p18, a novel adaptor protein, regulates pulmonary endothelial barrier function via enhanced endocytic recycling of VE-cadherin

FASEB J. 2015 Mar;29(3):868-81. doi: 10.1096/fj.14-257212. Epub 2014 Nov 17.

Abstract

Vascular permeability is a hallmark of several disease states including acute lung injury (ALI). Endocytosis of VE-cadherin, away from the interendothelial junction (IEJ), causes acute endothelial barrier permeability. A novel protein, p18, anchors to the endosome membrane and plays a role in late endosomal signaling via MAPK and mammalian target of rapamycin. However, the fate of the VE-cadherin-positive endosome has yet to be elucidated. We sought to elucidate a role for p18 in VE-cadherin trafficking and thus endothelial barrier function, in settings of ALI. Endothelial cell (EC) resistance, whole-cell ELISA, and filtration coefficient were studied in mice or lung ECs overexpressing wild-type or nonendosomal-binding mutant p18, using green fluorescent protein as a control. We demonstrate a protective role for the endocytic protein p18 in endothelial barrier function in settings of ALI in vitro and in vivo, through enhanced recycling of VE-cadherin-positive early endosomes to the IEJ. In settings of LPS-induced ALI, we show that Src tethered to the endosome tyrosine phosphorylates p18 concomitantly with VE-cadherin internalization and pulmonary edema formation. We conclude that p18 regulates pulmonary endothelial barrier function in vitro and in vivo, by enhancing recycling of VE-cadherin-positive endosomes to the IEJ.

Keywords: adherensjunction; endocytosis; endosome; lunginjury.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Lung Injury / chemically induced
  • Acute Lung Injury / metabolism
  • Acute Lung Injury / pathology*
  • Adherens Junctions
  • Animals
  • Antigens, CD / metabolism*
  • Blotting, Western
  • Cadherins / metabolism*
  • Capillary Permeability / drug effects*
  • Cells, Cultured
  • Endocytosis / drug effects*
  • Endosomes / drug effects
  • Endosomes / metabolism
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology*
  • Enzyme-Linked Immunosorbent Assay
  • Immunoenzyme Techniques
  • Immunoprecipitation
  • Lipopolysaccharides / toxicity
  • Mice
  • Mice, Inbred C57BL
  • NM23 Nucleoside Diphosphate Kinases / metabolism*
  • Phosphorylation / drug effects
  • Pulmonary Edema / chemically induced
  • Pulmonary Edema / metabolism
  • Pulmonary Edema / pathology*
  • Signal Transduction
  • Tyrosine / metabolism
  • src-Family Kinases / metabolism

Substances

  • Antigens, CD
  • Cadherins
  • Lipopolysaccharides
  • NM23 Nucleoside Diphosphate Kinases
  • cadherin 5
  • Tyrosine
  • src-Family Kinases
  • Nme2 protein, mouse