He Yuchen, Zhang Hao, Yin Fanxing, Guo Panpan, Wang Shiyue, Wu Yihao, Xin Shijie, Han Yanshuo, Zhang Jian
Department of Vascular Surgery, The First Hospital of China Medical University, 110001 Shenyang, Liaoning, China.
School of Life and Pharmaceutical Sciences, Dalian University of Technology, 116001 Dalian, Liaoning, China.
Front Biosci (Landmark Ed). 2021 Nov 30;26(11):1147-1165. doi: 10.52586/5016.
: It remains largely unclear about the function of 5-methylcytosine (m5C) RNA modification in the context of abdominal aortic aneurysm (AAA). In this regard, the present work focused on investigating m5C RNA methylation and related modulator expression levels in AAA. : To this end, we quantified the m5C methylation levels in AAA tissues (n = 32) and normal aortic tissues (n = 12) to examine the mRNA m5C status and m5C modulator expression at mRNA and protein levels. Meanwhile, modulator localization within AAA tissue samples was detected by immunohistochemistry (IHC). Moreover, RNA immunoprecipitation-sequencing (RIP-seq) was also used to analyze the lncRNAs and mRNA binding to Aly/REF, as an m5C reader. : m5C expression markedly elevated in AAA in comparison with normal aortic samples in the AAA cases. The major 5-methylcytosine modulators, including , , and , which represented the major parameters related to the abnormal m5C modification level, were observed up-regulating in AAA tissues at both protein and mRNA levels. In addition, mRNA level remarkably related to expression, and they were co-expressed in the same cells in AAA group. Regarding the cellular location, Aly/REF was associated with inflammatory (CD45+, CD3+) infiltrates. Simultaneously, after screening for reads in AAA tissue compare with anti-Aly/REF group relative to IgG as control, we obtained totally 477 differentially expressed Aly/REF-binding lncRNAs and 369 differentially expressed Aly/REF-binding mRNAs in AAA tissue. The functions of Aly/REF-interacting lncRNA were involved in immune system process and macrophages infiltration. Through regulatory network (lncRNA-mRNA) analysis, our findings predicted the potential mechanism of Aly/REF-induced lncBCL2L1 and Aly/REF-lncFHL1 axis in AAA and inspire the understanding of m5C and lncRNA in AAA. : This study is the first to examine m5A modification within human AAA samples. Our results indicate that m5C modulators, namely, Aly/REF and NUSN2, play vital parts in the human AAA pathogenic mechanism, which shed new lights on the function of m5C modification within AAA. Taken together, findings in this work offer a possible RNA methylation modification mechanism within clinical AAA.
在腹主动脉瘤（AAA）的背景下，5-甲基胞嘧啶（m5C）RNA修饰的功能在很大程度上仍不清楚。在这方面，目前的工作重点是研究AAA中m5C RNA甲基化和相关调节因子的表达水平。为此，我们对AAA组织（n = 32）和正常主动脉组织（n = 12）中的m5C甲基化水平进行了定量，以检测mRNA的m5C状态以及m5C调节因子在mRNA和蛋白质水平的表达。同时，通过免疫组织化学（IHC）检测AAA组织样本中调节因子的定位。此外，RNA免疫沉淀测序（RIP-seq）也用于分析与作为m5C读取器的Aly/REF结合的lncRNA和mRNA。与正常主动脉样本相比，AAA中m5C表达明显升高。在AAA组织中，主要的5-甲基胞嘧啶调节因子，包括[具体因子1]、[具体因子2]和[具体因子3]，代表了与异常m5C修饰水平相关的主要参数在蛋白质和mRNA水平均上调。此外，[基因名称1]的mRNA水平与[基因名称2]的表达显著相关，并且它们在AAA组的同一细胞中共表达。关于细胞定位，Aly/REF与炎症浸润（CD45+、CD3+）相关。同时，在将AAA组织中的读数与以IgG为对照的抗Aly/REF组进行比较筛选后，我们在AAA组织中总共获得了477个差异表达的Aly/REF结合lncRNA和369个差异表达的Aly/REF结合mRNA。与Aly/REF相互作用的lncRNA的功能涉及免疫系统过程和巨噬细胞浸润。通过调控网络（lncRNA-mRNA）分析，我们的研究结果预测了Aly/REF诱导的lncBCL2L1和Aly/REF-lncFHL1轴在AAA中的潜在机制，并促进了对AAA中m5C和lncRNA的理解。这项研究是首次在人类AAA样本中检测m5A修饰。我们的结果表明，m5C调节因子，即Aly/REF和NUSN2在人类AAA致病机制中起着至关重要的作用，这为AAA中m5C修饰的功能提供了新的线索。综上所述，这项工作的发现为临床AAA提供了一种可能的RNA甲基化修饰机制。
