Calcific aortic valve disease (CAVD) is a major cause of morbidity in the aging population, but the underlying mechanisms of its progression remain poorly understood. Aortic valve calcification preferentially occurs on the fibrosa, which is subjected to disturbed flow. The side-specific progression of the disease is characterized by inflammation, calcific lesions, and extracellular matrix (ECM) degradation. Here, we explored the role of mechanosensitive microRNA-181b and its downstream targets in human aortic valve endothelial cells (HAVECs). Mechanistically, miR-181b is upregulated in OS and fibrosa, and it targets TIMP3, SIRT1, and GATA6, correlated with increased gelatinase/MMP activity. Overexpression of miR-181b led to decreased TIMP3 and exacerbated MMP activity as shown by gelatinase assay, and miR-181b inhibition decreased gelatinase activity through the repression of TIMP3 levels. Luciferase assay showed specific binding of miR-181b to the TIMP3 gene. Overexpression of miR-181b in HAVECs subjected to either LS or OS increased MMP activity, and miR-181b inhibition abrogated shear-sensitive MMP activity. These studies suggest that targeting this shear-dependent miRNA may provide a novel noninvasive treatment for CAVD.
This is a preview of subscription content, log in to check access.
This study was funded by NIH (R01HL114772) as well as NHLBI Grants (HL119798, HL113451, HL095070 and HL124879).
Conflict of Interest
All the authors declare that they have no conflict of interest.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
Supplementary Table IList of quantitative PCR primers used in analysis of gene targets of miRNA-181b (DOCX 112 kb)
Alfonso-Jaume, M. A., et al. Cardiac ischemia-reperfusion injury induces matrix metalloproteinase-2 expression through the AP-1 components FosB and JunB. Am. J. Physiol. Heart Circ. Physiol. 291:H1838–1846, 2006. doi:10.1152/ajpheart.00026.2006.CrossRefGoogle Scholar
Balachandran, K., P. Sucosky, H. Jo, and A. P. Yoganathan. Elevated cyclic stretch alters matrix remodeling in aortic valve cusps: implications for degenerative aortic valve disease. Am. J. Physiol. Heart Circ. Physiol. 296:H756–764, 2009. doi:10.1152/ajpheart.00900.2008.CrossRefGoogle Scholar
Chen, J. H., and C. A. Simmons. Cell-matrix interactions in the pathobiology of calcific aortic valve disease: critical roles for matricellular, matricrine, and matrix mechanics cues. Circ. Res. 108:1510–1524, 2011. doi:10.1161/CIRCRESAHA.110.234237.CrossRefGoogle Scholar
Chen, Y. X., M. Zhang, Y. Cai, Q. Zhao, and W. Dai. The Sirt1 activator SRT1720 attenuates angiotensin II-induced atherosclerosis in apoE(−)/(−) mice through inhibiting vascular inflammatory response. Biochem. Biophys. Res. Commun. 465:732–738, 2015. doi:10.1016/j.bbrc.2015.08.066.CrossRefGoogle Scholar
Cheung, P. Y., et al. Matrix metalloproteinase-2 contributes to ischemia-reperfusion injury in the heart. Circulation 101:1833–1839, 2000.CrossRefGoogle Scholar
Holliday, C. J., R. F. Ankeny, H. Jo, and R. M. Nerem. Discovery of shear- and side-specific mRNAs and miRNAs in human aortic valvular endothelial cells. Am. J. Physiol. Heart Circ. Physiol. 301:H856–867, 2011. doi:10.1152/ajpheart.00117.2011.CrossRefGoogle Scholar
Hsu, S. Y., I. C. Hsieh, S. H. Chang, M. S. Wen, and K. C. Hung. Aortic valve sclerosis is an echocardiographic indicator of significant coronary disease in patients undergoing diagnostic coronary angiography. Int. J. Clin. Pract. 59:72–77, 2005. doi:10.1111/j.1742-1241.2004.00219.x.CrossRefGoogle Scholar
Kim, C. W., et al. Prevention of abdominal aortic aneurysm by anti-microRNA-712 or anti-microRNA-205 in angiotensin II-infused mice. Arterioscler. Thromb. Vasc. Biol. 34:1412–1421, 2014. doi:10.1161/ATVBAHA.113.303134.CrossRefGoogle Scholar
