The Mechanosensitive microRNA‐712, Induced by Disturbed Blood Flow in Endothelium, Plays a Key Role in Atherosclerosis

Abstract

Atherosclerosis is the underlying cause of heart attack and stroke and preferentially occurs in arterial regions exposed to disturbed flow (d-flow) by mechanisms involving broad changes in gene expression. While miRNAs are known to regulate various aspects of cardiovascular biology and disease, their role in atherosclerosis is unclear. Here, we identified novel mechanosensitive miRNAs using the same mouse model and endothelial miRNA array. Of those, we identified miR-712 as the most shear-sensitive miRNA induced by d-flow both in vivo and in vitro. By integrated systems biological approach, we developed a gene interactome map, predicting that miR-712 regulates a key gene hub-TIMP3, known to govern matrix metalloproteinases and atherosclerosis. Using gain-of-function (pre-miR-712) and loss-of-function [locked nucleic acid (LNA)-based anti-miR-712] studies, we experimentally validated and showed that miR-712 directly regulates expression of TIMP3 in a flow-dependent manner Further, we showed that inhibition of mechanosensitive miRNA by subcutaneous injection of anti-miR-712 prevented atherosclerosis in ApoE-/- mice These findings highlight how an integrative approach, combining systems biology with functional genomics in a single experimental animal model can be used to overcome a major obstacle in miRNA research: linking individual miRNA changes to a network of potential target genes in an unbiased manner to identify relevant molecular pathways and generate specific, testable hypotheses. Further, our results strongly suggest that targeting mechanosensitive “athero-miR” with anti-miR-based approaches may provide a new treatment paradigm in atherosclerosis.