SAT-LB97 MiRNA-99a and mTOR2 Mediate Enhanced Endothelial Mineralocorticoid Receptor Signaling-Induced Activation of Sodium Channel and Endothelium Stiffness

In diet induced obesity enhanced endothelial cell (EC) mineralocorticoid receptor (MR) (ECMR) and downstream sodium channel (EnNaC) activity increases oxidative stress and inflammation, thereby promoting vascular stiffness and associated impaired endothelial mediated relaxation. For example, consumption of a Western diet (WD) containing excess fat (46%) and fructose (17.5%) for 16 weeks elevated plasma aldosterone levels and increased vascular MR expression in conjunction with increased endothelial and vascular stiffness in female mice. EC specific deletion of either the ECMR or EnNaC significantly attenuated this diet induced endothelial/vascular stiffness. Emerging information suggests that abnormal expression of miR-99a may be involved in these processes. To this point, we recently observed that aldosterone (10(-7) mol/L) causes a reduction in miR-99a that was prevented by the MR antagonist, spironolactone (10µM) in in vitro ECs. By using RNA sequencing, we also demonstrated that ECMR activation reduced arterial miR-99a expression in diet induced obesity. Since the mammalian target of rapamycin (mTOR2)/SGK1 signaling pathway is involved in aldosterone activation of ENaC we then explored the effects of miR-99a on mTOR2 expression. Indeed, miR-99a reduced mTOR2. We further observed that inhibition of mTOR2 with PP242 inhibited EnNaC activity as determined by patch clamping of ECs. Collectively these data suggest that consumption of a WD induced ECMR activation and increased EnNaC activity and endothelial stiffness, in part, by reducing the tonic inhibitory effects exerted by miR-99a on mTOR2 mediated EnNaC activation.