Methyltransferase 3-mediated m6A modification of Switch/sucrose non-fermenting-related matrix-associated actin-dependent regulator of chromatin subfamily a member 5 promotes mycobacterium tuberculosis-infected macrophage M1 polarization and inflammation.

IF 3.1 4区医学 Q2 PATHOLOGY

Cytojournal Pub Date : 2025-04-01 eCollection Date: 2025-01-01 DOI:10.25259/Cytojournal_144_2024

Cong Chen, Hai Huang

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Abstract

Objective: Mycobacterium tuberculosis (MTB) manipulates macrophage functions, thus mediating tuberculosis (TB) progression. Whether the switch/sucrose non-fermenting-related matrix-associated actin-dependent regulator of chromatin subfamily a member 5 (SMARCA5) mediates MTB-induced macrophage polarization remains unclear.

Material and methods: Human Promyelocytic Leukemia Cell Line was induced into macrophages and then treated with MTB. Cell viability and apoptosis were tested with cell counting kit 8 assay and flow cytometry. Classically activated macrophages (M1) polarization and inflammation were measured by detecting CD86⁺ cell rate and inflammatory factor levels. The levels of SMARCA5, methyltransferase 3 (METTL3), and insulin-like growth factor 2 binding protein 1 (IGF2BP1) were assessed using quantitative real-time polymerase chain reaction or Western blot. The interaction between SMARCA5 and METTL3 or IGF2BP1 was confirmed by methylated RNA immunoprecipitation (RIP) and RIP assays. The effect of METTL3 knockdown on SMARCA5 messenger RNA (mRNA) stability was evaluated using actinomycin D treatment.

Results: MTB treatment suppressed the viability and promoted the apoptosis and M1 polarization and inflammation of macrophages (P < 0.05), and SMARCA5 knockdown abolished these effects (P < 0.05). METTL3 mediated the m6A methylation of SMARCA5 to enhance the mRNA stability of the latter, and this modification was recognized by IGF2BP1. SMARCA5 upregulation reverted the si-METTL3-mediated inhibition of MTB-induced macrophage M1 polarization and inflammation (P < 0.05).

Conclusion: METTL3-mediated SMARCA5 facilitates macrophage M1 polarization and inflammation, providing a novel target for TB treatment.

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甲基转移酶3介导的开关/蔗糖非发酵相关基质相关肌动蛋白依赖调节因子染色质亚家族成员5的m6A修饰促进结核分枝杆菌感染的巨噬细胞M1极化和炎症。

目的：结核分枝杆菌（MTB）操纵巨噬细胞功能，从而介导结核病（TB）进展。开关/蔗糖非发酵相关基质相关肌动蛋白依赖性染色质亚家族成员5 （SMARCA5）是否介导mmb诱导的巨噬细胞极化尚不清楚。材料和方法：将人早幼粒细胞白血病细胞系诱导成巨噬细胞，然后用结核分枝杆菌治疗。采用细胞计数试剂盒8和流式细胞术检测细胞活力和凋亡。通过检测CD86+细胞率和炎症因子水平，检测经典活化巨噬细胞（M1）极化和炎症。采用实时定量聚合酶链反应或Western blot检测SMARCA5、甲基转移酶3 （METTL3）和胰岛素样生长因子2结合蛋白1 （IGF2BP1）的水平。通过甲基化RNA免疫沉淀（RIP）和RIP实验证实了SMARCA5与METTL3或IGF2BP1之间的相互作用。使用放线菌素D处理评估METTL3敲低对SMARCA5信使RNA （mRNA）稳定性的影响。结果：MTB治疗可抑制巨噬细胞活力，促进巨噬细胞凋亡、M1极化和炎症（P < 0.05），而敲低SMARCA5可消除这些作用（P < 0.05）。METTL3介导SMARCA5的m6A甲基化，增强后者mRNA的稳定性，这种修饰被IGF2BP1识别。SMARCA5上调逆转了si- mettl3介导的对mtb诱导的巨噬细胞M1极化和炎症的抑制（P < 0.05）。结论：mettl3介导的SMARCA5促进巨噬细胞M1极化和炎症，为结核病治疗提供了新的靶点。

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来源期刊

Cytojournal PATHOLOGY-

CiteScore

2.20

自引率

42.10%

发文量

审稿时长

>12 weeks

期刊介绍： The CytoJournal is an open-access peer-reviewed journal committed to publishing high-quality articles in the field of Diagnostic Cytopathology including Molecular aspects. The journal is owned by the Cytopathology Foundation and published by the Scientific Scholar.