Feng Anlin, Liang Ying, Fu Panfeng, Dong Yishu, Black Stephen M, Wang Ting
Center for Translational Science, Florida International University, Port Saint Lucie, FL 34987, USA; Department of Environmental Health Sciences, Florida International University, Miami, FL 33199, USA.
Center for Translational Science, Florida International University, Port Saint Lucie, FL 34987, USA; Department of Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USA.
Biochim Biophys Acta Mol Basis Dis. 2025 Aug;1871(6):167907. doi: 10.1016/j.bbadis.2025.167907. Epub 2025 May 14.
Acute Lung Injury (ALI) involves diffuse alveolar damage, neutrophil infiltration, and pulmonary edema, with unacceptable mortality. Bacterial lipopolysaccharide (LPS) activates inflammatory pathways in ALI, which are then regulated by transcriptional and post-transcriptional pathways to affect gene expression. RNA methylation, N6-methyladenosine, is the main m6A mRNA modification that controls the expression of various genes in different environments. There are very few facts about LPS's effect on m6A RNA methylation. This study will explore the m6A RNA methylation landscape in lung endothelial cells (ECs) to understand its role in lung inflammation. In this study, lung endothelial cells were treated with LPS, and the dynamics of mRNA m6A methylation were examined through m6A-methylated RNA sequencing. RNA abundance was measured with RNA-seq, and global protein expression and m6A-binding proteins were identified using mass spectrometry (MS). Following LPS treatment, global m6A methylation levels increased along with the upregulation and nuclear translocation of METTL3 protein, while demethylase activity remained unchanged. METTL3 drove LPS-induced m6A methylation and endothelial injury, as shown by selective METTL3 siRNA and the inhibitor STM2457. MeRIP-seq analyses revealed increased m6A sites near the 5' UTR in LPS-treated cells, with m6A methylation correlating positively with gene expression. The metabolic and apoptosis pathways were shown to be more enriched in different types of methylated exons. METTL3-mediated m6A methylation targeted inflammatory genes, enhancing protein expression in chemokine signaling and MAPK pathways. STM2457 effectively mitigated LPS- or CLP-induced experimental ALI. According to this paper, LPS-mediated m6A RNA methylation is described in terms of genomic structure. Modulation of m6A methylation exerts influence over LPS-mediated endothelial gene expression and the ensuing inflammatory response.
急性肺损伤（ALI）涉及弥漫性肺泡损伤、中性粒细胞浸润和肺水肿，死亡率高。细菌脂多糖（LPS）激活ALI中的炎症信号通路，随后通过转录和转录后通路进行调控，从而影响基因表达。RNA甲基化，即N6-甲基腺苷，是主要的m6A mRNA修饰，可在不同环境中控制各种基因的表达。关于LPS对m6A RNA甲基化的影响，目前所知甚少。本研究将探索肺内皮细胞（ECs）中的m6A RNA甲基化图谱，以了解其在肺部炎症中的作用。在本研究中，用LPS处理肺内皮细胞，并通过m6A甲基化RNA测序检测mRNA m6A甲基化的动态变化。用RNA测序测量RNA丰度，并用质谱（MS）鉴定整体蛋白表达和m6A结合蛋白。LPS处理后，整体m6A甲基化水平随着METTL3蛋白的上调和核转位而增加，而脱甲基酶活性保持不变。选择性METTL3 siRNA和抑制剂STM2457表明，METTL3驱动LPS诱导的m6A甲基化和内皮损伤。MeRIP-seq分析显示，LPS处理的细胞中5'UTR附近的m6A位点增加，m6A甲基化与基因表达呈正相关。代谢和凋亡通路在不同类型的甲基化外显子中富集程度更高。METTL3介导的m6A甲基化靶向炎症基因，增强趋化因子信号传导和MAPK通路中的蛋白表达。STM2457有效减轻了LPS或CLP诱导的实验性ALI。根据本文，从基因组结构方面描述了LPS介导的m6A RNA甲基化。m6A甲基化的调节对LPS介导的内皮基因表达和随后的炎症反应产生影响。
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