Akt-mediated transactivation of the s1p1 receptor in caveolin-enriched microdomains regulates endothelial barrier enhancement by oxidized phospholipids

Patrick A. Singleton, Santipongse Chatchavalvanich, Panfeng Fu, Junjie Xing, Anna A. Birukova, Jennifer A. Fortune, Alexander M. Klibanov, Joe GN Garcia, Konstantin G. Birukov

Research output: Contribution to journalArticle

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Abstract

Endothelial cell (EC) barrier dysfunction results in increased vascular permeability, leading to increased mass transport across the vessel wall and leukocyte extravasation, the key mechanisms in pathogenesis of tissue inflammation and edema. We have previously demonstrated that OxPAPC (oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine) significantly enhances vascular endothelial barrier properties in vitro and in vivo and attenuates endothelial hyperpermeability induced by inflammatory and edemagenic agents via Rac and Cdc42 GTPase dependent mechanisms. These findings suggested potential important therapeutic value of barrier-protective oxidized phospholipids. In this study, we examined involvement of signaling complexes associated with caveolin-enriched microdomains (CEMs) in barrier-protective responses of human pulmonary ECs to OxPAPC. Immunoblotting from OxPAPC-treated ECs revealed OxPAPC-mediated rapid recruitment (5 minutes) to CEMs of the sphingosine 1-phosphate receptor (S1P1), the serine/threonine kinase Akt, and the Rac1 guanine nucleotide exchange factor Tiam1 and phosphorylation of caveolin-1, indicative of signaling activation in CEMs. Abolishing CEM formation (methyl-β-cyclodextrin) blocked OxPAPC-mediated Rac1 activation, cytoskeletal reorganization, and EC barrier enhancement. Silencing (small interfering RNA) Akt expression blocked OxPAPC-mediated S1P1 activation (threonine phosphorylation), whereas silencing S1P1 receptor expression blocked OxPAPC-mediated Tiam1 recruitment to CEMs, Rac1 activation, and EC barrier enhancement. To confirm our in vitro results in an in vivo murine model of acute lung injury with pulmonary vascular hyperpermeability, we observed that selective lung silencing of caveolin-1 or S1P1 receptor expression blocked OxPAPC-mediated protection from ventilator-induced lung injury. Taken together, these results suggest Akt-dependent transactivation of S1P1 within CEMs is important for OxPAPC-mediated cortical actin rearrangement and EC barrier protection.

Original languageEnglish (US)
Pages (from-to)978-986
Number of pages9
JournalCirculation Research
Volume104
Issue number8
DOIs
StatePublished - Apr 24 2009
Externally publishedYes

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Lysosphingolipid Receptors
Caveolins
Transcriptional Activation
Phospholipids
Endothelial Cells
Caveolin 1
Lung
Blood Vessels
Ventilator-Induced Lung Injury
Phosphorylation
Guanine Nucleotide Exchange Factors
2-arachidonoylglycero-3-phosphocholine
Cytoprotection
Acute Lung Injury
Protein-Serine-Threonine Kinases
GTP Phosphohydrolases
Cyclodextrins
Capillary Permeability
Threonine
Immunoblotting

Keywords

  • Akt
  • Caveolin-enriched microdomain
  • Endothelial barrier enhancement
  • OxPAPC
  • S1P receptor

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Akt-mediated transactivation of the s1p1 receptor in caveolin-enriched microdomains regulates endothelial barrier enhancement by oxidized phospholipids. / Singleton, Patrick A.; Chatchavalvanich, Santipongse; Fu, Panfeng; Xing, Junjie; Birukova, Anna A.; Fortune, Jennifer A.; Klibanov, Alexander M.; Garcia, Joe GN; Birukov, Konstantin G.

In: Circulation Research, Vol. 104, No. 8, 24.04.2009, p. 978-986.

Research output: Contribution to journalArticle

Singleton, Patrick A. ; Chatchavalvanich, Santipongse ; Fu, Panfeng ; Xing, Junjie ; Birukova, Anna A. ; Fortune, Jennifer A. ; Klibanov, Alexander M. ; Garcia, Joe GN ; Birukov, Konstantin G. / Akt-mediated transactivation of the s1p1 receptor in caveolin-enriched microdomains regulates endothelial barrier enhancement by oxidized phospholipids. In: Circulation Research. 2009 ; Vol. 104, No. 8. pp. 978-986.
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