Abstract 296: The RNA-Binding Protein Quaking Maintains Endothelial Barrier Function Through Targeting Vascular Endothelial Cadherin mRNA
Endothelial cells (ECs) form a vital barrier between the blood and the artery wall, and play a major role in the onset of atherosclerosis. Endothelial monolayer integrity is determined largely by EC-cell interactions, where vascular endothelial cadherin (VE-cadherin) is the central adhesive component at these endothelial adherens junctions. While many of the receptors and signaling proteins involved in regulating these adherens junctions have been identified, surprisingly little is known regarding their regulation at the post[[Unable to Display Character: ‐]]transcriptional level. The RNA-binding protein Qauking (QKI), originally known for its function in the nervous system, has been demonstrated to be essential for blood vessel formation. We find that QKI is highly expressed in quiescent ECs, in vitro and in vivo. In contrast, human umbilical vein ECs displayed reduced levels of QKI in response to the inflammatory stimuli TNF-α as well as in cells lacking cell-cell contacts, suggesting that QKI may act to enhance barrier function. To test this, we specifically abrogated QKI expression in ECs and measured their capacity to form a high-resistance monolayer with Electrical Cell-substrate Impedance Sensing. Silencing of QKI did not affect EC adhesion or spreading, but markedly affected the capacity to form a high resistance endothelial monolayer. Consistent with these data, and the fact that VE-cadherin mRNA contains a putative QKI-response element, the targeted reduction in QKI was accompanied by a significant reduction in VE-cadherin expression at cell junctions. Importantly, we identified a direct role for QKI in regulating VE-cadherin mRNA biology, as RNA immunoprecipitation and luciferase-reporter assays revealed that QKI can directly bind to the VE-cadherin mRNA and regulate transcript stability, respectively. In conclusion, we show that the modulation of QKI expression levels affects endothelial monolayer integrity by functioning as a critical regulator of the VE-cadherin mRNA. These studies provide novel insight into a role for post-transcriptional regulation in the maintenance of endothelial barrier function, and may have wide ranging implications for the preservation of vascular integrity in disease.
- © 2013 by American Heart Association, Inc.