Cold temperature induces mucin hypersecretion from normal human bronchial epithelial cells in vitro through a transient receptor potential melastatin 8 (TRPM8)–mediated mechanism

• Published in: Journal of allergy and clinical immunology Vol. 128; no. 3; pp. 626 - 634.e5
• Main Authors: Li, MinChao, Li, Qi, Yang, Gang, Kolosov, Victor P., Perelman, Juliy M., Zhou, Xiang Dong
• Format: Journal Article
• Language: English
• Published: New York, NY Mosby, Inc 01.09.2011; Elsevier
• Subjects: Aged; air; Allergy and Immunology; Animals; asthma; biochemical pathways; Biological and medical sciences; Bronchi - cytology; Bronchi - metabolism; Ca 2; calcium; Cell Line; cold; cold air; Cold Temperature; complementary DNA; enzyme-linked immunosorbent assay; epithelial cells; Epithelial Cells - metabolism; epithelium; Female; fluorescent proteins; Fundamental and applied biological sciences. Psychology; Fundamental immunology; gene expression; Gene Expression Regulation; Humans; hydrolysis; hypersecretion; image analysis; immunohistochemistry; Immunopathology; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Male; Medical sciences; Membrane Proteins - genetics; Membrane Proteins - metabolism; messenger RNA; Middle Aged; MUC5AC; Mucin 5AC - genetics; Mucin 5AC - metabolism; mucins; Mucins - metabolism; mucus; mucus hypersecretion; Myristoylated Alanine-Rich C Kinase Substrate; patients; phosphatidylinositol 4,5-bisphosphate; phospholipase C; phosphorylation; Pulmonary Disease, Chronic Obstructive - metabolism; quantitative polymerase chain reaction; reverse transcriptase polymerase chain reaction; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; serine; signal transduction; temperature; Transient receptor potential melastatin 8; TRPM Cation Channels - genetics; TRPM Cation Channels - metabolism; Western blotting; PLC; cold air; Transient receptor potential melastatin 8; Ca 2; GFP; TRP; shRNA; NHBE; TRPM8; PSD; PKC; ER; IP3; MUC5AC; BCTC; mucus hypersecretion; FVC; MARCKS; myristoylated alanine-rich C kinase substrate; phosphatidylinositol 4,5-bisphosphate; PH; GAPDH; PIP2; COPD; ERK; Protein kinase C; Forced vital capacity; Normal human bronchial epithelial; Pleckstrin homology; Chronic obstructive pulmonary disease; Inositol trisphosphate; N-(4-tert-butylphenyl)-4-(3-chloropyridin-2-yl) piperazine-1-carboxamide; Extracellular signal-regulated kinase; Transient receptor potential; Phospholipase C; Green fluorescent protein; Glyceraldehyde-3-phosphate dehydrogenase; Phosphorylation site domain; Short hairpin RNA; Endoplasmic reticulum; Temperature; Calcium; Environmental factor; Mucus; Respiratory system; Inorganic element; Mechanism; Respiratory tract; Phosphatidylinositol; Immunology; Biological receptor; Human; Immunopathology; Alanine; Cold; Enzyme; Mucin 5AC; Transferases; Bronchus; +; Hypersecretion; In vitro; Transitory; Substrate; Cold air; Kinase; Aminoacid; Epithelial cell; Ca
• ISSN: 0091-6749; 1097-6825; 1097-6825
• DOI: 10.1016/j.jaci.2011.04.032

Cold air stimulus is a major environmental factor that exacerbates chronic inflammatory airway diseases, such as chronic obstructive pulmonary disease (COPD) and asthma. At the molecular level, cold is detected by transient receptor potential melastatin 8 (TRPM8). To date, TRPM8 expression has not been characterized in the airway epithelium of patients with COPD. The role of TRPM8 channels in a series of airway responses induced by cold stimuli and the molecular and biochemical pathways of TRPM8 in regulating cold-induced responses are largely unknown. We sought to explore the role of TRPM8 in cold air–provoked mucus hypersecretion and the potential signaling pathway involved in this process. The expression of TRPM8 in the bronchial epithelium was examined by means of immunohistochemistry, RT-PCR, and Western blotting. TRPM8 receptor function and hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) were characterized by means of Ca 2+ imaging and spatiotemporal dynamics of phospholipase C (PLC) δ1–pleckstrin homology–green fluorescent protein, respectively. The expression of MUC5AC mRNA and MUC5AC mucin protein was measured by using real-time PCR and ELISA, respectively. Four serine residues in the myristoylated alanine-rich C kinase substrate (MARCKS)–phosphorylation site domain were mutated to identify the function of MARCKS in TRPM8-mediated airway mucus hypersecretion. TRPM8 protein and mRNA expression were significantly increased in patients with COPD compared with expression seen in healthy subjects. Cold produced robust increases in intracellular Ca 2+ levels and promoted translocation of PLCδ1–pleckstrin homology–green fluorescent protein. Cold increased expression of MUC5AC mRNA and intracellular and secreted MUC5AC protein in a nonsustained way. Phosphorylation site domain–mutant MARCKS cDNA hindered MUC5AC secretion induced by cold. These results indicate that the TRPM8 receptor is involved in cold-induced mucus hypersecretion through the Ca 2+-PLC-PIP2-MARCKS signaling pathway.