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 4

ROS1 is upregulated in in-stent neointima and involved in basic VSMC fates.

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ROS1 is upregulated in in-stent neointima and involved in basic VSMC fat...
(A) Transcriptional profiling of human coronary artery atherectomy specimens of control (AHA class I lesions), de novo atherosclerosis (AHA class III–V lesions), or in-stent stenosis identified that ROS1 expression was increased in atherosclerosis and furthermore in in-stent stenosis. *P < 0.01 vs. control; #P < 0.05 vs. atherosclerosis. n = 55 atherosclerosis/34 in-stent stenosis. (B) mRNA levels of Ros1 in Gpx1+/+ Apoe–/– mice were dramatically higher in aorta that had undergone BAS compared with uninjured aorta harvested at 28 days. *P < 0.001. n = 4/group. (C) HCASMCs were grown under both serum-fed proliferative and serum-starved nonproliferative phenotypes. ROS1 expression was minimal, but was upregulated as these cells differentiated into “contractile” phenotype and further by confluence, returning to contractile phenotype levels following passage. ROS1 expression was significantly decreased by siROS1. *P < 0.05 vs. (+) serum; **P < 0.05 vs. confluent; †P < 0.05 vs. (–) serum. (D) In primary aortic VSMCs from experimental animals that underwent angioplasty, proliferation was higher in Gpx1–/– Apoe–/– VSMCs compared with control Gpx1+/+ Apoe–/– VSMCs. siROS1, genistein, and crizotinib decreased proliferation in Gpx1–/– Apoe–/– mice, but not in Gpx1+/+ Apoe–/– controls, indicating that the phenotype was dependent on GPX1 deficiency. Daidzein, the structural analogue of genistein with similar antioxidant effects but no effect on tyrosine kinase inhibition, did not inhibit proliferation in Gpx1–/– Apoe–/– cells. Addition of siROS1 to Daidzein showed a reduction in proliferation similar to that of siROS1 alone. *P < 0.05 vs. Gpx1+/+ Apoe–/–; †P < 0.05 vs. baseline Gpx1–/– Apoe–/–.