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.

CONCLUSION:

Our results suggest DNA-PK as a novel determinant of asthma and a potential target for the treatment of the disease.

• Authors: Mohamed A Ghonim, Kusma Pyakurel, Jihang Ju, Paulo C Rodriguez, Matthew R Lammi, Christian Davis, Mohammad Q Abughazleh, Moselhy S Mansy, Amarjit S Naura, A Hamid Boulares
• Journal: The Journal of allergy and clinical immunology (J Allergy Clin Immunol) Vol. 135 Issue 2 Pg. 425-40 (Feb 2015) ISSN: 1097-6825 [Electronic] United States
• PMID: 25441643 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2014 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Chemical References

• Allergens
• Cytokines
• GATA3 Transcription Factor
• Receptors, Antigen, T-Cell
• Tumor Necrosis Factor-alpha
• Vascular Cell Adhesion Molecule-1
• Immunoglobulin E
• Ovalbumin
• DNA-Activated Protein Kinase

Topics

• Adoptive Transfer
• Allergens (immunology)
• Animals
• Asthma (immunology, metabolism, pathology)
• Bronchial Hyperreactivity (immunology, metabolism, pathology)
• CD4-Positive T-Lymphocytes (immunology)
• Cell Adhesion
• Cytokines (metabolism)
• DNA-Activated Protein Kinase (antagonists & inhibitors, genetics, metabolism)
• Disease Models, Animal
• Eosinophils (immunology, metabolism)
• Epithelial Cells (metabolism)
• GATA3 Transcription Factor (genetics, metabolism)
• Gene Expression
• Genetic Heterogeneity
• Humans
• Immunoglobulin E (immunology)
• Lymphocyte Activation
• Male
• Mice
• Mice, Knockout
• Organ Size
• Ovalbumin (adverse effects, immunology)
• Phenotype
• Plasma Cells (immunology, metabolism)
• Pyroglyphidae (immunology)
• Receptors, Antigen, T-Cell (metabolism)
• Respiratory Mucosa (immunology, metabolism, pathology)
• Severe Combined Immunodeficiency
• Spleen (anatomy & histology, immunology)
• T-Lymphocyte Subsets (immunology, metabolism)
• Th2 Cells (immunology, metabolism)
• Tumor Necrosis Factor-alpha (metabolism)
• Vascular Cell Adhesion Molecule-1 (genetics, metabolism)