous and hemorrhagic diseases. Andes virus (ANDV) causes hantavirus pulmonary syndrome (HPS), and Hantaan virus (HTNV) causes hemorrhagic fever with renal syndrome (HFRS). Hantaviruses enhance vascular endothelial growth factor directed EC permeability resulting in the disassembly of inter-endothelial cell adherens junctions (AJs). Recent studies demonstrate that Slit2 binding to Robo1/Robo4 receptors on ECs has opposing effects on AJ disassembly and vascular fluid barrier functions. Here we demonstrate that Slit2 inhibits ANDV and HTNV induced permeability and AJ disassembly of pulmonary microvascular ECs (PMECs) by interactions with Robo4. In contrast, Slit2 had no effect on the permeability of ANDV infected human umbilical vein ECs (HUVECs). Analysis of Robo1/Robo4 expression determined that PMECs express Robo4, but not Robo1, while HUVECs expressed both Robo4 and Robo1 receptors. SiRNA knockdown of Robo4 in PMECs prevented Slit2 inhibition of ANDV induced permeability demonstrating that Robo4 receptors determine PMEC responsiveness to Slit2. Collectively, this data demonstrates a selective role for Slit2/Robo4 responses within PMECs that inhibits ANDV induced permeability and AJ disassembly. These findings suggest Slit2s utility as a potential HPS therapeutic that stabilizes the pulmonary endothelium and antagonizes ANDV induced pulmonary edema.
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794-512, United States.
authors.forename: Elena E
authors.initials: EE
authors.lastname: Gorbunova
authors.forename: Irina N
authors.initials: IN
authors.lastname: Gavrilovskaya
authors.forename: Erich R
authors.initials: ER
authors.lastname: Mackow
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism / Q000648:ultrastructure; D002199:Capillary Permeability; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000502:physiology; D018063:Hantaan virus / Q000502:physiology; D006476:Orthohantavirus / Q000502:physiology*; D018804:Hantavirus Pulmonary Syndrome / Q000821:virology*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000821:virology; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology* / Q000821:virology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D008168:Lung / Q000098:blood supply* / Q000378:metabolism; D055806:Microvessels / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000378:metabolism; D000097905:Roundabout Proteins; D000098552:Slit Homolog 2 Protein
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D000097905:Roundabout Proteins; D000098552:Slit Homolog 2 Protein; C462951:ROBO4 protein, human
keywords: Hantavirus; Human endothelial cells; Inhibition; Permeability; Slit2