Biogenesis of circRBM33 mediated by N6-methyladenosine and its function in abdominal aortic aneurysm.

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Epigenetics Pub Date : 2024-12-01 Epub Date: 2024-09-09 DOI:10.1080/15592294.2024.2392401
Yingqi Xu, Xiang Weng, Jiacong Qiu, Shizhi Wang
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引用次数: 0

Abstract

This study aimed to explore whether m6A modification affects the biogenesis of circRBM33, which is involved in the progression of abdominal aortic aneurysm (AAA). For in vitro experiments, vascular smooth muscle cells (VSMCs) were treated with Ang II. MeRIP‒PCR was used to assess m6A modification of circRBM33. Gene expression was measured using RT‒qPCR and Western blotting. For in vivo experiments, a mouse model of AAA was established via Ang II infusion. HE, Sirius Red and TUNEL staining was performed to evaluate pathological changes and cell apoptosis in aortic vessels. The results showed that the m6A level of circRBM33 was abnormally increased in Ang II-induced VSMCs. In addition, METTL3 positively regulated circRBM33 expression. YTHDC1 deficiency decreased circRBM33 expression but had no effect on RBM33 mRNA expression. Notably, neither METTL3 nor YTHDC1 influenced the stability of circRBM33 or RBM33 mRNA. The interaction between circRBM33 and METTL3/YTHDC1 was verified by RIP analysis. Moreover, the Ang II-induced increase in circRBM33 expression was reversed by cycloleucine (an inhibitor of m6A methylation). Importantly, the m6A modification and expression of circRBM33 in the circRBM33-m6A-mut2-expressing VSMCs were not altered by METTL3 silencing. Mechanistically, METTL3/YTHDC1 modulates the biogenesis of circRBM33 in an m6A-dependent manner. In addition, circRBM33 knockdown alleviated AAA by reducing ECM degradation in the Ang II-infused mice. In conclusion, this study demonstrated that METTL3/YTHDC1-mediated m6A modification modulates the biogenesis of circRBM33 from exons of the RBM33 gene. Moreover, knockdown of circRBM33 alleviated AAA by reducing ECM degradation, which may provide a novel therapeutic strategy for treating AAA.

由 N6-甲基腺苷介导的 circRBM33 的生物生成及其在腹主动脉瘤中的功能。
本研究旨在探讨 m6A 修饰是否会影响 circRBM33 的生物生成,而 circRBM33 与腹主动脉瘤(AAA)的进展有关。在体外实验中,用 Ang II 处理血管平滑肌细胞(VSMC)。MeRIP-PCR 用于评估 circRBM33 的 m6A 修饰。使用 RT-qPCR 和 Western 印迹法测量基因表达。在体内实验中,通过 Ang II 输注建立了 AAA 小鼠模型。用 HE、天狼星红和 TUNEL 染色法评估主动脉血管的病理变化和细胞凋亡。结果显示,在 Ang II 诱导的 VSMCs 中,circRBM33 的 m6A 水平异常升高。此外,METTL3 能正向调节 circRBM33 的表达。YTHDC1 缺乏会降低 circRBM33 的表达,但对 RBM33 mRNA 的表达没有影响。值得注意的是,METTL3 和 YTHDC1 都不会影响 circRBM33 或 RBM33 mRNA 的稳定性。circRBM33 与 METTL3/YTHDC1 之间的相互作用通过 RIP 分析得到了验证。此外,环亮氨酸(m6A 甲基化抑制剂)可逆转 Ang II 诱导的 circRBM33 表达增加。重要的是,在表达 circRBM33-m6A-mut2- 的 VSMCs 中,m6A 修饰和 circRBM33 的表达不会因 METTL3 沉默而改变。从机制上讲,METTL3/YTHDC1 以 m6A 依赖性方式调节 circRBM33 的生物生成。此外,circRBM33 基因敲除可减少血管紧张素 II 注入小鼠体内 ECM 的降解,从而缓解 AAA。总之,本研究证明了 METTL3/YTHDC1 介导的 m6A 修饰调节了来自 RBM33 基因外显子的 circRBM33 的生物生成。此外,敲除 circRBM33 可通过减少 ECM 降解缓解 AAA,这可能为治疗 AAA 提供了一种新的治疗策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Epigenetics
Epigenetics 生物-生化与分子生物学
CiteScore
6.80
自引率
2.70%
发文量
82
审稿时长
3-8 weeks
期刊介绍: Epigenetics publishes peer-reviewed original research and review articles that provide an unprecedented forum where epigenetic mechanisms and their role in diverse biological processes can be revealed, shared, and discussed. Epigenetics research studies heritable changes in gene expression caused by mechanisms others than the modification of the DNA sequence. Epigenetics therefore plays critical roles in a variety of biological systems, diseases, and disciplines. Topics of interest include (but are not limited to): DNA methylation Nucleosome positioning and modification Gene silencing Imprinting Nuclear reprogramming Chromatin remodeling Non-coding RNA Non-histone chromosomal elements Dosage compensation Nuclear organization Epigenetic therapy and diagnostics Nutrition and environmental epigenetics Cancer epigenetics Neuroepigenetics
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