Modulation by aspirin of nuclear phospho–signal transducer and activator of transcription 6 expression: Possible role in therapeutic benefit associated with aspirin desensitization

• Published in: Journal of allergy and clinical immunology Vol. 124; no. 4; pp. 724 - 730.e4
• Main Authors: Steinke, John W., PhD, Culp, Jeffrey A., MD, Kropf, Elizabeth, BS, Borish, Larry, MD
• Format: Journal Article
• Language: English
• Published: New York, NY Mosby, Inc 01.10.2009; Elsevier; Elsevier Limited
• Subjects: Allergy and Immunology; Anti-Inflammatory Agents, Non-Steroidal - adverse effects; Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Aspirin; Aspirin - adverse effects; Aspirin - pharmacology; Aspirin-exacerbated respiratory disease; Asthma; Asthma - immunology; Binding sites; Biological and medical sciences; Cell Line; Cyclooxygenase 2 - drug effects; Cyclooxygenase 2 - metabolism; cysteinyl leukotrienes; Cytokines; Desensitization, Immunologic; Drug Hypersensitivity - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Gene expression; Gene Expression - drug effects; Gene Expression - physiology; Glutathione Transferase; Humans; IL-4; Immunopathology; Interleukin-4 - pharmacology; Intramolecular Oxidoreductases - antagonists & inhibitors; Intramolecular Oxidoreductases - metabolism; Ketorolac - pharmacology; Kinases; Medical sciences; Monocytes - drug effects; Monocytes - immunology; Monocytes - metabolism; Nonsteroidal anti-inflammatory drugs; Prostaglandin-E Synthases; Receptors, Leukotriene - drug effects; Receptors, Leukotriene - metabolism; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; signal transduction; Signal Transduction - drug effects; Signal Transduction - physiology; Sodium Salicylate - pharmacology; STAT6 Transcription Factor - antagonists & inhibitors; STAT6 Transcription Factor - genetics; STAT6 Transcription Factor - metabolism; Studies; Up-Regulation - drug effects; Up-Regulation - physiology; Cysteinyl leukotriene; Aspirin-exacerbated respiratory disease; NSAID; AERD; LT; LTC 4S; CysLT; Cysteinyl leukotriene type 1 receptor; Phospho–signal transducer and activator of transcription 6; NF-κB; CysLT1R; Signal transducer and activator of transcription 6; EMSA; Electrophoretic mobility shift assay; STAT6; Nuclear factor κB; mPGES-1; signal transduction; pSTAT6; IL-4; Microsomal prostaglandin E synthase 1; Leukotriene; Leukotriene C 4 synthase; Nonsteroidal anti-inflammatory drug; cysteinyl leukotrienes; Prostaglandin-endoperoxide synthase; Immunopathology; Desensitization; Enzyme; Respiratory disease; Arachidonic acid derivatives; Cytokine; Enzyme inhibitor; Acetylsalicylic acid; Antiplatelet agent; Signal transduction; Interleukin 4; Association; Immunology; Treatment; Transcription factor STAT6; Eicosanoid; Modulation; Oxidoreductases; Salicylates
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2009.07.031

Background Aspirin-exacerbated respiratory disease is characterized by asthma, nasal polyps, and intolerance to aspirin with overexpression of leukotriene (LT) C4 synthase and cysteinyl leukotriene receptors. Through an unknown mechanism, aspirin desensitization is an effective treatment. Objective We hypothesized that aspirin desensitization blocks IL-4–induced expression of these LT activities through inhibition of signal transducer and activator of transcription 6 (STAT6)–mediated transcription. Methods Nuclear extracts were derived from THP-1 and normal human monocytes resting and cocultured with aspirin before IL-4 stimulation. Quantitative PCRs were conducted. Electrophoretic mobility shift assays were performed by using oligomers for STAT6 sites within the LT receptor and synthase promoters. Western blots of nuclear extracts were probed by using anti–phospho-STAT6 antibodies. Results Upregulation of LT receptor mRNA by IL-4 was negated by aspirin and ketorolac but not by sodium salicylate. The STAT6 site in the LT receptor and synthase promoters was confirmed by using mobility shift assays by competing with unlabeled STAT6 consensus probes and supershifts with anti-STAT6 antibodies. Aspirin and ketorolac decreased the IL-4–inducible expression of nuclear STAT6 observed in mobility shift assays and Western hybridization. Conclusion The LT receptor and synthase promoters have STAT6-binding sites that are engaged by IL-4–induced nuclear extracts and inhibited by aspirin. Assuming that normal monocytes behave like monocytes from patients with aspirin-exacerbated respiratory disease, inhibition of IL-4–STAT6 might explain a mechanism in aspirin desensitization daily treatment, resulting in downregulation of production and responsiveness to cysteinyl leukotrienes. This biologic activity of aspirin likely reflects COX inhibition because it was shared with ketorolac but not sodium salicylate.