Wu Yihao, Jiang Deying, Zhang Hao, Yin Fanxing, Guo Panpan, Zhang Xiaoxu, Bian Ce, Chen Chen, Li Shuixin, Yin Yuhan, Böckler Dittmar, Zhang Jian, Han Yanshuo
School of Life and Pharmaceutical Sciences, Dalian University of Technology, Panjin, China.
Department of Vascular Surgery, Dalian Municipal Central Hospital, Dalian, China.
Front Cardiovasc Med. 2022 May 10;9:883155. doi: 10.3389/fcvm.2022.883155. eCollection 2022.
This study aimed to identify key AAA-related m1A RNA methylation regulators and their association with immune infiltration in AAA. Furthermore, we aimed to explore the mechanism that m1A regulators modulate the functions of certain immune cells as well as the downstream target genes, participating in the progression of AAA.
Based on the gene expression profiles of the GSE47472 and GSE98278 datasets, differential expression analysis focusing on m1A regulators was performed on the combined dataset to identify differentially expressed m1A regulatory genes (DEMRGs). Additionally, CIBERSORT tool was utilized in the analysis of the immune infiltration landscape and its correlation with DEMRGs. Moreover, we validated the expression levels of DEMRGs in human AAA tissues by real-time quantitative PCR (RT-qPCR). Immunofluorescence (IF) staining was also applied in the validation of cellular localization of in AAA tissues. Furthermore, we established LPS/IFN-γ induced M1 macrophages and knockdown macrophages , to explore the relationship between and macrophage polarization. At last, RNA immunoprecipitation-sequencing (RIP-Seq) combined with PPI network analysis was used to predict the target genes of in AAA progression.
Eight DEMRGs were identified in our study, including as up-regulated genes and as down-regulated genes. The immune infiltration analysis showed these DEMRGs were positively correlated with activated mast cells, plasma cells and M1 macrophages in AAA. RT-qPCR analysis also verified the up-regulated expression levels of , and in human AAA tissues. Besides, IF staining result in AAA adventitia indicated the localization of in macrophages. Moreover, our experiments found that the knockdown of in M0 macrophages inhibits macrophage M1 polarization but promotes macrophage M2 polarization. Eventually, 30 key AAA-related target genes of were predicted, including , etc.
Our study reveals that m1A regulation is significantly associated with the pathogenesis of human AAA. The m1A "reader," , may participate in the modulating of macrophage polarization that promotes aortic inflammation, and influence AAA progression by regulating the expression of its target genes.
本研究旨在鉴定与腹主动脉瘤（AAA）相关的关键m1A RNA甲基化调节因子及其与AAA中免疫浸润的关联。此外，我们旨在探究m1A调节因子调节某些免疫细胞功能以及下游靶基因的机制，参与AAA的进展。
基于GSE47472和GSE98278数据集的基因表达谱，对合并后的数据集进行聚焦于m1A调节因子的差异表达分析，以鉴定差异表达的m1A调节基因（DEMRGs）。此外，使用CIBERSORT工具分析免疫浸润格局及其与DEMRGs的相关性。此外，我们通过实时定量PCR（RT-qPCR）验证了人AAA组织中DEMRGs的表达水平。免疫荧光（IF）染色也用于验证AAA组织中[具体蛋白名称未给出]的细胞定位。此外，我们建立了脂多糖/干扰素-γ诱导的M1巨噬细胞和[具体基因名称未给出]敲低的巨噬细胞，以探究[具体基因名称未给出]与巨噬细胞极化之间的关系。最后，RNA免疫沉淀测序（RIP-Seq）结合蛋白质-蛋白质相互作用（PPI）网络分析用于预测[具体基因名称未给出]在AAA进展中的靶基因。
我们的研究鉴定出8个DEMRGs，包括[上调基因名称未给出]作为上调基因和[下调基因名称未给出]作为下调基因。免疫浸润分析表明，这些DEMRGs与AAA中活化的肥大细胞、浆细胞和M1巨噬细胞呈正相关。RT-qPCR分析也验证了[具体基因名称未给出]、[具体基因名称未给出]和[具体基因名称未给出]在人AAA组织中的上调表达水平。此外，AAA外膜的IF染色结果表明[具体蛋白名称未给出]在巨噬细胞中的定位。此外，我们的[具体实验名称未给出]实验发现，M0巨噬细胞中[具体基因名称未给出]的敲低抑制巨噬细胞M1极化，但促进巨噬细胞M2极化。最终，预测了[具体基因名称未给出]的30个关键AAA相关靶基因，包括[具体基因名称未给出]等。
我们的研究表明，m1A调节与人类AAA的发病机制显著相关。m1A“读取器”[具体蛋白名称未给出]可能参与调节促进主动脉炎症的巨噬细胞极化，并通过调节其靶基因的表达影响AAA进展。
