Molecular control of nitric oxide synthesis through eNOS and caveolin-1 interaction regulates osteogenic differentiation of adipose-derived stem cells by modulation of Wnt/?-catenin signaling.

ABSTRACT: Nitric oxide (NO) plays a role in a number of physiological processes including stem cell differentiation and osteogenesis. Endothelial nitric oxide synthase (eNOS), one of three NO-producing enzymes, is located in a close conformation with the caveolin-1 (CAV-1WT) membrane protein which is inhibitory to NO production. Modification of this interaction through mutation of the caveolin scaffold domain can increase NO release. In this study, we genetically modified equine adipose-derived stem cells (eASCs) with eNOS, CAV-1WT, and a CAV-1F92A (CAV-1WT mutant) and assessed NO-mediated osteogenic differentiation and the relationship with the Wnt signaling pathway.NO production was enhanced by lentiviral vector co-delivery of eNOS and CAV-1F92A to eASCs, and osteogenesis and Wnt signaling was assessed by gene expression analysis and activity of a novel Runx2-GFP reporter. Cells were also exposed to a NO donor (NONOate) and the eNOS inhibitor, L-NAME.NO production as measured by nitrite was significantly increased in eNOS and CAV-1F92A transduced eASCs +(5.59?±?0.22 ?M) compared to eNOS alone (4.81?±?0.59 ?M) and un-transduced control cells (0.91?±?0.23 ?M) (p?

SUBMITTER: Bandara N 

PROVIDER: S-EPMC5142348 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature