Comparison of miRNA profiling during airway epithelial repair in undifferentiated and differentiated cells in vitro

Abstract

Respiratory epithelium is a highly integrated structure that efficiently protects lungs from extrinsic irritants thanks to rapid repair of the wound. The repair is a complex process that requires coordinated expression of networks of genes. Plausible regulators of this process are microRNAs. We investigated if global miRNA silencing influences the epithelial repair and if changes in miRNA expression profile during repair are similar between two bronchial epithelial cell cultures: differentiated and undifferentiated cells.Two bronchial cell types were used:16HBE14o-and NHBE. Transfection was performed with siRNAs against Drosha and Dicer. For miRNA profiling, non-transfected cells were cultured until confluent and harvested for RNA isolation at baseline (cells before wounding) and at different time post-wounding (8, 16, 24 and 48 hours). MicroRNA expression profiling was performed using TaqMan Array Human MicroRNA Card A. Target prediction was done in miRNA body map, and pathway analysis using DAVID.Cells with downregulated Drosha and Dicer demonstrated a significantly delayed wound repair in comparison to control in both cell lines. MiRNA expression profiling revealed that ten miRNAs exhibited significant changes over time after cell injury. These genes showed a similar expression pattern in both cell lines. The predicted targets of these miRNAs were then clustered by pathway analysis into six biological groups related to wound repair.Silencing of global miRNA expression confirmed that miRNAs are crucial for airway epithelial repair. Moreover, epithelial cells of two different origins demonstrated some similarities in miRNA expression pattern during wound repair independent of differentiation state.

Changes in miRNA Gene Expression during Wound Repair in Differentiated Normal Human Bronchial Epithelium

“…That corresponds to the results of our previous research [ 15 ], reporting changes in miRNA expression in undifferentiated cell line (16HBE14o-). Our recent findings [ 21 ] have also confirmed the importance of miRNA during this process: silencing of DICER and DROSHA , the enzymes crucial for RNA interference, resulted in significantly delayed wound repair. Profiling analysis of both cell types (undifferentiated and pseudostratified epithelium) showed some similarities between the expressions of several miRNA genes, suggesting their importance in the fundamental repair processes.…”

Targeting matrix metalloproteinase‐13 in bronchial epithelial repair

“…Additionally, the results were not confirmed in differentiated epithelial cells, using the air‐liquid interface (ALI) model. However, others have shown that NHBE in ALI close by only 20% even at 48 hours, which was suggested to be due to technical issues with using ALI for wound repair assays.…”

A database on differentially expressed microRNAs during rodent bladder healing

“…MicroRNAs have been intensively studied and today we understand that these molecules regulate several cellular and developmental pathways. MicroRNAs have shown differing expression patterns during tissue repair in bones 15 , 16 , lungs 17 skin 18 , 19 and oral mucosa 20 . These molecules mostly contribute to tissue repair by coordinating cell proliferation and inflammation among other functions 18 , 19 , 21 , 22 .…”