Zhang Shichao, Zhou Peirou, Zhu Fanfan, Wang Yingying, Wang Xuesong, Chen Jingwen, Li Yumeng, Shao Xiaoyi
Department of Immunology, Medical School, Nantong University, Nantong 226019, China.
Biology (Basel). 2025 Aug 7;14(8):1017. doi: 10.3390/biology14081017.
We aimed to explore the mechanism by which extracellular-5'-nucleotidase (NT5E) regulates macrophage polarization via regenerating islet-derived protein 3 beta (Reg3β) and other plasma proteins that mediate immune-cell effects on myocarditis. Methods: The involvement of NT5E in Reg3β-induced macrophage polarization was first analyzed using RNA sequencing, Western blotting, and quantitative polymerase chain reaction. Mendelian randomization was employed to identify NT5E and various plasma proteins as potential therapeutic targets for myocarditis. Mediation analysis, enrichment analysis, protein-protein interaction network analysis, drug prediction, molecular docking, and single-cell RNA sequencing were integrated to further evaluate the biological functions and pharmacological potential of the identified targets. Finally, phenome-wide association studies were conducted to assess the safety of targeting these proteins. NT5E expression was elevated in Reg3β-stimulated M2 macrophages. The expression of Arg-1, a marker of M2 macrophages, decreased upon NT5E knockdown, suggesting that NT5E is involved in the Reg3β-mediated polarization of macrophages to the M2 phenotype. Mendelian randomization analysis identified NT5E and 80 other plasma proteins as being causally associated with myocarditis. Mediation analysis revealed 12 immune-cell types were mediators of the effects of plasma protein on myocarditis progression. Drug prediction identified candidates such as ICN 1229 and chrysin, which showed strong binding affinities in molecular docking analyses. These findings may contribute to the development of effective treatments for myocarditis. : NT5E plays a dual role in Reg3β-induced macrophage polarization and in interacting with plasma proteins that influence the onset and progression of myocarditis through immune-cell pathways.
我们旨在探索细胞外5'-核苷酸酶（NT5E）通过再生胰岛衍生蛋白3β（Reg3β）和其他介导免疫细胞对心肌炎影响的血浆蛋白来调节巨噬细胞极化的机制。方法：首先使用RNA测序、蛋白质印迹和定量聚合酶链反应分析NT5E在Reg3β诱导的巨噬细胞极化中的作用。采用孟德尔随机化方法确定NT5E和各种血浆蛋白作为心肌炎的潜在治疗靶点。综合中介分析、富集分析、蛋白质-蛋白质相互作用网络分析、药物预测、分子对接和单细胞RNA测序，进一步评估所确定靶点的生物学功能和药理潜力。最后，进行全表型关联研究以评估靶向这些蛋白的安全性。NT5E在Reg3β刺激的M2巨噬细胞中表达升高。NT5E敲低后，M2巨噬细胞标志物Arg-1的表达降低，表明NT5E参与Reg3β介导的巨噬细胞向M2表型的极化。孟德尔随机化分析确定NT5E和其他80种血浆蛋白与心肌炎存在因果关联。中介分析揭示12种免疫细胞类型是血浆蛋白对心肌炎进展影响的介导因子。药物预测确定了ICN 1229和白杨素等候选药物，它们在分子对接分析中显示出很强的结合亲和力。这些发现可能有助于开发有效的心肌炎治疗方法。：NT5E在Reg3β诱导的巨噬细胞极化以及与通过免疫细胞途径影响心肌炎发生和进展的血浆蛋白相互作用中发挥双重作用。
