title: Oxidative-mechanical stress signals stem cell niche mediated Lrp5 osteogenesis in eNOS(-/-)  null mice.
abstract: Calcific aortic valve disease (CAVD) is the most common indication for valve surgery in the USA. This study hypothesizes that CAVD develops secondary to Wnt3a/Lrp5 activation via oxidative-mechanical stress in eNOS null mice. eNOS(-/-) mice were tested with experimental diets including a control (n=20), cholesterol (n=20), cholesterol + Atorvastatin (n=20). After 23 weeks the mice were tested for the development of aortic stenosis by Echo, Histology, MicroCT, and RTPCR for bone markers. In vitro studies measured Wnt3a secretion from aortic valve endothelial cells and confirmed oxidative stress via eNOS activity. Anion exchange chromatography was performed to isolate the mitogenic protein. Myofibroblast cells were tested to induce bone formation. Cholesterol treated eNOS mice develop severe stenosis with an increase in Wnt3a, Lrp5, Runx2 (threefold increase (P<0.0001) in the bicuspid versus tricuspid aortic valves. Secretion of Wnt3a from aortic valve endothelium in the presence of abnormal oxidative stress was correlated with diminished eNOS enzymatic activity and tissue nitrite levels. Initial characterization of the architecture for a stem cell nice was determined by protein isolation using anion-exchange chromatography and cell proliferation via thymidine incorporation. Osteoblastogenesis in the myofibroblast cell occurred via Lrp5 receptor upregulation in the presence of osteogenic media. Targeting the Wnt3a/Lrp5 pathway in valve calcification and activation of osteogenesis is via an oxidative-mechanical stress in CAVD. These findings provide a foundation for treating this disease process by targeting the cross talk mechanism in a resident stem cell niche.
authors.affiliation: Division of Cardiology, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA. nrajamannan@gmail.com
authors.forename: Nalini M
authors.initials: NM
authors.lastname: Rajamannan
mesh_terms: D000818:Animals; D001021:Aortic Valve / Q000002:abnormalities / Q000378:metabolism / Q000473:pathology; D001024:Aortic Valve Stenosis / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D002114:Calcinosis / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D002450:Cell Communication; D002454:Cell Differentiation; D002791:Cholesterol, Dietary / Q000008:administration & dosage; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology; D006801:Humans; D060492:Low Density Lipoprotein Receptor-Related Protein-5 / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008955:Models, Cardiovascular; D058628:Myofibroblasts / Q000378:metabolism / Q000473:pathology; D052250:Nitric Oxide Synthase Type III / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology*; D018384:Oxidative Stress; D015398:Signal Transduction; D055153:Stem Cell Niche / Q000502:physiology*; D013314:Stress, Mechanical; D060509:Wnt3A Protein / Q000378:metabolism
chemical_list: D002791:Cholesterol, Dietary; D050659:Core Binding Factor Alpha 1 Subunit; D060492:Low Density Lipoprotein Receptor-Related Protein-5; C555483:Lrp5 protein, mouse; C493716:Runx2 protein, mouse; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse