snoRNAs Downregulate Smooth Muscle Cell COX4I2 and Promote Neointimal Hyperplasia.

BACKGROUND

Reactive oxygen species (ROS) augment the activation of vascular smooth muscle cells (SMCs) and promote neointimal hyperplasia evoked by arterial injury or atherogenesis. We have previously shown that small nucleolar RNAs (snoRNAs) from the locus are key regulators of cellular ROS levels.

METHODS

Using mice deficient in the snoRNAs, we tested whether these snoRNAs regulate SMC activation in vitro and in vivo. Carotid endothelial denudation was used to provoke neointimal hyperplasia in wild-type (WT) and snoRNA knockout (snoKO) mice, which lack all four intronically-encoded snoRNAs. Primary SMCs from WT and snoKO mice were used for in vitro functional and proteomic analyses. HEK293T cells with specific snoRNA deletions were used to test for snoRNA-guided 2'- -methylation of mRNA.

RESULTS

Arterial ROS levels, inflammation, and carotid artery neointimal hyperplasia were reduced in snoKO compared with WT mice. In vitro, snoKO SMCs demonstrated lower ROS levels and less migration, proliferation, and inflammatory signaling than WT SMCs. Reduced ROS levels in snoKO SMCs and aortas correlated with upregulation of the mitochondrial protein COX4I2, which is associated with reduced mitochondrial ROS under normoxic conditions. Deleting the snoRNA in human HEK293T cells decreased 2'- -methylation of mRNA and upregulated COX4I2 protein without changing COX4I2 mRNA levels. Silencing in snoKO SMCs upregulated SMC ROS to WT levels.

CONCLUSIONS

snoRNAs are important drivers of SMC activation and neointimal hyperplasia. snoRNAs augment SMC ROS levels, at least in part, by post-transcriptional downregulation of COX4I2 expression.