Experimental Lung Injury Reduces Krüppel-like Factor 2 to Increase Endothelial Permeability via Regulation of RAPGEF3-Rac1 Signaling

Am J Respir Crit Care Med. 2017 Mar 1;195(5):639-651. doi: 10.1164/rccm.201604-0668OC.


Rationale: Acute respiratory distress syndrome (ARDS) is caused by widespread endothelial barrier disruption and uncontrolled cytokine storm. Genome-wide association studies (GWAS) have linked multiple genes to ARDS. Although mechanosensitive transcription factor Krüppel-like factor 2 (KLF2) is a major regulator of endothelial function, its role in regulating pulmonary vascular integrity in lung injury and ARDS-associated GWAS genes remains poorly understood.

Objectives: To examine KLF2 expression in multiple animal models of acute lung injury and further elucidate the KLF2-mediated pathways involved in endothelial barrier disruption and cytokine storm in experimental lung injury.

Methods: Animal and in vitro models of acute lung injury were used to characterize KLF2 expression and its downstream effects responding to influenza A virus (A/WSN/33 [H1N1]), tumor necrosis factor-α, LPS, mechanical stretch/ventilation, or microvascular flow. KLF2 manipulation, permeability measurements, small GTPase activity, luciferase assays, chromatin immunoprecipitation assays, and network analyses were used to determine the mechanistic roles of KLF2 in regulating endothelial monolayer integrity, ARDS-associated GWAS genes, and lung pathophysiology.

Measurements and main results: KLF2 is significantly reduced in several animal models of acute lung injury. Microvascular endothelial KLF2 is significantly induced by capillary flow but reduced by pathologic cyclic stretch and inflammatory stimuli. KLF2 is a novel activator of small GTPase Ras-related C3 botulinum toxin substrate 1 by transcriptionally controlling Rap guanine nucleotide exchange factor 3/exchange factor directly activated by cyclic adenosine monophosphate, which maintains vascular integrity. KLF2 regulates multiple ARDS GWAS genes related to cytokine storm, oxidation, and coagulation in lung microvascular endothelium. KLF2 overexpression ameliorates LPS-induced lung injury in mice.

Conclusions: Disruption of endothelial KLF2 results in dysregulation of lung microvascular homeostasis and contributes to lung pathology in ARDS.

Keywords: GTPase; Rho; acute respiratory distress syndrome; endothelial dysfunction; exchange factor directly activated by cyclic adenosine monophosphate.

MeSH terms

  • Animals
  • Capillary Permeability / physiology*
  • Disease Models, Animal
  • Endothelium, Vascular / metabolism*
  • GTP Phosphohydrolases / metabolism*
  • Guanine Nucleotide Exchange Factors / metabolism
  • Humans
  • Kruppel-Like Transcription Factors / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neuropeptides / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Respiratory Distress Syndrome / metabolism*
  • Signal Transduction / physiology*
  • rac1 GTP-Binding Protein / metabolism


  • Guanine Nucleotide Exchange Factors
  • KLF2 protein, human
  • Klf2 protein, mouse
  • Klf2 protein, rat
  • Kruppel-Like Transcription Factors
  • Neuropeptides
  • RAC1 protein, human
  • RAPGEF3 protein, human
  • Rac1 protein, mouse
  • Rapgef3 protein, rat
  • GTP Phosphohydrolases
  • Rac1 protein, rat
  • rac1 GTP-Binding Protein