title: DNA-dependent protein kinase inhibition blocks asthma in mice and modulates human endothelial and CD4⁺ T-cell function without causing severe combined immunodeficiency.
abstract: BACKGROUND
  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.
  
  
  OBJECTIVE
  We sought to investigate the role of DNA-PK in asthma.
  
  
  METHODS
  Cell culture and ovalbumin (OVA)- or house dust mite-based murine asthma models were used in this study.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  Our results suggest DNA-PK as a novel determinant of asthma and a potential target for the treatment of the disease.
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La; Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Ghonim
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La.
authors.forename: Kusma
authors.initials: K
authors.lastname: Pyakurel
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La.
authors.forename: Jihang
authors.initials: J
authors.lastname: Ju
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La.
authors.forename: Paulo C
authors.initials: PC
authors.lastname: Rodriguez
authors.affiliation: Pulmonary and Critical Care Section, LSU Health Sciences Center, New Orleans, La.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Lammi
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La.
authors.forename: Christian
authors.initials: C
authors.lastname: Davis
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La.
authors.forename: Mohammad Q
authors.initials: MQ
authors.lastname: Abughazleh
authors.affiliation: Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.
authors.forename: Moselhy S
authors.initials: MS
authors.lastname: Mansy
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La. Electronic address: anaura@pu.ac.in.
authors.forename: Amarjit S
authors.initials: AS
authors.lastname: Naura
authors.affiliation: Stanley S. Scott Cancer Center, LSU Health Sciences Center, New Orleans, La. Electronic address: hboulr@lsuhsc.edu.
authors.forename: A Hamid
authors.initials: AH
authors.lastname: Boulares
mesh_terms: D019264:Adoptive Transfer; D000485:Allergens / Q000276:immunology; D000818:Animals; D001249:Asthma / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D016535:Bronchial Hyperreactivity / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002448:Cell Adhesion; D016207:Cytokines / Q000378:metabolism; D051747:DNA-Activated Protein Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D004804:Eosinophils / Q000276:immunology / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D050990:GATA3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018740:Genetic Heterogeneity; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009929:Organ Size; D010047:Ovalbumin / Q000009:adverse effects / Q000276:immunology; D010641:Phenotype; D010950:Plasma Cells / Q000276:immunology / Q000378:metabolism; D039981:Pyroglyphidae / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D020545:Respiratory Mucosa / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D016511:Severe Combined Immunodeficiency; D013154:Spleen / Q000033:anatomy & histology / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D018418:Th2 Cells / Q000276:immunology / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism
chemical_list: D000485:Allergens; D016207:Cytokines; D050990:GATA3 Transcription Factor; D011948:Receptors, Antigen, T-Cell; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D007073:Immunoglobulin E; D010047:Ovalbumin; D051747:DNA-Activated Protein Kinase
keywords: DNA-dependent protein kinase; GATA-3; T(H)2 cytokines; T-cell receptor; adhesion molecules; asthma; eosinophilia; severe combined immunodeficiency disease