Xia B, Xing J, Ai Q, Li H, Xu M, Hou T
Department of Orthopedics, First Affiliated Hospital of Army Medical University, Chongqing 400038, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Jun 20;41(6):883-890. doi: 10.12122/j.issn.1673-4254.2021.06.11.
To identify new therapeutic targets for intervertebral disc degeneration (IDD) by analyzing gene variations in IDD.
We analyzed surgical samples of intervertebral disc from 4 patients with IDD and 3 patients with non-IDD using RNA sequencing (RNA-seq) technology to identify significant differentially expressed genes (DEGs) in IDD. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were utilized for gene enrichment studies to acquire the key genes and signal pathways during IDD progression. The differential expressions of the identified genes in IDD were validated in clinical samples with qRT-PCR.
The transcriptome profile revealed 512 significant DEGs, which were enriched in terms of keratinization, extracellular matrix (ECM) components, growth factor binding, and inflammatory chemotaxis in GO analysis. The top 10 terms of KEGG enrichment included amoebiasis, viral protein interaction with cytokine and cytokine receptor, ECM-receptor interaction, IL-17 signaling pathway, cytokine-cytokine receptor interaction, TNF signaling pathway, AGE-RAGE signaling pathway in diabetic complications, PI3K-Akt signaling pathway, chemokine signaling pathway and estrogen signaling pathway. Thirteen DEGs selected as the targets for qRT-PCR validation showed significant differential expressions in IDD ( < 0.001), and their expression trends were all consistent with the results of RNA-seq. Among these genes, 10 genes showed significant intergroup fold change (Log2FoldChange>1).
ECM, growth factors, collagen components, inflammatory chemokines and such signal pathways as TNF-α and PI3K-Akt all have important contributions to IDD progression and may thus serve as new therapeutic targets for treatment of IDD.
通过分析椎间盘退变(IDD)中的基因变异来确定IDD的新治疗靶点。
我们使用RNA测序(RNA-seq)技术分析了4例IDD患者和3例非IDD患者的椎间盘手术样本,以确定IDD中显著差异表达的基因(DEG)。利用基因本体论(GO)和京都基因与基因组百科全书(KEGG)数据库进行基因富集研究,以获取IDD进展过程中的关键基因和信号通路。通过qRT-PCR在临床样本中验证所鉴定基因在IDD中的差异表达。
转录组图谱显示有512个显著的DEG,在GO分析中,这些基因在角质化、细胞外基质(ECM)成分、生长因子结合和炎症趋化方面富集。KEGG富集的前10个条目包括阿米巴病、病毒蛋白与细胞因子及细胞因子受体的相互作用、ECM-受体相互作用、IL-17信号通路、细胞因子-细胞因子受体相互作用、TNF信号通路、糖尿病并发症中的AGE-RAGE信号通路、PI3K-Akt信号通路、趋化因子信号通路和雌激素信号通路。选择用于qRT-PCR验证的13个DEG在IDD中显示出显著差异表达(<0.001),其表达趋势均与RNA-seq结果一致。在这些基因中,10个基因显示出显著的组间倍数变化(Log2FoldChange>1)。
ECM、生长因子、胶原蛋白成分、炎症趋化因子以及TNF-α和PI3K-Akt等信号通路均对IDD进展有重要作用,因此可能成为治疗IDD的新治疗靶点。