Oxido-reductive regulation of vascular remodeling by receptor tyrosine kinase ROS1
Ziad A. Ali, … , Thomas Quertermous, Euan A. Ashley
Ziad A. Ali, … , Thomas Quertermous, Euan A. Ashley
Published November 17, 2014
Citation Information: J Clin Invest. 2014;124(12):5159-5174. https://doi.org/10.1172/JCI77484.
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Research Article

Oxido-reductive regulation of vascular remodeling by receptor tyrosine kinase ROS1

Abstract

Angioplasty and stenting is the primary treatment for flow-limiting atherosclerosis; however, this strategy is limited by pathological vascular remodeling. Using a systems approach, we identified a role for the network hub gene glutathione peroxidase-1 (GPX1) in pathological remodeling following human blood vessel stenting. Constitutive deletion of Gpx1 in atherosclerotic mice recapitulated this phenotype of increased vascular smooth muscle cell (VSMC) proliferation and plaque formation. In an independent patient cohort, gene variant pair analysis identified an interaction of GPX1 with the orphan protooncogene receptor tyrosine kinase ROS1. A meta-analysis of the only genome-wide association studies of human neointima-induced in-stent stenosis confirmed the association of the ROS1 variant with pathological remodeling. Decreased GPX1 expression in atherosclerotic mice led to reductive stress via a time-dependent increase in glutathione, corresponding to phosphorylation of the ROS1 kinase activation site Y2274. Loss of GPX1 function was associated with both oxidative and reductive stress, the latter driving ROS1 activity via s-glutathiolation of critical residues of the ROS1 tyrosine phosphatase SHP-2. ROS1 inhibition with crizotinib and deglutathiolation of SHP-2 abolished GPX1-mediated increases in VSMC proliferation while leaving endothelialization intact. Our results indicate that GPX1-dependent alterations in oxido-reductive stress promote ROS1 activation and mediate vascular remodeling.

Authors

Ziad A. Ali, Vinicio de Jesus Perez, Ke Yuan, Mark Orcholski, Stephen Pan, Wei Qi, Gaurav Chopra, Christopher Adams, Yoko Kojima, Nicholas J. Leeper, Xiumei Qu, Kathia Zaleta-Rivera, Kimihiko Kato, Yoshiji Yamada, Mitsutoshi Oguri, Allan Kuchinsky, Stanley L. Hazen, J. Wouter Jukema, Santhi K. Ganesh, Elizabeth G. Nabel, Keith Channon, Martin B. Leon, Alain Charest, Thomas Quertermous, Euan A. Ashley

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Figure 10

Increased in-stent stenosis in Gpx1–/– Apoe–/– mice is reversed by ROS1 inhibition.

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Increased in-stent stenosis in Gpx1–/– Apoe–/– mice is reversed by ROS1 ...
(A) Relative area of atherosclerosis in the aortic root was significantly increased in Gpx1–/– Apoe–/– mice, but addition of genistein to inhibit ROS1 abolished this increase, while crizotinib did not. Scale bar: 50 μm. (B) Gpx1–/– Apoe–/– mice 4 weeks following BAS developed significantly greater neointimal hyperplasia and in-stent stenosis compared with control Gpx1+/+ Apoe–/–. Addition of genistein or crizotinib decreased neointimal hyperplasia to levels observed with the mTOR inhibitor rapamycin. AdGRX1 decreased neointimal hyperplasia to levels similar to those seen with crizotinib in Gpx1–/– Apoe–/– vessels, but not Gpx1+/+ Apoe–/–, suggesting deglutathiolation of SHP-2 as the mechanism. Scale bar: 50 μm. Black arrowheads denote neointima. Although overall neointimal cell density was higher in Gpx1–/– Apoe–/– mice, there was no difference in cell density when expressed per unit area between groups; however, addition of genistein, crizotinib, or rapamycin significantly decreased cell density per unit area in both experimental groups. Treatment with AdGrx1 reduced cell density only in Gpx1–/– Apoe–/– mice. *P < 0.05 compared with control; †P < 0.05 compared with AdGFP. n = 8–15/group. (C) Assessment of reendothelialization revealed that the ROS1 inhibitor crizotinib did not impair endothelial regeneration 4 weeks following BAS, while rapamycin did in both Gpx1–/– Apoe–/– Tie2-LacZ+ mice and control Gpx1+/+ Apoe–/– Tie2-LacZ+ mice. *P < 0.05 versus crizotinib. n = 6–10 mice/group. Gray arrowheads denote neointima. Black arrowheads denote LacZ-positive endothelial cells. Scale bar: 50 μm.