激素性股骨头坏死中m6A相关基因差异性的鉴定  

Identification of differences in N6-methyladenosine-related genes in steroid-induced femoral head necrosis

作　　者：原天祎 刘洪江 杨增强 卢兴宝 麦麦提依不巴吉 周治衡 崔泳[1,2] Yuan Tianyi;Liu Hongjiang;Yang Zengqiang;Lu Xingbao;Maimaitiyibubaji;Zhou Zhiheng;Cui Yong(The Fifth Clinical Medical College of Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Department of Orthopedics,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第五临床医学院,新疆维吾尔自治区乌鲁木齐市830011 [2]新疆医科大学第五附属医院骨科中心,新疆维吾尔自治区乌鲁木齐市830011

出　　处：《中国组织工程研究》2024年第14期2159-2165,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81960396),项目负责人:崔泳。

摘　　要：背景:已知m6A参与了多种疾病的发生与发展,研究推测其也参与了激素性股骨头坏死的病理改变,但m6A甲基化修饰在激素性股骨头坏死中的研究比较匮乏。目的:通过生物信息学的方法寻找m6A相关基因在激素性股骨头坏死中的差异性表达,并预测与其相关的miRNA,以进一步阐明m6A甲基化在激素性股骨头坏死中所发挥的作用和机制。方法:从GSE123568基因表达谱中分析激素性股骨头坏死与对照组差异基因的表达,通过R语言‘limma’包进行差异基因鉴定,并对差异基因进行功能富集分析。用R语言的‘ggstatsplot’包对相关基因进行差异性分析。通过GSE74089数据集对差异基因进行交叉验证。构建mRNA-miRNA调控网络。采用共表达分析对共表达模块进行聚类并对模块基因进行富集分析。采用ssGSEA方法对激素性股骨头坏死与对照组之间免疫细胞浸润的差异进行量化分析。结果与结论:①通过相关性分析发现13个m6A相关基因,进行PPI网络识别和受试者操作特征曲线进一步分析发现YTHDF2有望成为早期生物标志物的核心差异基因;②富集分析发现差异基因主要参与炎症反应和免疫应答,并与破骨细胞关系密切;③交叉验证分析2个数据集差异基因表达结果一致;④mRNA-miRNA调控网络分析发现YTHDF2与miRNA-27a呈负相关;⑤免疫浸润分析发现激素性股骨头坏死中免疫细胞浸润水平上升,YTHDF2与免疫细胞CD4^(+)T细胞的浸润程度呈正相关;⑥结论:m6A相关基因YTHDF2可以作为激素性股骨头坏死潜在生物标志物,对激素性股骨头坏死的早期临床诊断和治疗具有一定的价值;验证了YTHDF2与miRNA-27a呈负相关,且与CD4^(+)T细胞的浸润程度呈正相关,为激素性股骨头坏死的早期诊断和治疗提供新的思路,并为进一步阐明m6A在激素性股骨头坏死中发挥的作用机制提出新的见解。BACKGROUND:It is known that N6-methyladenosine(m6A)plays a role in the pathogenesis of various diseases and studies have suggested its involvement in the pathologic changes of steroid-induced femoral head necrosis(SNFH).However,research on m6A methylation modifications in steroid-induced femoral head necrosis is limited.OBJECTIVE:Using bioinformatics methods to identify the differential expression of m6A-related genes in steroid-induced femoral head necrosis and to predict miRNAs associated with these genes to further elucidate the role and mechanism of m6A methylation in steroid-induced femoral head necrosis.METHODS:Differential gene expression between steroid-induced femoral head necrosis and control groups was analyzed using GSE123568 gene expression data and identified using the“limma”package in R.Functional enrichment analysis was performed on the differentially expressed genes.Differential analysis of the related genes was carried out using the“ggstatsplot”package in R.The differential genes were cross-validated using the GSE74089 dataset.An mRNAmiRNA regulatory network was constructed,and co-expression analysis was performed on the module genes followed by enrichment analysis.Differences in immune cell infiltration between steroid-induced femoral head necrosis and control groups were quantified using the ssGSEA method.RESULTS AND CONCLUSION:Correlation analysis revealed 13 m6A-related genes,and further analysis through the protein-protein interaction network identification and receiver operating characteristic curve analysis showed that YTHDF2 was expected to be a core differential gene as a potential early biomarker.Enrichment analysis indicated that differentially expressed genes were mainly involved in inflammation and immune response and were closely related to osteoclasts.Cross-validation analysis showed that differential gene expression results between the two datasets were consistent.mRNA-miRNA regulatory network analysis revealed that YTHDF2 was negatively correlated with miRNA-27a.Immune i

关 键 词：m6A甲基化 激素性股骨头坏死 生信分析 差异基因 YTHDF2 富集分析 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R681