TY - JOUR T1 - Phospholipase C-β3 is a key modulator of IL-8 expression in cystic fibrosis bronchial epithelial cells. JF - J Immunol Y1 - 2011 A1 - Bezzerri, Valentino A1 - d'Adamo, Pio A1 - Rimessi, Alessandro A1 - Lanzara, Carmen A1 - Crovella, Sergio A1 - Nicolis, Elena A1 - Tamanini, Anna A1 - Athanasakis, Emmanouil A1 - Tebon, Maela A1 - Bisoffi, Giulia A1 - Drumm, Mitchell L A1 - Knowles, Michael R A1 - Pinton, Paolo A1 - Gasparini, Paolo A1 - Berton, Giorgio A1 - Cabrini, Giulio KW - Adenosine Triphosphate KW - Calcium KW - Cell Line, Transformed KW - Cystic Fibrosis KW - Enzyme Activation KW - Epithelial Cells KW - Gene Expression KW - Gene Frequency KW - Genotype KW - Green Fluorescent Proteins KW - Host-Pathogen Interactions KW - Humans KW - Interleukin-8 KW - Isoenzymes KW - Lung Diseases KW - Microscopy, Fluorescence KW - Phospholipase C beta KW - Polymorphism, Single Nucleotide KW - Protein Kinase C KW - Protein Kinase C beta KW - Pseudomonas aeruginosa KW - RNA Interference KW - Toll-Like Receptors KW - Transcription Factor RelA AB -

Respiratory insufficiency is the major cause of morbidity and mortality in patients affected by cystic fibrosis (CF). An excessive neutrophilic inflammation, mainly orchestrated by the release of IL-8 from bronchial epithelial cells and amplified by chronic bacterial infection with Pseudomonas aeruginosa, leads to progressive tissue destruction. The anti-inflammatory drugs presently used in CF patients have several limitations, indicating the need for identifying novel molecular targets. To address this issue, we preliminarily studied the association of 721 single nucleotide polymorphisms from 135 genes potentially involved in signal transduction implicated in neutrophil recruitment in a cohort of F508del homozygous CF patients with either severe or mild progression of lung disease. The top ranking association was found for a nonsynonymous polymorphism of the phospholipase C-β3 (PLCB3) gene. Studies in bronchial epithelial cells exposed to P. aeruginosa revealed that PLCB3 is implicated in extracellular nucleotide-dependent intracellular calcium signaling, leading to activation of the protein kinase Cα and Cβ and of the nuclear transcription factor NF-κB p65. The proinflammatory pathway regulated by PLCB3 acts by potentiating the Toll-like Receptors' signaling cascade and represents an interesting molecular target to attenuate the excessive recruitment of neutrophils without completely abolishing the inflammatory response.

VL - 186 IS - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21411730?dopt=Abstract ER - TY - JOUR T1 - A polymorphism in the 5' UTR of the DEFB1 gene is associated with the lung phenotype in F508del homozygous Italian cystic fibrosis patients. JF - Clin Chem Lab Med Y1 - 2011 A1 - Crovella, Sergio A1 - Segat, Ludovica A1 - Amato, Annalisa A1 - Athanasakis, Emmanouil A1 - Bezzerri, Valentino A1 - Braggion, Cesare A1 - Casciaro, Rosaria A1 - Castaldo, Giuseppe A1 - Colombo, Carla A1 - Covone, Angela Elvira A1 - De Rose, Virginia A1 - Gagliardini, Rolando A1 - Lanzara, Carmen A1 - Minicucci, Laura A1 - Morgutti, Marcello A1 - Nicolis, Elena A1 - Pardo, Francesca A1 - Quattrucci, Serena A1 - Raia, Valeria A1 - Ravazzolo, Roberto A1 - Seia, Manuela A1 - Stanzial, Valentino A1 - Termini, Lisa A1 - Zazzeron, Laura A1 - Cabrini, Giulio A1 - Gasparini, Paolo KW - 5' Untranslated Regions KW - Adult KW - beta-Defensins KW - Cystic Fibrosis KW - Cystic Fibrosis Transmembrane Conductance Regulator KW - Female KW - Genotype KW - Homozygote KW - Humans KW - Italy KW - Male KW - Mutation KW - Phenotype KW - Polymorphism, Genetic KW - Young Adult AB -

BACKGROUND: The identification of cystic fibrosis (CF) patients who are at greater risk of lung damage could be clinically valuable. Thus, we attempted to replicate previous findings and verify the possible association between three single nucleotide polymorphisms (SNPs c.-52G>A, c.-44C>G and c.-20G>A) in the 5' untranslated region (5' UTR) of the β defensin 1 (DEFB1) gene and the CF pulmonary phenotype.

METHODS: Genomic DNA from 92 Italian CF patients enrolled in different regional CF centres was extracted from peripheral blood and genotyped for DEFB1 SNPs using TaqMan(®) allele specific probes. In order to avoid genetic confounding causes that can account for CF phenotype variability, all patients were homozygous for the F508del CFTR mutation, and were then classified on the basis of clinical and functional data as mild lung phenotype (Mp, n=50) or severe lung phenotype patients (Sp, n=42).

RESULTS: For the c.-20G>A SNP, the frequency of the A allele, as well as the AA genotype, were significantly more frequent in Mp than in Sp patients, and thus this was associated with a protective effect against severe pulmonary disease (OR=0.48 and 0.28, respectively). The effect of the c.-20G>A A allele is consistent with a recessive model, and the protective effect against Sp is exerted only when it is present in homozygosis. For the other two SNPs, no differences were observed as allelic and genotypic frequency in the two subgroups of CF patients.

CONCLUSIONS: Our results, although necessary to be confirmed in larger and multiethnic populations, reinforce DEFB1 as a candidate modifier gene of the CF pulmonary phenotype.

VL - 49 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21077791?dopt=Abstract ER -