Abstract
Cystic fibrosis results from reduced cystic fibrosis transmembrane conductance regulator protein activity leading to defective epithelial ion transport. Ca2+-activated Cl− channels mediate physiological functions independently of cystic fibrosis transmembrane conductance regulator. Anoctamin 1 (ANO1/TMEM16A) was identified as the major Ca2+-activated Cl− channel in airway epithelial cells, and we recently demonstrated that downregulation of the anoctamin 1 channel in cystic fibrosis patients contributes to disease severity via an unknown mechanism. Here we show that microRNA-9 (miR-9) contributes to cystic fibrosis and downregulates anoctamin 1 by directly targeting its 3′UTR. We present a potential therapy based on blockage of miR-9 binding to the 3′UTR by using a microRNA target site blocker to increase anoctamin 1 activity and thus compensate for the cystic fibrosis transmembrane conductance regulator deficiency. The target site blocker is tested in in vitro and in mouse models of cystic fibrosis, and could be considered as an alternative strategy to treat cystic fibrosis.
Highlights
Cystic fibrosis results from reduced cystic fibrosis transmembrane conductance regulator protein activity leading to defective epithelial ion transport
We propose an alternative strategy to correct chloride efflux in Cystic fibrosis (CF) patients that rely on a target site blocker (TSB) that prevents miR-9 targeting of anoctamin 1 (ANO1)
After preliminary luciferase assays to test the binding of these miRNAs to the ANO1 3′UTR, we focused on miR-9. miR-9 expression was studied in CF (CFBE41o-) and non-CF (16HBE14o-) bronchial epithelial cells by qRT-PCR and showed a 2.5-fold increase in CF cells as compared to non-CF cells (Fig. 1a)
Summary
Cystic fibrosis results from reduced cystic fibrosis transmembrane conductance regulator protein activity leading to defective epithelial ion transport. We present a potential therapy based on blockage of miR-9 binding to the 3′UTR by using a microRNA target site blocker to increase anoctamin 1 activity and compensate for the cystic fibrosis transmembrane conductance regulator deficiency. We show that miR-9 is overexpressed in CF cells where it directly regulates ANO1, causing a decrease in its expression and activity Based on this knowledge, we propose an alternative strategy to correct chloride efflux in CF patients that rely on a target site blocker (TSB) that prevents miR-9 targeting of ANO1. We propose an alternative strategy to correct chloride efflux in CF patients that rely on a target site blocker (TSB) that prevents miR-9 targeting of ANO1 This therapeutic approach results in increased chloride efflux mediated by ANO1, mucociliary clearance, and migration rate of cells in in vitro and in vivo CF models
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