Hepatocyte growth factor enhances endothelial cell barrier function and cortical cytoskeletal rearrangement: potential role of glycogen synthase kinase‐3β

Liu, Feng; Schaphorst, Kane L.; Verin, Alexander D.; Jacobs, Keri; Birukova, Anna A.; Day, Regina M.; Bogatcheva, Natalia V.; Bottaro, Donald P.; Garcia, Joe G.N.

doi:10.1096/fj.01-0870com

Cited by 157 publications

(176 citation statements)

References 64 publications

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“…EC barrier enhancement induced by oxidized phospholipids, sphingosine 1-phosphate, hepatocyte growth factor [10,13,47] or prostaglandins (this study) is tightly associated with actomyosin remodeling characterized by Rac-dependent formation of sub-cortical F-actin rim and disappearance of central stress fibers. In the next experiments we explored a role of small GTPases Rap1 and Rac in PGE 2 -induced actin cytoskeletal remodeling using transfection of pulmonary EC with specific siRNAs.…”

Section: Role Of Rap1 and Rac In Pg-induced Cytoskeletal And Adherensmentioning

confidence: 86%

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Prostaglandins PGE2 and PGI2 promote endothelial barrier enhancement via PKA- and Epac1/Rap1-dependent Rac activation

Birukova

Zagranichnaya

et al. 2007

Experimental Cell Research

262

267

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Prostaglandin E 2 (PGE 2 ) and prostacyclin are lipid mediators produced by cyclooxygenase and implicated in the regulation of vascular function, wound repair, inflammatory processes, and acute lung injury. Although protective effects of these prostaglandins (PGs) are associated with stimulation of intracellular cAMP production, the crosstalk between cAMP-activated signal pathways in the regulation of endothelial cell (EC) permeability is not well understood. We studied involvement of cAMP-dependent kinase (PKA), cAMP-Epac-Rap1 pathway, and small GTPase Rac in the PGsinduced EC barrier protective effects and cytoskeletal remodeling. PGE 2 and PGI 2 synthetic analog beraprost increased transendothelial electrical resistance and decreased dextran permeability, enhanced peripheral F-actin rim and increased intercellular adherens junction areas reflecting EC barrier-protective response. Furthermore, beraprost dramatically attenuated thrombin-induced Rho activation, MLC phosphorylation and EC barrier dysfunction. In vivo, beraprost attenuated lung barrier dysfunction induced by high tidal volume mechanical ventilation. Both PGs caused cAMPmediated activation of PKA-, Epac/Rap1-and Tiam1/Vav2-dependent pathways of Rac1 activation and EC barrier regulation. Knockdown of Epac, Rap1, Rac-specific exchange factors Tiam1 and Vav2 using siRNA approach, or inhibition of PKA activity decreased Rac1 activation and PGinduced EC barrier enhancement. Thus, our results show that barrier-protective effects of PGE 2 and prostacyclin on pulmonary EC are mediated by PKA and Epac/Rap pathways, which converge on Rac activation and lead to enhancement of peripheral actin cytoskeleton and adherens junctions. These mechanisms may mediate protective effects of PGs against agonist-induced lung vascular barrier dysfunction in vitro and against mechanical stress-induced lung injury in vivo.

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Section: Role Of Rap1 and Rac In Pg-induced Cytoskeletal And Adherensmentioning

confidence: 86%

“…We and others have previously described mechanisms of Rac activation by various groups of barrier-protective agonists such as sphingosine 1-phosphate, HGF and oxidized phosphocholine and characterized a role of Rac in EC cytoskeletal remodeling and pulmonary EC barrier protective responses [10,11,13,47,48].…”

Section: Discussionmentioning

confidence: 99%

Prostaglandins PGE2 and PGI2 promote endothelial barrier enhancement via PKA- and Epac1/Rap1-dependent Rac activation