Kumar, S., C. W. Kim, R. D. Simmons, and H. Jo. Role of flow-sensitive microRNAs in endothelial dysfunction and atherosclerosis: mechanosensitive athero-miRs. Arterioscler. Thromb. Vasc. Biol. 34:2206–2216, 2014. doi:10.1161/atvbaha.114.303425.CrossRefGoogle Scholar
Lin, J., et al. MicroRNA-181b inhibits thrombin-mediated endothelial activation and arterial thrombosis by targeting caspase recruitment domain family member 10. FASEB J. 2016. doi:10.1096/fj.201500163R.Google Scholar
Mohler, E. R., M. J. Sheridan, R. Nichols, W. P. Harvey, and B. F. Waller. Development and progression of aortic-valve stenosis—atherosclerosis risk-factors—a causal relationship—a clinical morphological-study. Clin. Cardiol. 14:995–999, 1991.CrossRefGoogle Scholar
Mohler, E. R., et al. Bone formation and inflammation in cardiac valves. Circulation 103:1522–1528, 2001.CrossRefGoogle Scholar
Ni, C. W., H. Qiu, and H. Jo. MicroRNA-663 upregulated by oscillatory shear stress plays a role in inflammatory response of endothelial cells. Am. J. Physiol. Heart Circ. Physiol. 300:H1762–1769, 2011. doi:10.1152/ajpheart.00829.2010.CrossRefGoogle Scholar
Otto, C. M., J. Kuusisto, D. D. Reichenbach, A. M. Gown, and K. D. Obrien. Characterization of the early lesion of degenerative valvular aortic-stenosis—histological and immunohistochemical studies. Circulation 90:844–853, 1994.CrossRefGoogle Scholar
Perrotta, I., A. Sciangula, S. Aquila, and S. Mazzulla. Matrix metalloproteinase-9 expression in calcified human aortic valves: a histopathologic, immunohistochemical, and ultrastructural study. Appl. Immunohistochem. Mol. Morphol. 24:128–137, 2016. doi:10.1097/pai.0000000000000144.CrossRefGoogle Scholar
Platt, M. O., Y. Xing, H. Jo, and A. P. Yoganathan. Cyclic pressure and shear stress regulate matrix metalloproteinases and cathepsin activity in porcine aortic valves. J. Heart Valve Dis. 15:622–629, 2006.Google Scholar
Schonbeck, U., et al. Expression of stromelysin-3 in atherosclerotic lesions: regulation via CD40-CD40 ligand signaling in vitro and in vivo. J. Exp. Med. 189:843–853, 1999.CrossRefGoogle Scholar
Son, D. J., et al. The atypical mechanosensitive microRNA-712 derived from pre-ribosomal RNA induces endothelial inflammation and atherosclerosis. Nat. Commun. 4:3000, 2013. doi:10.1038/ncomms4000.CrossRefGoogle Scholar
Soumyarani, V. S., and N. Jayakumari. Oxidatively modified high density lipoprotein promotes inflammatory response in human monocytes-macrophages by enhanced production of ROS, TNF-alpha, MMP-9, and MMP-2. Mol. Cell. Biochem. 366:277–285, 2012. doi:10.1007/s11010-012-1306-y.CrossRefGoogle Scholar
Stephens, E. H., and K. J. Grande-Allen. Age-related changes in collagen synthesis and turnover in porcine heart valves. J. Heart Valve Dis. 16:672–682, 2007.Google Scholar
Sucosky, P., K. Balachandran, A. Elhammali, H. Jo, and A. P. Yoganathan. Altered shear stress stimulates upregulation of endothelial VCAM-1 and ICAM-1 in a BMP-4- and TGF-β1-dependent pathway. Arterioscler. Thromb. Vasc. Biol. 29:254–260, 2009. doi:10.1161/atvbaha.108.176347.CrossRefGoogle Scholar
Sun, X., et al. MicroRNA-181b regulates NF-kappaB-mediated vascular inflammation. J. Clin. Investig. 122:1973–1990, 2012. doi:10.1172/JCI61495.Google Scholar
Uzui, H., et al. Increased expression of membrane type 3-matrix metalloproteinase in human atherosclerotic plaque: role of activated macrophages and inflammatory cytokines. Circulation 106:3024–3030, 2002.CrossRefGoogle Scholar
Wang, Y., et al. Circulating matrix metalloproteinase patterns in association with aortic dilatation in bicuspid aortic valve patients with isolated severe aortic stenosis. Heart Vessels 31:189–197, 2016. doi:10.1007/s00380-014-0593-5.CrossRefGoogle Scholar
Yip, C. Y., J. H. Chen, R. Zhao, and C. A. Simmons. Calcification by valve interstitial cells is regulated by the stiffness of the extracellular matrix. Arterioscler. Thromb. Vasc. Biol. 29:936–942, 2009. doi:10.1161/ATVBAHA.108.182394.CrossRefGoogle Scholar