UCHL1, a deubiquitinating enzyme, regulates lung endothelial cell permeability in vitro and in vivo

Am J Physiol Lung Cell Mol Physiol. 2021 Apr 1;320(4):L497-L507. doi: 10.1152/ajplung.00492.2020. Epub 2021 Jan 13.

Abstract

Increasing evidence suggests an important role for deubiquitinating enzymes (DUBs) in modulating a variety of biological functions and diseases. We previously identified the upregulation of the DUB ubiquitin carboxyl terminal hydrolase 1 (UCHL1) in murine ventilator-induced lung injury (VILI). However, the role of UCHL1 in modulating vascular permeability, a cardinal feature of acute lung injury (ALI) in general, remains unclear. We investigated the role of UCHL1 in pulmonary endothelial cell (EC) barrier function in vitro and in vivo and examined the effects of UCHL1 on VE-cadherin and claudin-5 regulation, important adherens and tight junctional components, respectively. Measurements of transendothelial electrical resistance confirmed decreased barrier enhancement induced by hepatocyte growth factor (HGF) and increased thrombin-induced permeability in both UCHL1-silenced ECs and in ECs pretreated with LDN-57444 (LDN), a pharmacological UCHL1 inhibitor. In addition, UCHL1 knockdown (siRNA) was associated with decreased expression of VE-cadherin and claudin-5, whereas silencing of the transcription factor FoxO1 restored claudin-5 levels. Finally, UCHL1 inhibition in vivo via LDN was associated with increased VILI in a murine model. These findings support a prominent functional role of UCHL1 in regulating lung vascular permeability via alterations in adherens and tight junctions and implicate UCHL1 as an important mediator of ALI.

Keywords: UCHL1; VE-cadherin; adherens junction; claudin-5; endothelial cells; lung vascular permeability; tight junction.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Capillary Permeability*
  • Cells, Cultured
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology*
  • In Vitro Techniques
  • Indoles / pharmacology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Oximes / pharmacology
  • Signal Transduction
  • Ubiquitin Thiolesterase / antagonists & inhibitors
  • Ubiquitin Thiolesterase / genetics
  • Ubiquitin Thiolesterase / metabolism*
  • Ubiquitination
  • Ventilator-Induced Lung Injury / metabolism
  • Ventilator-Induced Lung Injury / pathology*

Substances

  • Indoles
  • LDN 57444
  • Oximes
  • Ubiquitin Thiolesterase
  • Uchl1 protein, mouse