Deficiency of protein inhibitor of activated STAT3 exacerbates atherosclerosis by modulating VSMC phenotypic switching

Haole Liu; Jingyi Zhang; Ziyang Xue; Mingke Chang; Xinxin Feng; Yifan Cai; Liang Bai; Weirong Wang; Enqi Liu; Sihai Zhao; Rong Wang

doi:10.1016/j.atherosclerosis.2023.117195

Deficiency of protein inhibitor of activated STAT3 exacerbates atherosclerosis by modulating VSMC phenotypic switching

Atherosclerosis. 2023 Sep:380:117195. doi: 10.1016/j.atherosclerosis.2023.117195. Epub 2023 Jul 27.

Authors

Haole Liu¹, Jingyi Zhang¹, Ziyang Xue¹, Mingke Chang¹, Xinxin Feng¹, Yifan Cai¹, Liang Bai², Weirong Wang², Enqi Liu², Sihai Zhao³, Rong Wang⁴

Affiliations

¹ Institute of Cardiovascular Science, Translational Medicine Institute, Xi'an Jiaotong University Health Science Center, Xi'an, 710061, China; Department of Laboratory Animal Science, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China.
² Institute of Cardiovascular Science, Translational Medicine Institute, Xi'an Jiaotong University Health Science Center, Xi'an, 710061, China; Laboratory Animal Center, Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China.
³ Laboratory Animal Center, Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China. Electronic address: sihaizhao@xjtu.edu.cn.
⁴ Institute of Cardiovascular Science, Translational Medicine Institute, Xi'an Jiaotong University Health Science Center, Xi'an, 710061, China; Department of Laboratory Animal Science, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China. Electronic address: rongw1986@mail.xjtu.edu.cn.

PMID: 37586220
DOI: 10.1016/j.atherosclerosis.2023.117195

Abstract

Background and aims: Phenotypic switching of vascular smooth muscle cells (VSMCs) plays an essential role in the development of atherosclerosis. Protein inhibitor of activated STAT (Pias) regulates VSMCs phenotype via acting as sumo E3 ligase to promote protein sumoylation. Our previous study indicated that Pias3 expression decreased in atherosclerotic lesions. Therefore, this study aimed to explore the role of Pias3 on VSMCs phenotype switching during atherosclerosis.

Methods: ApoE^-/- and ApoE^-/-Pias3^-/- double-deficient mice were fed with high-fat/high-cholesterol diet to induce atherosclerosis. Aorta tissues and primary VSMCs were collected to assess plaque formation and VSMCs phenotype. In vitro, Pias3 was overexpressed in A7r5, a VSMCs cell line, by transfection with Pias3 plasmid. Real-time quantitative PCR, immunoblotting, immunoprecipitation, were used to analyze the effect of Pias3 on VSMCs phenotypic switching.

Results: Pias3 deficiency significantly exacerbated atherosclerotic plaque formation and promoted VSMCs phenotypic switching to a synthetic state within lesion. In vitro, overexpressing Pias3 in VSMCs increased the expression of contractile markers (myosin heavy chain 11, calponin 1), while it decreased the level of synthetic marker (vimentin). Additionally, Pias3 overexpression blocked PDGF-BB-induced VSMCs proliferation and migration. Immunoprecipitation and mass spectrometry results showed that Pias3 enhanced sumoylation and ubiquitination of vimentin, and shortened its half-life. Moreover, the ubiquitination level of vimentin was impaired by 2-D08, a sumoylation inhibitor. This suggests that Pias3 might accelerate the ubiquitination-degradation of vimentin by promoting its sumoylation.

Conclusions: These results indicate that Pias3 might ameliorate atherosclerosis progression by suppressing VSMCs phenotypic switching and reducing vimentin protein stability.

Keywords: Atherosclerosis; Protein inhibitor of activated STAT3 (Pias3); Sumoylation; VSMCs phenotypic switching; Vascular smooth muscle cells (VSMCs).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apolipoproteins E / genetics
Atherosclerosis* / pathology
Cell Proliferation
Cells, Cultured
Mice
Muscle, Smooth, Vascular* / pathology
Myocytes, Smooth Muscle / pathology
Phenotype
Vimentin / genetics
Vimentin / metabolism

Substances

Vimentin
Apolipoproteins E