The lncRNA Punisher Inhibits Apoptosis of Vascular Smooth Muscle Cells Through Regulating Mitochondrial Homeostasis via Targeting miR-664a-5p and OPA1

Background: Long noncoding RNAs (lncRNA) are important regulators of various cellular functions in development and diseases. Recent studies have shown that the lncRNA Punisher is highly expressed in cardiovascular progenitors and have potential role in cardiovascular diseases. However, its role, especially molecular mechanism, is unclear. Methods: We performed RNA microarray assays to identify targets of PUNISHER. We investigated the effect of PUNISHER on proliferation, invasion, apoptosis and mitochondrial fission by knocking down and overexpressing this lncRNA in VSMCs. The interactions of molecules were studies by the bioinformatic analyses, luciferase report assay, RNA pull down, and RNA immunoprecipitation. Finally, we carried out clinical evaluations in patients with atherosclerosis to evaluate the relevance of PUNISHER in this kind of disease. Findings: In our study, we observed that Punisher suppressed the apoptosis of VSMC which potentially contributed to the progression of atherosclerosis. Intriguingly, Punisher was proved to regulate mitochondria fission and fusion as well as mitochondrial functions in VSMC. Mechanistically, Punisher serves as a ceRNA to regulate downstream target genes. It directly binds to miR-664a-5p and consequently regulates its target OPA1, which regulates the biological function of VSMC. Particularly, these results were in accordance with findings obtained with the clinical evaluation of patients with atherosclerosis. Our data was the first time to investigate the close relationship among OPA1, mitochondrial homeostatis, VSMC apoptosis and atherosclerosis. Interpretation: lncRNA Punisher and miR-664a-5p could serve as the novel and potential targets in the diagnosis and therapeutics of cardiovascular disease. Funding Statement: This work was supported by National Natural Science Foundation of China (No. 81870331, 31701208), Shandong Province: Taishan Scholars Project (tsqn201812044), China Postdoctoral Science Foundation (No. 2017M612189, 2016M590616), Natural Science Foundation of Shandong Province (No. ZR2017MC067), and The People’s livelihood science and technology project of Qingdao (No. 18-2-2-65-jch). Declaration of Interests: The authors have declared that no competing interest exists. Ethics Approval Statement: The research was approved by the Institutional Review Boards of Qingdao University Health Science Center. Informed consent was obtained from all participants prior to study participation. The housing, care and all the experimental procedures were approved by the Animal Care Committee.