Birukova

Zagranichnaya

et al. 2007

Experimental Cell Research

262

267

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“…Activation of small GTPase Rac has been previously described as a leading mechanism of barrier protection in response to various stimuli including OxPAPC, sphingosine 1-phosphate, hepatocyte growth factor (HGF), and shear stress (Garcia, Liu et al 2001;Birukov, Birukova et al 2002;Liu, Schaphorst et al 2002;Birukov, Bochkov et al 2004;Shikata, Rios et al 2005). In the next studies we investigated the roles of protein kinases A and C and tyrosine kinases in the regulation of OxPAPC-induced Rac activation.…”

Section: Involvement Of Pka and Pkc-dependent Pathways In Oxpapc-indumentioning

confidence: 94%

“…A number of agonists including phospholipid products (Zhang, Magnusson et al 1997;Huang, Subbaiah et al 2005) and bioactive peptides (Garcia, Siflinger-Birnboim et al 1986;van Nieuw Amerongen, Draijer et al 1998;Petrache, Verin et al 2001;Moy, Blackwell et al 2002;Birukova, Adyshev et al 2005) increase vascular endothelial permeability, which my lead to alveolar flooding and pulmonary edema. However, much smaller number of bioactive molecules capable of enhancing endothelial barrier properties have been reported so far (Garcia, Liu et al 2001;Liu, Schaphorst et al 2002;VouretCraviari, Bourcier et al 2002;Birukov, Bochkov et al 2004;Kolosova, Mirzapoiazova et al 2005), and signaling mechanisms underlying these barrier protective effects have been a major focus of ongoing studies (Lee, Lee et al 2000;Garcia, Liu et al 2001;Vouret-Craviari, Bourcier et al 2002;Birukov, Leitinger et al 2004;Dudek, Jacobson et al 2004;Kolosova, Mirzapoiazova et al 2005;Mehta, Konstantoulaki et al 2005;Singleton, Dudek et al 2005). …”

Section: Introductionmentioning

confidence: 99%

Signaling pathways involved in OxPAPC-induced pulmonary endothelial barrier protection

Birukova

Chatchavalvanich

Oskolkova

et al. 2007

Microvascular Research

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Increased tissue or serum levels of oxidized phospholipids have been detected in a variety of chronic and acute pathological conditions such as hyperlipidemia, atherosclerosis, heart attack, cell apoptosis, acute inflammation and injury. We have recently described signaling cascades activated by oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (OxPAPC) in the human pulmonary artery endothelial cells (EC) and reported potent barrier-protective effects of OxPAPC, which were mediated by small GTPases Rac and Cdc42. In this study we have further characterized signal transduction pathways involved in the OxPAPC-mediated endothelial barrier protection. Inhibitors of small GTPases, protein kinase A (PKA), protein kinase C (PKC), Src family kinases and general inhibitors of tyrosine kinases attenuated OxPAPC-induced barrier-protective response and EC cytoskeletal remodeling. In contrast, small GTPase Rho, Rho kinase, Erk-1,2 MAP kinase and p38 MAP kinase and PI3-kinase were not involved in the barrier-protective effects of OxPAPC. Inhibitors of PKA, PKC, tyrosine kinases and small GTPase inhibitor toxin B suppressed OxPAPC-induced Rac activation and decreased phosphorylation of focal adhesion kinase (FAK) and paxillin. Barrierprotective effects of OxPAPC were not reproduced by platelet activating factor (PAF), which at high concentrations induced barrier dysfunction, but were partially attenuated by PAF receptor antagonist A85783. These results demonstrate for the first time upstream signaling cascades involved in the OxPAPC-induced Rac activation, cytoskeletal remodeling and barrier regulation and suggest PAF receptor-independent mechanisms of OxPAPC-mediated endothelial barrier protection.

