PMID- 36254231 OWN - NLM STAT- MEDLINE DCOM- 20221021 LR - 20221021 IS - 1942-0994 (Electronic) IS - 1942-0900 (Print) IS - 1942-0994 (Linking) VI - 2022 DP - 2022 TI - CircCRIM1 Ameliorates Endothelial Cell Angiogenesis in Aging through the miR-455-3p/Twist1/VEGFR2 Signaling Axis. PG - 2062885 LID - 10.1155/2022/2062885 [doi] LID - 2062885 AB - BACKGROUND: Aging leads to vascular endothelial cell senescence. Decreased expression of VEGFA and VEGFR2 plays a crucial role in impairing angiogenesis in senescent endothelial cells. Noncoding RNAs, including circular RNAs (circRNAs) and microRNAs (miRNAs), regulate endothelial cell proliferation, differentiation, apoptosis, and migration and participate in the occurrence and development of vascular diseases. However, the mechanism of noncoding RNAs in age-related vascular endothelial dysfunction remains unclear. Here, we aimed to identify the circRNA that is associated with VEGF/VEGFR2 signaling pathway activation in angiogenesis. METHODS: Immunoblotting, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), in vitro and in vivo experiments, luciferase assays, and chromatin immunoprecipitation followed by qRT-PCR (ChIP-qPCR) assays were performed to clarify the roles played by circCRIM1 in mouse aortic endothelial cell (MAEC) angiogenesis. RESULTS: CircCRIM1 expression was downregulated in both an aging mouse model of lower limb ischemia in vivo and aging MAECs in vitro. Overexpressing circCRIM1 mediated through a plasmid or adeno-associated virus (AAV) reversed the downregulation of angiogenesis-related phenotype acquisition during aging. MiR-455-3p was confirmed to be a potential target of circCRIM1 through luciferase assays followed by RNA fluorescence in situ hybridization (FISH), which revealed the colocalization of circCRIM1 and miR-455-3p. CircCRIM1 was found to be a competitive endogenous RNA that sponged miR-455-3p and regulated angiogenesis-related phenotypes in MAECs. Furthermore, Twist1 was found to be downstream of miR-455-3p. A ChIP-qPCR assay showed that Twist1 promoted VEGFR2 expression by binding to the promoter region, playing a vital role in angiogenesis. CONCLUSIONS: Decreased expression of circCRIM1 impaired angiogenesis in aging via the miR-455-3p/Twist1/VEGFR2 axis. Our findings suggest that overexpression of circCRIM1 may be an effective therapeutic strategy for promoting ischemic lower limb blood flow recovery. CI - Copyright (c) 2022 Lei Zhao et al. FAU - Zhao, Lei AU - Zhao L AUID- ORCID: 0000-0003-1929-1744 AD - Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - Guangdong Engineering and Technology Center for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Chen, Rencong AU - Chen R AUID- ORCID: 0000-0001-8120-7289 AD - Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - Guangdong Engineering and Technology Center for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Qiu, Jiacong AU - Qiu J AD - Department of vascular Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang University, Nanchang, Jiangxi 330006, China. FAU - Huang, Yingxiong AU - Huang Y AD - Department of Emergency, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Lian, Chong AU - Lian C AD - Department of Endovascular Surgery, The First Affiliated Hospital of Zhengzhou University, 450000 Zhengzhou, Henan, China. FAU - Zhu, Xiaonan AU - Zhu X AD - Department of Pharmacology Laboratory, Zhongshan School of Medicine, Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Cui, Jin AU - Cui J AD - Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - Guangdong Engineering and Technology Center for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Wang, Siwen AU - Wang S AD - Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - Guangdong Engineering and Technology Center for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Wang, Shenming AU - Wang S AD - Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - Guangdong Engineering and Technology Center for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Hu, Zuojun AU - Hu Z AD - Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. AD - Guangdong Engineering and Technology Center for Diagnosis and Treatment of Vascular Diseases, NO. 58, Zhong Shan Er Lu, Guangzhou 510080, China. FAU - Wang, Jinsong AU - Wang J AUID- ORCID: 0000-0002-6829-6114 AD - Division of Vascular and Plastic Surgery, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China. LA - eng PT - Journal Article DEP - 20221008 PL - United States TA - Oxid Med Cell Longev JT - Oxidative medicine and cellular longevity JID - 101479826 RN - 0 (MicroRNAs) RN - 0 (RNA, Circular) RN - 0 (Vascular Endothelial Growth Factor A) RN - 0 (MIRN455 microRNA, mouse) RN - 136253-27-5 (Twist1 protein, mouse) RN - EC 2.7.10.1 (Kdr protein, mouse) RN - EC 2.7.10.1 (Vascular Endothelial Growth Factor Receptor-2) RN - 0 (Twist-Related Protein 1) SB - IM MH - Animals MH - Mice MH - Aging/genetics MH - Cell Proliferation/genetics MH - Endothelial Cells/metabolism MH - Gene Expression Regulation, Neoplastic MH - In Situ Hybridization, Fluorescence MH - *MicroRNAs/genetics MH - Neovascularization, Pathologic/genetics/metabolism MH - *RNA, Circular/genetics MH - Signal Transduction MH - Vascular Endothelial Growth Factor A/metabolism MH - *Vascular Endothelial Growth Factor Receptor-2/genetics/metabolism MH - *Twist-Related Protein 1/genetics/metabolism PMC - PMC9569221 COIS- The authors declare that there are no conflicts of interest. EDAT- 2022/10/19 06:00 MHDA- 2022/10/20 06:00 PMCR- 2022/10/08 CRDT- 2022/10/18 01:47 PHST- 2022/06/29 00:00 [received] PHST- 2022/09/22 00:00 [accepted] PHST- 2022/10/18 01:47 [entrez] PHST- 2022/10/19 06:00 [pubmed] PHST- 2022/10/20 06:00 [medline] PHST- 2022/10/08 00:00 [pmc-release] AID - 10.1155/2022/2062885 [doi] PST - epublish SO - Oxid Med Cell Longev. 2022 Oct 8;2022:2062885. doi: 10.1155/2022/2062885. eCollection 2022.