TLR4 activation induces inflammatory vascular permeability via Dock1 targeting and NOX4 upregulation

Jin H. Song, Joseph B. Mascarenhas, Saad Sammani, Carrie L. Kempf, Hua Cai, Sara M. Camp, Tadeo Bermudez, Donna D. Zhang, Viswanathan Natarajan, Joe G.N. Garcia

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The loss of vascular integrity is a cardinal feature of acute inflammatory responses evoked by activation of the TLR4 inflammatory cascade. Utilizing in vitro and in vivo models of inflammatory lung injury, we explored TLR4-mediated dysregulated signaling that results in the loss of endothelial cell (EC) barrier integrity and vascular permeability, focusing on Dock1 and Elmo1 complexes that are intimately involved in regulation of Rac1 GTPase activity, a well recognized modulator of vascular integrity. Marked reductions in Dock1 and Elmo1 expression was observed in lung tissues (porcine, rat, mouse) exposed to TLR4 ligand-mediated acute inflammatory lung injury (LPS, eNAMPT) in combination with injurious mechanical ventilation. Lung tissue levels of Dock1 and Elmo1 were preserved in animals receiving an eNAMPT-neutralizing mAb in conjunction with highly significant decreases in alveolar edema and lung injury severity, consistent with Dock1/Elmo1 as pathologic TLR4 targets directly involved in inflammation-mediated loss of vascular barrier integrity. In vitro studies determined that pharmacologic inhibition of Dock1-mediated activation of Rac1 (TBOPP) significantly exacerbated TLR4 agonist-induced EC barrier dysfunction (LPS, eNAMPT) and attenuated increases in EC barrier integrity elicited by barrier-enhancing ligands of the S1P1 receptor (sphingosine-1-phosphate, Tysiponate). The EC barrier-disrupting influence of Dock1 inhibition on S1PR1 barrier regulation occurred in concert with: 1) suppressed formation of EC barrier-enhancing lamellipodia, 2) altered nmMLCK-mediated MLC2 phosphorylation, and 3) upregulation of NOX4 expression and increased ROS. These studies indicate that Dock1 is essential for maintaining EC junctional integrity and is a critical target in TLR4-mediated inflammatory lung injury.

Original languageEnglish (US)
Article number166562
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1868
Issue number12
DOIs
StatePublished - Dec 1 2022

Keywords

  • Dock1
  • EC barrier integrity
  • Elmo1
  • Oxidative stress
  • Vascular permeability
  • eNAMPT

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology

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