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“…Akt is involved in a number of cell processes including cell growth, proliferation, survival (23); angiogenesis; vascular maturation; regulation of EC permeability; and EC nitric oxide synthase activity (24,25). We previously demonstrated the direct contribution of Akt to hepatocyte growth factor-mediated enhancement of lung vascular barrier properties during lung inflammation via GSK3-B activation (26). Evidence also supports an important role for Akt in the pathogenesis of VILI because pharmacologic inhibition of Akt results in augmented microvascular permeability in overdistended isolated mouse lungs (11).…”

Section: Discussionmentioning

confidence: 99%

Role of Growth Arrest and DNA Damage–inducible α in Akt Phosphorylation and Ubiquitination after Mechanical Stress-induced Vascular Injury

Mitra

Sammani

Wang

et al. 2011

Am J Respir Crit Care Med

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Rationale: The stress-induced growth arrest and DNA damageinducible a (GADD45a) gene is up-regulated by mechanical stress with GADD45a knockout (GADD45a 2/2 ) mice demonstrating both increased susceptibility to ventilator-induced lung injury (VILI) and reduced levels of the cell survival and vascular permeability signaling effector (Akt). However, the functional role of GADD45a in the pathogenesis of VILI is unknown. Objectives: We sought to define the role of GADD45a in the regulation of Akt activation induced by mechanical stress. Methods: VILI-challenged GADD45a 2/2 mice were administered a constitutively active Akt1 vector and injury was assessed by bronchoalveolar lavage cell counts and protein levels. Human pulmonary artery endothelial cells (EC) were exposed to 18% cyclic stretch (CS) under conditions of GADD45a silencing and used for immunoprecipitation, Western blotting or immunofluoresence. EC were also transfected with mutant ubiquitin vectors to characterize site-specific Akt ubiquitination. DNA methylation was measured using methylspecific polymerase chain reaction assay. Measurements and Main Results: Studies exploring the linkage of GADD45a with mechanical stress and Akt regulation revealed VILIchallenged GADD45a 2/2 mice to have significantly reduced lung injury on overexpression of Akt1 transgene. Increased mechanical stress with 18% CS in EC induced Akt phosphorylation via E3 ligase tumor necrosis factor receptor-associated factor 6 (TRAF6)-mediated Akt K63 ubiquitination resulting in Akt trafficking and activation at the membrane. GADD45a is essential to this process because GADD45a-silenced endothelial cells and GADD45a 2/2 mice exhibited increased Akt K48 ubiquitination leading to proteasomal degradation. These events involve loss of ubiquitin carboxyl terminal hydrolase 1 (UCHL1), a deubiquitinating enzyme that normally removes K48 polyubiquitin chains bound to Akt thus promoting Akt K63 ubiquitination. Loss of GADD45a significantly reduces UCHL1 expression via UCHL1 promoter methylation resulting in increased Akt K48 ubiquitination and reduced Akt levels. Conclusions: These studies highlight a novel role for GADD45a in the regulation of site-specific Akt ubiquitination and activation and implicate a significant functional role for GADD45a in the clinical predisposition to VILI.Keywords: GADD45a; AKT; UCHL1; ubiquitin; mechanical stress Acute lung injury (ALI) is a devastating disorder commonly encountered in the intensive care unit, with approximately 75,000 deaths annually in the United States (1). Although insults, such as infection and trauma, often precipitate in ALI, ALI morbidity and mortality are heavily influenced by excessive mechanical stress produced by mechanical ventilation. Mechanical ventilator support is required to provide adequate gas exchange but potentially generates excessive mechanical stress, a condition known as ventilator-induced lung injury (VILI) (2). Multiple lines of evidence, including marked ethnic and racial disparities in ALI mortality, support the contribu...

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Hepatocyte growth factor enhances endothelial cell barrier function and cortical cytoskeletal rearrangement: potential role of glycogen synthase kinase‐3β

Cited by 157 publications

References 64 publications

Prostaglandins PGE2 and PGI2 promote endothelial barrier enhancement via PKA- and Epac1/Rap1-dependent Rac activation

Prostaglandins PGE2 and PGI2 promote endothelial barrier enhancement via PKA- and Epac1/Rap1-dependent Rac activation

Signaling pathways involved in OxPAPC-induced pulmonary endothelial barrier protection

Role of Growth Arrest and DNA Damage–inducible α in Akt Phosphorylation and Ubiquitination after Mechanical Stress-induced Vascular Injury

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