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Download fileIn-Depth Proteomics Identifies a Role for Autophagy in Controlling Reactive Oxygen Species Mediated Endothelial Permeability
journal contribution
posted on 01.06.2016, 00:00 authored by Francesca Patella, Lisa J. Neilson, Dimitris Athineos, Zahra Erami, Kurt I. Anderson, Karen Blyth, Kevin
M. Ryan, Sara ZanivanEndothelial
cells (ECs) form the inner layer of blood vessels and
physically separate the blood from the surrounding tissue. To support
tissues with nutrients and oxygen, the endothelial monolayer is semipermeable.
When EC permeability is altered, blood vessels are not functional,
and this is associated with disease. A comprehensive knowledge of
the mechanisms regulating EC permeability is key in developing strategies
to target this mechanism in pathologies. Here we have used an in vitro
model of human umbilical vein endothelial cells mimicking the formation
of a physiologically permeable vessel and performed time-resolved
in-depth molecular profiling using stable isotope labeling by amino
acids in cell culture mass spectrometry (MS)-proteomics. Autophagy
is induced when ECs are assembled into a physiologically permeable
monolayer. By using siRNA and drug treatment to block autophagy in
combination with functional assays and MS proteomics, we show that
ECs require autophagy flux to maintain intracellular reactive oxygen
species levels, and this is required to maintain the physiological
permeability of the cells.