Type

Journal Article

Authors

Catherine M Greene
Noel G McElvaney
Sanjay H Chotirmall
Irene K Oglesby

Subjects

Biochemistry

Topics
pseudomonas aeruginosa base sequence microbiology gene expression profiling 3 untranslated regions cell line adult lung mirn145 microrna human humans cystic fibrosis transmembrane conductance regulator metabolism gene expression respiratory mucosa cystic fibrosis gene expression regulation female chemistry micrornas genotype genetics pathology chlorides gene knockdown techniques male intracellular space mirn223 microrna human mirn494 microrna human

Regulation of cystic fibrosis transmembrane conductance regulator by microRNA-145, -223, and -494 is altered in ΔF508 cystic fibrosis airway epithelium. (2013)

Abstract Expression of the cystic fibrosis transmembrane conductance regulator (CFTR) is altered in individuals with the ΔF508 CFTR mutation. We previously reported differential expression of microRNA (miRNA) in CF airway epithelium; however, the role of miRNA in regulation of CFTR expression here remains unexplored. In this study, we investigated the role of upregulated miRNAs in CFTR regulation in vivo in bronchial brushings from individuals homozygous or heterozygous for ΔF508 CFTR, validated our observations in vitro, and assessed the impact of defective chloride ion conductance, genotype, and colonization status on miRNA expression. miRNA target prediction was performed in silico, and expression of miRNA and target genes were measured by quantitative real-time PCR and/or Western blotting. Overexpression and inhibition studies were performed with pre-miRs or antimiRs, respectively, and a luciferase reporter gene was used to elucidate direct miRNA-target interactions. miR-145, miR-223, and miR-494 were upregulated in CF versus non-CF bronchial brushings and cell lines; in ΔF508 CFTR homozygotes versus heterozygotes; in subjects positive for P. aeruginosa; and in cells treated with a CFTR inhibitor or IL-1β. Reciprocal downregulation or upregulation of CFTR gene and/or protein expression was observed after miRNA manipulation and direct miRNA-target relationships demonstrated via a reporter system containing a wild type or mutated full-length CFTR 3' untranslated region. Increased expression of miR-145, miR-223, and miR-494 in vivo in bronchial epithelium of individuals carrying the ΔF508 CFTR mutation correlates with decreased CFTR expression. Defective CFTR function, Pseudomonas colonization, and inflammation may affect miRNA expression and contribute to the regulation of ΔF508 CFTR.
Collections Ireland -> Royal College of Surgeons in Ireland -> PubMed

Full list of authors on original publication

Catherine M Greene, Noel G McElvaney, Sanjay H Chotirmall, Irene K Oglesby

Experts in our system

1
Catherine M Greene
Royal College of Surgeons in Ireland
Total Publications: 150
 
2
Noel G McElvaney
Royal College of Surgeons in Ireland
Total Publications: 194
 
3
Sanjay H Chotirmall
Royal College of Surgeons in Ireland
Total Publications: 22