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MicroRNA profiling of cystic fibrosis intestinal disease in mice
Mark Bazett1, Alexandra Paun, Christina K Haston
1Meakins-Christie Laboratories and the Department of Human Genetics, McGill University, Montreal, PQ, Canada.
Abstract:
Cystic fibrosis (CF) intestinal disease is characterized by alterations in processes such as proliferation and apoptosis which are known to be regulated in part by microRNAs. Herein, we completed microRNA expression profiling of the intestinal tissue from the cystic fibrosis mouse model of cystic fibrosis transmembrane conductance regulator (Cftr) deficient mice (BALBc/J Cftr(tm1UNC)), relative to that of wildtype littermates, to determine whether changes in microRNA expression level are part of this phenotype. We identified 24 microRNAs to be significantly differentially expressed in tissue from CF mice compared to wildtype, with the higher expression in tissue from CF mice. These data were confirmed with real time PCR measurements. A comparison of the list of genes previously reported to have decreased expression in the BALB×C57BL/6J F2 CF intestine to that of genes putatively targeted by the 24 microRNAs, determined from target prediction software, revealed 155 of the 759 genes of the expression profile (20.4%) to overlap with predicted targets, which is significantly more than the 100 genes expected by chance (p=1×10(-8)). Pathway analysis identified these common genes to function in phosphatase and tensin homolog-, protein kinase A-, phosphoinositide-3 kinase/Akt- and peroxisome proliferator-activated receptor alpha/retinoid X receptor alpha signaling pathways, among others, and through real time PCR experiments genes of these pathways were demonstrated to have lower expression in the BALB CF intestine. We conclude that altered microRNA expression is a feature which putatively influences both metabolic abnormalities and the altered tissue homeostasis component of CF intestinal disease.
Insights
Altered microRNA expression in cystic fibrosis (CF) intestinal tissue impacts key signaling pathways. These microRNA changes may influence metabolic dysfunction and tissue imbalance in CF intestinal disease.
Area of Science:
- Molecular Biology
- Genetics
- Gastroenterology
Background:
- Cystic fibrosis (CF) intestinal disease involves altered cellular processes like proliferation and apoptosis.
- MicroRNAs are known regulators of these cellular processes.
Purpose of the Study:
- To investigate microRNA expression profiles in the intestinal tissue of a cystic fibrosis mouse model.
- To determine if altered microRNA levels contribute to the intestinal phenotype in CF.
Main Methods:
- MicroRNA expression profiling of intestinal tissue from CF transmembrane conductance regulator (Cftr) deficient mice and wildtype littermates.
- Validation of differentially expressed microRNAs using real-time PCR.
- Bioinformatic analysis to identify predicted microRNA targets and associated signaling pathways.
Main Results:
- Identified 24 significantly differentially expressed microRNAs in CF mouse intestinal tissue.
- Found a significant overlap between predicted microRNA targets and genes with decreased expression in CF intestine.
- Pathway analysis revealed involvement of PTEN, PKA, PI3K/Akt, and PPARα/RXRα signaling pathways.
Conclusions:
- Altered microRNA expression is a characteristic feature of CF intestinal disease.
- These microRNA changes potentially influence metabolic abnormalities and tissue homeostasis in the CF intestine.
