■椎间盘退变(IDD)是下腰痛(LBP)的主要原因。IDD发展和进展的机制尚不完全清楚。外周生物标志物在IDD的早期检测和诊断中日益成为重要的非放射性方法。然而,线粒体自噬基因在IDD进展中的作用研究较少。本研究旨在鉴定IDD发生过程中的线粒体自噬致病基因和诊断IDD的线粒体自噬生物标志物。
■通过分析来自GEO的IDD病例的微阵列数据集,研究了与IDD相关的线粒体自噬相关差异表达基因(MRDEG)。PathCards和分子签名数据库。我们用了R软件,WGCNA,PPI,mRNA-miRNA,mRNA-TF,GO,KEGG,GSEA,GSVA和Cytoscape来分析和可视化数据。我们进一步使用ssGSEA进行免疫浸润分析,以获得不同的免疫细胞浸润。开发LASSO模型来筛选满足诊断基因模型要求的基因。最后,qRT-PCR,使用蛋白质印迹和HE来验证来自临床IDD样品的hub基因及其表达。
■我们鉴定了14个MRDEGs和12个hub基因。GO,KEGG,GSEA和GSVA分析表明,hub基因对IDD的发展至关重要。LASSO诊断模型由六个中心基因组成,其中SQSTM1,ATG7和OPTN在两种IDD疾病亚型之间存在显着差异。同时,SQSTM1与免疫特征亚型也有高度相关性。qRT-PCR和Western印迹的结果还表明,这些基因在IDD组的髓核细胞(NPCs)中明显差异表达。
■我们探索了IDD中MRDEG相关特征之间的关联,并验证了像SQSTM1这样的中枢基因可能作为IDD诊断和治疗靶标的生物标志物。同时,本研究为研究线粒体自噬在IDD发生发展过程中的功能特征和作用机制提供了新的思路。
UNASSIGNED: Intervertebral disc degeneration (IDD) is the leading cause of low back pain (LBP). The mechanism of IDD development and progression is not fully understood. Peripheral biomarkers are increasingly vital non-radioactive methods in early detection and diagnosis for IDD. Nevertheless, less attention has been paid to the role of mitophagy genes in the progress of IDD. This study aimed to identify the mitophagy disease-causing genes in the process of IDD and mitophagy diagnostic biomarkers for IDD.
UNASSIGNED: Mitophagy-related differentially expressed genes (MRDEGs) related to IDD were investigated by analyzing the microarray datasets of IDD cases from GEO, PathCards and Molecular Signatures Databases. We used R software, WGCNA, PPI, mRNA-miRNA, mRNA-TF, GO, KEGG, GSEA, GSVA and Cytoscape to analyze and visualize the data. We further used ssGSEA for immunoinfiltration analysis to obtain different immune cell infiltration. LASSO model was developed to screen for genes that met the diagnostic gene model requirements. Finally, qRT-PCR, Western blotting and HE were used to verify hub genes and their expression from clinical IDD samples.
UNASSIGNED: We identified 14 MRDEGs and 12 hub genes. GO, KEGG, GSEA and GSVA analyses demonstrated that hub genes were critical for the development of IDD. LASSO diagnostic model consisted of six hub genes, among which
SQSTM1, ATG7 and OPTN were significantly different between the two IDD disease subtypes. At the same time,
SQSTM1 also had a high correlation with immune characteristic subtypes. The results of qRT-PCR and Western blotting also indicated that these genes were significantly differentially expressed in nucleus pulposus cells (NPCs) of the IDD group.
UNASSIGNED: We explored an association between MRDEGs-associated signature in IDD and validated that hub genes like
SQSTM1 might serve as biomarkers for diagnostic and therapeutic targets for IDD. Meanwhile, this study can provide new insights into the functional characteristics and mechanism of mitophagy in the development of IDD.