Transient Receptor Potential Ankyrin 1 Channels Modulate Inflammatory Response in Respiratory Cells from Patients with Cystic Fibrosis.
Pseudomonas aeruginosa colonization, prominent inflammation with massive expression of the neutrophil chemokine IL-8, and luminal infiltrates of neutrophils are hallmarks of chronic lung disease in patients with cystic fibrosis (CF). The nociceptive transient receptor potential ankyrin (TRPA) 1 calcium channels have been recently found to be involved in nonneurogenic inflammation. Here, we investigate the role of TRPA1 in CF respiratory inflammatory models in vitro. Expression of TRPA1 was evaluated in CF lung tissue sections and cells by immunohistochemistry and immunofluorescence. Epithelial cell lines (A549, IB3-1, CuFi-1, CFBE41o-) and primary cells from patients with CF were used to: (1) check TRPA1 function modulation, by Fura-2 calcium imaging; (2) down-modulate TRPA1 function and expression, by pharmacological inhibitors (HC-030031 and A-967079) and small interfering RNA silencing; and (3) assess the effect of TRPA1 down-modulation on expression and release of cytokines upon exposure to proinflammatory challenges, by quantitative RT-PCR and 27-protein Bioplex assay. TRPA1 channels are expressed in the CF pseudostratified columnar epithelium facing the bronchial lumina exposed to bacteria, where IL-8 is coexpressed. Inhibition of TRPA1 expression results in a relevant reduction of release of several cytokines, including IL-8 and the proinflammatory cytokines IL-1β and TNF-α, in CF primary bronchial epithelial cells exposed to P. aeruginosa and to the supernatant of mucopurulent material derived from the chronically infected airways of patients with CF. In conclusion, TRPA1 channels are involved in regulating the extent of airway inflammation driven by CF bronchial epithelial cells.
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Paola
P
Prandini
2 Department of Preclinical and Clinical Pharmacology, University of Florence, Florence, Italy.
Francesco
F
De Logu
2 Department of Preclinical and Clinical Pharmacology, University of Florence, Florence, Italy.
Camilla
C
Fusi
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Lisa
L
Provezza
2 Department of Preclinical and Clinical Pharmacology, University of Florence, Florence, Italy.
Romina
R
Nassini
3 Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
Giulia
G
Montagner
2 Department of Preclinical and Clinical Pharmacology, University of Florence, Florence, Italy.
Serena
S
Materazzi
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Silvia
S
Munari
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Eliana
E
Gilioli
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Valentino
V
Bezzerri
3 Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
Alessia
A
Finotti
3 Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
Ilaria
I
Lampronti
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Anna
A
Tamanini
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Maria Cristina
MC
Dechecchi
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Giuseppe
G
Lippi
4 Departments of Medicine and of Cell Biology and Physiology, Marsico Lung Institute, Cystic Fibrosis Research Center, University of North Carolina, Chapel Hill, North Carolina; and.
Carla M
CM
Ribeiro
5 Department of Morphology, Surgery, and Experimental Medicine, Section of Pathology, Oncology, and Experimental Biology, Laboratory for Technologies of Advanced Therapies, University of Ferrara, Ferrara, Italy.
Alessandro
A
Rimessi
5 Department of Morphology, Surgery, and Experimental Medicine, Section of Pathology, Oncology, and Experimental Biology, Laboratory for Technologies of Advanced Therapies, University of Ferrara, Ferrara, Italy.
Paolo
P
Pinton
3 Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
Roberto
R
Gambari
2 Department of Preclinical and Clinical Pharmacology, University of Florence, Florence, Italy.
Pierangelo
P
Geppetti
1 Laboratory of Molecular Pathology, Department of Pathology and Diagnostics, University Hospital, Verona, Italy.
Giulio
G
Cabrini
D000072283:A549 Cells; D000328:Adult; D001980:Bronchi / Q000473:pathology; D015220:Calcium Channels / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000150:complications* / Q000235:genetics / Q000473:pathology; D016207:Cytokines / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology; D005260:Female; D020868:Gene Silencing; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011014:Pneumonia / Q000150:complications* / Q000235:genetics / Q000473:pathology*; D011550:Pseudomonas aeruginosa / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000074025:TRPA1 Cation Channel; D014019:Tissue Donors; D014158:Transcription, Genetic; D050051:Transient Receptor Potential Channels / Q000037:antagonists & inhibitors / Q000378:metabolism*; D055815:Young Adult
D015220:Calcium Channels; D016207:Cytokines; D016209:Interleukin-8; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D000074025:TRPA1 Cation Channel; C473350:TRPA1 protein, human; D050051:Transient Receptor Potential Channels
IL-8; Pseudomonas aeruginosa; cystic fibrosis; inflammation; transient receptor potential ankyrin 1