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DNA-dependent protein kinase inhibition blocks asthma in mice and modulates human endothelial and CD4+ T-cell function without causing severe combined immunodeficiency

2014; Elsevier BV; Volume: 135; Issue: 2 Linguagem: Inglês

10.1016/j.jaci.2014.09.005

1097-6825

Mohamed A. Ghonim, Kusma Pyakurel, Jihang Ju, Paulo C. Rodrı́guez, Matthew R. Lammi, Christian Davis, Mohammad Q. Abughazleh, Moselhy S. Mansy, Amarjit S. Naura, A. Hamid Boulares,

T-cell and B-cell Immunology

BackgroundWe reported that DNA-dependent protein kinase (DNA-PK) is critical for the expression of nuclear factor κB–dependent genes in TNF-α–treated glioblastoma cells, suggesting an involvement in inflammatory diseases.ObjectiveWe sought to investigate the role of DNA-PK in asthma.MethodsCell culture and ovalbumin (OVA)– or house dust mite–based murine asthma models were used in this study.ResultsDNA-PK was essential for monocyte adhesion to TNF-α–treated endothelial cells. Administration of the DNA-PK inhibitor NU7441 reduced airway eosinophilia, mucus hypersecretion, airway hyperresponsiveness, and OVA-specific IgE production in mice prechallenged with OVA. Such effects correlated with a marked reduction in lung vascular cell adhesion molecule 1 expression and production of several cytokines, including IL-4, IL-5, IL-13, eotaxin, IL-2, and IL-12 and the chemokines monocyte chemoattractant protein 1 and keratinocyte-derived chemokine, with a negligible effect on IL-10/IFN-γ production. DNA-PK inhibition by gene heterozygosity of the 450-kDa catalytic subunit of the kinase (DNA-PKcs+/−) also prevented manifestation of asthma-like traits. These results were confirmed in a chronic model of asthma by using house dust mite, a human allergen. Remarkably, such protection occurred without causing severe combined immunodeficiency. Adoptive transfer of TH2-skewed OT-II wild-type CD4+ T cells reversed IgE and TH2 cytokine production but not airway hyperresponsiveness in OVA-challenged DNA-PKcs+/− mice. DNA-PK inhibition reduced IL-4, IL-5, IL-13, eotaxin, IL-8, and monocyte chemoattractant protein 1 production without affecting IL-2, IL-12, IFN-γ, and interferon-inducible protein 10 production in CD3/CD28-stimulated human CD4+ T cells, potentially by blocking expression of Gata3. These effects occurred without significant reductions in T-cell proliferation. In mouse CD4+ T cells in vitro DNA-PK inhibition severely blocked CD3/CD28-induced Gata3 and T-bet expression in CD4+ T cells and prevented differentiation of TH1 and TH2 cells under respective TH1- and TH2-skewing conditions.ConclusionOur results suggest DNA-PK as a novel determinant of asthma and a potential target for the treatment of the disease. We reported that DNA-dependent protein kinase (DNA-PK) is critical for the expression of nuclear factor κB–dependent genes in TNF-α–treated glioblastoma cells, suggesting an involvement in inflammatory diseases. We sought to investigate the role of DNA-PK in asthma. Cell culture and ovalbumin (OVA)– or house dust mite–based murine asthma models were used in this study. DNA-PK was essential for monocyte adhesion to TNF-α–treated endothelial cells. Administration of the DNA-PK inhibitor NU7441 reduced airway eosinophilia, mucus hypersecretion, airway hyperresponsiveness, and OVA-specific IgE production in mice prechallenged with OVA. Such effects correlated with a marked reduction in lung vascular cell adhesion molecule 1 expression and production of several cytokines, including IL-4, IL-5, IL-13, eotaxin, IL-2, and IL-12 and the chemokines monocyte chemoattractant protein 1 and keratinocyte-derived chemokine, with a negligible effect on IL-10/IFN-γ production. DNA-PK inhibition by gene heterozygosity of the 450-kDa catalytic subunit of the kinase (DNA-PKcs+/−) also prevented manifestation of asthma-like traits. These results were confirmed in a chronic model of asthma by using house dust mite, a human allergen. Remarkably, such protection occurred without causing severe combined immunodeficiency. Adoptive transfer of TH2-skewed OT-II wild-type CD4+ T cells reversed IgE and TH2 cytokine production but not airway hyperresponsiveness in OVA-challenged DNA-PKcs+/− mice. DNA-PK inhibition reduced IL-4, IL-5, IL-13, eotaxin, IL-8, and monocyte chemoattractant protein 1 production without affecting IL-2, IL-12, IFN-γ, and interferon-inducible protein 10 production in CD3/CD28-stimulated human CD4+ T cells, potentially by blocking expression of Gata3. These effects occurred without significant reductions in T-cell proliferation. In mouse CD4+ T cells in vitro DNA-PK inhibition severely blocked CD3/CD28-induced Gata3 and T-bet expression in CD4+ T cells and prevented differentiation of TH1 and TH2 cells under respective TH1- and TH2-skewing conditions. Our results suggest DNA-PK as a novel determinant of asthma and a potential target for the treatment of the disease.