Targeting deubiquitinase OTUB1 protects vascular smooth muscle cells in atherosclerosis by modulating PDGFRβ

Abstract

Atherosclerosis is a chronic artery disease that causes various types of cardiovascular dysfunction. Vascular smooth muscle cells (VSMCs), the main components of atherosclerotic plaque, switch from contractile to synthetic phenotypes during atherogenesis. Ubiquitylation is crucial in regulating VSMC phenotypes in atherosclerosis, and it can be reversely regulated by deubiquitinases. However, the specific effects of deubiquitinases on atherosclerosis have not been thoroughly elucidated. In this study, RNAi screening in human aortic smooth muscle cells was performed to explore the effects of OTU family deubiquitinases, which revealed that silencing OTUB1 inhibited PDGF-BB-stimulated VSMC phenotype switch. Further in vivo studies using Apoe^−/− mice revealed that knockdown of OTUB1 in VSMCs alleviated atherosclerosis plaque burden in the advanced stage and led to a stable plaque phenotype. Moreover, VSMC proliferation and migration upon PDGF-BB stimulation could be inhibited by silencing OTUB1 in vitro. Unbiased RNA-sequencing data indicated that knocking down OTUB1 influenced VSMC differentiation, adhesion, and proliferation. Mass spectrometry of ubiquitinated protein confirmed that proteins related to cell growth and migration were differentially ubiquitylated. Mechanistically, we found that OTUB1 recognized the K707 residue ubiquitylation of PDGFRβ with its catalytic triad, thereby reducing the K48-linked ubiquitylation of PDGFRβ. Inhibiting OTUB1 in VSMCs could promote PDGFRβ degradation via the ubiquitin–proteasome pathway, so it was beneficial in preventing VSMCs’ phenotype switch. These findings revealed that knocking down OTUB1 ameliorated VSMCs’ phenotype switch and atherosclerosis progression, indicating that OTUB1 could be a valuable translational therapeutic target in the future.

Keywords

atherosclerosis / vascular smooth muscle cell / ubiquitylation / deubiquitinase / OTUB1 / PDGFRβ

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Acknowledgements

The authors genuinely thank Professor Yongming Wang from Fudan University for his assistance in composing the manuscript. This work was supported by grants from the National Key R&D Program of China (No. 2021YFC2500500), the National Natural Science Foundation of China (Nos. T2288101 and 82170342), Shanghai Engineering Research Center of Interventional Medicine (No. 19DZ2250300), and Shanghai Clinical Research Center for Interventional Medicine (No. 19MC1910300).

Electronic Supplementary Material

Supplementary material is available in the online version of this article at https://doi.org/10.1007/s11684-024-1056-8 and is accessible for authorized users.

Compliance with ethics guidelines

Conflict of interests Fei Xu, Han Chen, Changyi Zhou, Tongtong Zang, Rui Wang, Shutong Shen, Chaofu Li, Yue Yu, Zhiqiang Pei, Li Shen, Juying Qian, and Junbo Ge declare no conflicts of interest.

All animal experiments were approved by the ethics committee at Zhongshan Hospital, Fudan University, and they were performed according to the local relevant guidelines. The experiments conformed to the guidelines from Directive 2010/63/EU of the European Parliament on the protection of animals used for scientific purposes. All institutional and national guidelines for the care and use of laboratory animals were followed.

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