Zhou Kai, Zhou Jiaxiang, Luo XianJin, Chen Yan, Ao Jian, Wu Wei, Yang Bo, He Zhongyuan
Department of Orthopedics, People's Hospital of Chongqing Liang Jiang New Area, Chongqing, China.
Deparment of Orthopaedics, the Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, China.
Front Immunol. 2025 Jul 29;16:1606905. doi: 10.3389/fimmu.2025.1606905. eCollection 2025.
To investigate the role of hypoxia-related genes and immune infiltration in intervertebral disc degeneration (IDD) to identify molecular mechanisms and potential therapeutic targets.
Using GEO data, IDD-related gene expression datasets were analyzed for hypoxia-related differentially expressed genes (HRDEGs). Logistic regression and receiver operating characteristic (ROC) analyses were employed to evaluate the diagnostic potential of HRDEGs. Consensus clustering further delineated molecular subtypes of IDD. Functional enrichment analyses (GO, KEGG, GSEA) highlighted key pathways. Protein-protein interaction (PPI) networks were built in STRING and visualized with Cytoscape, identifying core genes with MCODE and CytoHubba. Immune cell infiltration was analyzed with CIBERSORT and ssGSEA to correlate immune cells with hypoxia-related genes. To validate the expression of potential biomarkers, qPCR and immunohistochemistry were conducted on human intervertebral disc tissue samples.
The integration of GSE150408 and GSE124272 datasets with batch effect removal enabled differential gene analysis, identifying nine HRDEGs, including RCOR2, STAT3, and NOTCH1. Logistic regression analysis demonstrated that these genes have high diagnostic efficacy for IDD. Co-expression and clustering analyses revealed two distinct molecular subtypes in IDD, each characterized by unique gene expression and immune infiltration profiles. Functional and pathway enrichment analyses also showed that these DEGs are involved in pathways regulating TP53 transcription, oxidative phosphorylation, and MAPK signaling, contributing to IDD pathology.
Hypoxia-related genes, particularly RCOR2, STAT3, and NOTCH1, play a significant role in the pathology of IDD and may serve as valuable diagnostic biomarkers and therapeutic targets, with distinct immune infiltration patterns associated with different IDD subtypes.
探讨缺氧相关基因和免疫浸润在椎间盘退变（IDD）中的作用，以确定分子机制和潜在治疗靶点。
利用基因表达综合数据库（GEO）数据，分析与IDD相关的基因表达数据集，以寻找缺氧相关差异表达基因（HRDEGs）。采用逻辑回归和受试者工作特征（ROC）分析来评估HRDEGs的诊断潜力。共识聚类进一步划分了IDD的分子亚型。功能富集分析（基因本体论、京都基因与基因组百科全书、基因集富集分析）突出了关键通路。在STRING中构建蛋白质-蛋白质相互作用（PPI）网络，并使用Cytoscape进行可视化，通过分子复合物检测（MCODE）和CytoHubba识别核心基因。使用CIBERSORT和单样本基因集富集分析（ssGSEA）分析免疫细胞浸润情况，以关联免疫细胞与缺氧相关基因。为验证潜在生物标志物的表达，对人椎间盘组织样本进行了定量聚合酶链反应（qPCR）和免疫组织化学检测。
去除批次效应后，整合GSE150408和GSE124272数据集进行差异基因分析，鉴定出9个HRDEGs，包括RCOR2、信号转导和转录激活因子3（STAT3）和Notch信号通路1（NOTCH1）。逻辑回归分析表明，这些基因对IDD具有较高的诊断效能。共表达和聚类分析揭示了IDD中的两种不同分子亚型，每种亚型都具有独特的基因表达和免疫浸润特征。功能和通路富集分析还表明，这些差异表达基因参与调节TP53转录、氧化磷酸化和丝裂原活化蛋白激酶（MAPK）信号通路，促进了IDD的病理过程。
缺氧相关基因，特别是RCOR2、STAT3和NOTCH1，在IDD病理过程中起重要作用，可能作为有价值的诊断生物标志物和治疗靶点，不同的IDD亚型具有独特的免疫浸润模式。
