Combinatorial Therapy with Acetylation and Methylation Modifiers Attenuates Lung Vascular Hyperpermeability in Endotoxemia-Induced Mouse Inflammatory Lung Injury

Impairment of tissue fluid homeostasis and migration of inflammatory cells across the vascular endothelial barrier are crucial factors in the pathogenesis of acute lung injury (ALI). The goal for treatment of ALI is to target pathways that lead to profound dysregulation of the lung endothelial barrier. Although studies have shown that chemical epigenetic modifiers can limit lung inflammation in experimental ALI models, studies to date have not examined efficacy of a combination of DNA methyl transferase inhibitor 5-Aza 2-deoxycytidine and histone deacetylase inhibitor trichostatin A (herein referred to as Aza+TSA) after endotoxemia-induced mouse lung injury. We tested the hypothesis that treatment with Aza+TSA after lipopolysaccharide induction of ALI through epigenetic modification of lung endothelial cells prevents inflammatory lung injury. Combinatorial treatment with Aza+TSA mitigated the increased endothelial permeability response after lipopolysaccharide challenge. In addition, we observed reduced lung inflammation and lung injury. Aza+TSA also significantly reduced mortality in the ALI model. The protection was ascribed to inhibition of the eNOS-Cav1-MLC2 signaling pathway and enhanced acetylation of histone markers on the vascular endothelial-cadherin promoter. In summary, these data show for the first time the efficacy of combinatorial Aza+TSA therapy in preventing ALI in lipopolysaccharide-induced endotoxemia and raise the possibility of an essential role of DNA methyl transferase and histone deacetylase in the mechanism of ALI. Impairment of tissue fluid homeostasis and migration of inflammatory cells across the vascular endothelial barrier are crucial factors in the pathogenesis of acute lung injury (ALI). The goal for treatment of ALI is to target pathways that lead to profound dysregulation of the lung endothelial barrier. Although studies have shown that chemical epigenetic modifiers can limit lung inflammation in experimental ALI models, studies to date have not examined efficacy of a combination of DNA methyl transferase inhibitor 5-Aza 2-deoxycytidine and histone deacetylase inhibitor trichostatin A (herein referred to as Aza+TSA) after endotoxemia-induced mouse lung injury. We tested the hypothesis that treatment with Aza+TSA after lipopolysaccharide induction of ALI through epigenetic modification of lung endothelial cells prevents inflammatory lung injury. Combinatorial treatment with Aza+TSA mitigated the increased endothelial permeability response after lipopolysaccharide challenge. In addition, we observed reduced lung inflammation and lung injury. Aza+TSA also significantly reduced mortality in the ALI model. The protection was ascribed to inhibition of the eNOS-Cav1-MLC2 signaling pathway and enhanced acetylation of histone markers on the vascular endothelial-cadherin promoter. In summary, these data show for the first time the efficacy of combinatorial Aza+TSA therapy in preventing ALI in lipopolysaccharide-induced endotoxemia and raise the possibility of an essential role of DNA methyl transferase and histone deacetylase in the mechanism of ALI. Sepsis-induced acute lung injury (ALI) is an inflammatory disorder that affects 18 million people worldwide and 200,000 people in the United States each year1Nemeth K. Mayer B. Mezey E. Modulation of bone marrow stromal cell functions in infectious diseases by toll-like receptor ligands.J Mol Med (Berl). 2010; 88: 5-10Crossref PubMed Scopus (64) Google Scholar, 2Ware L.B. Matthay M.A. The acute respiratory distress syndrome.N Engl J Med. 2000; 342: 1334-1349Crossref PubMed Scopus (4529) Google Scholar and is associated with a mortality rate of approximately 30% despite advanced interventions.3Price L.C. McAuley D.F. Marino P.S. Finney S.J. Griffiths M.J. Wort S.J. 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PubMed Scopus Google Scholar in endothelial cells of and in with increased increased and of J. signaling and endothelial Cell PubMed Scopus Google Scholar This a and disruption of the vascular endothelial J. signaling and endothelial Cell PubMed Scopus Google Scholar data show that lung endothelial cells to a in was by treatment with in the mouse lung that significantly increased lung vascular in was also significantly reduced in with We observed a in expression of in lung endothelial cells to was with Aza+TSA The in was and Aza+TSA treatment to the of these epigenetic the of and endothelial modifiers epigenetic with that are The lead to of DNA not in DNA T. Allis C.D. Translating the histone code.Science. 2001; 293: 1074-1080Crossref PubMed Scopus (7709) Google Scholar, P. and their on Full Text Full Text PDF PubMed Scopus Google Scholar inflammation was shown to be associated with the acetylation of in and B. F. M. D.R. cell of histone deacetylase inhibition in Full Text Full Text PDF PubMed Scopus Google Scholar we the possibility that gene expression in endothelial cells after is by histone studies increased of histone and histone and reduced acetylation of histone 18 and histone are of and P. P. Wang C.J. C.Y. and cell gene to the in Cell PubMed Scopus Google Scholar We examined the of these histone markers in the of the reduced acetylation of histone and in in endothelial and treatment of cells with Aza+TSA increased we observed reduced methylation of histone in in endothelial was increased on treatment with data that the endothelial barrier effect of with combination of Aza+TSA is the of acetylation and methylation of histone the of promoter. studies after of a significantly survival rate in in of and of inflammatory in lung also significantly reduced in with Aza mortality to the with as as the first of the potential of In a of Aza+TSA after of ALI lung vascular and inflammatory lung injury. 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