Zhang Yongbo, Chen Liuyang, Yang Sheng, Dai Rui, Sun Hua, Zhang Liang
Department of Orthopedics Northern Jiangsu People's Hospital Affiliated to Yangzhou University Yangzhou China.
Department of Orthopedics The Yangzhou School of Clinical Medicine of Dalian Medical University Yangzhou China.
JOR Spine. 2025 Apr 13;8(2):e70066. doi: 10.1002/jsp2.70066. eCollection 2025 Jun.
Low back pain is a significant burden worldwide, and intervertebral disc degeneration (IVDD) is identified as the primary cause. Recent research has emphasized the significant role of circadian rhythms (CRs) and immunity in affecting intervertebral discs (IVD). However, the influence of circadian rhythms and immunity on the mechanism of IVDD remains unclear. This study aimed to identify and validate key rhythm-related genes in IVDD and analyze their correlation with immune cell infiltration.
Two gene expression profiles related to IVDD and rhythm-related genes were obtained from the Gene Expression Omnibus and GeneCards databases to identify differentially expressed rhythm-related genes (DERGs). Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene set enrichment analysis (GSEA) were conducted to explore the biological functions of these genes. LASSO regression and SVM algorithms were employed to identify hub genes. We subsequently investigated the correlation between hub rhythm-related genes and immune cell infiltration. Finally, nucleus pulposus-derived mesenchymal stem cells (NPMSCs) were isolated from normal and degenerative human IVD tissues. Hub rhythm-related genes expression in NPMSCs was confirmed by real-time quantitative PCR (RT-qPCR).
Six hub genes related to CRs (CCND1, FOXO1, FRMD8, NTRK2, PRRT1, and TFPI) were screened out. Immune infiltration analysis revealed that the IVDD group had significantly more M0 macrophages and significantly fewer follicular helper T cells than those of the control group. Specifically, M0 macrophages were significantly associated with FRMD8, PRRT1, and TFPI. T follicular helper cells were significantly associated with FRDM8, FOXO1, and CCND1. We further confirmed that CCND1, FRMD8, NTRK2, and TFPI were dysrhythmic within NPMSCs from degenerated IVD in vitro.
Six genes (CCND1, FOXO1, FRMD8, NTRK2, PRRT1 and TFPI) linked to circadian rhythms associated with IVDD progression, together with immunity. The identification of these DEGs may provide new insights for the diagnosis and treatment of IVDD.
腰痛是全球范围内的一项重大负担，而椎间盘退变（IVDD）被认为是主要原因。最近的研究强调了昼夜节律（CRs）和免疫在影响椎间盘（IVD）方面的重要作用。然而，昼夜节律和免疫对IVDD机制的影响仍不清楚。本研究旨在识别和验证IVDD中与节律相关的关键基因，并分析它们与免疫细胞浸润的相关性。
从基因表达综合数据库（Gene Expression Omnibus）和基因卡片数据库（GeneCards）中获取与IVDD和节律相关基因的两个基因表达谱，以识别差异表达的节律相关基因（DERGs）。进行基因本体论（GO）、京都基因与基因组百科全书（KEGG）和基因集富集分析（GSEA）以探索这些基因的生物学功能。采用套索回归和支持向量机算法识别枢纽基因。随后，我们研究了枢纽节律相关基因与免疫细胞浸润之间的相关性。最后，从正常和退变的人类IVD组织中分离出髓核来源的间充质干细胞（NPMSCs）。通过实时定量PCR（RT-qPCR）确认NPMSCs中枢纽节律相关基因的表达。
筛选出六个与CRs相关的枢纽基因（CCND1、FOXO1、FRMD8、NTRK2、PRRT1和TFPI）。免疫浸润分析显示，与对照组相比，IVDD组的M0巨噬细胞明显更多，滤泡辅助性T细胞明显更少。具体而言，M0巨噬细胞与FRMD8、PRRT1和TFPI显著相关。滤泡辅助性T细胞与FRDM8、FOXO1和CCND1显著相关。我们进一步证实，CCND1、FRMD8、NTRK2和TFPI在体外退变IVD的NPMSCs中节律异常。
六个与昼夜节律相关的基因（CCND1、FOXO1、FRMD8、NTRK2、PRRT1和TFPI）与IVDD进展及免疫相关。这些差异表达基因的识别可能为IVDD的诊断和治疗提供新的见解。
