JOR Spine最新文献_第10页

Macrophage Changes and High-Throughput Sequencing in Aging Mouse Intervertebral Disks 老龄小鼠椎间盘巨噬细胞变化和高通量测序

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-04-08 DOI: 10.1002/jsp2.70061

Wei Wang, Cheng Jiang, Jiong-Hui Chen, Yong-Long Chen, Zhen-Wu Zhang, Zhi-Chao Yang, Jun Li, Xiao-Chuan Li

Background

Intervertebral disk (IVD) degeneration is associated with lower back pain and aging; however, the mechanisms underlying age-related changes and the changes in macrophage polarization in aging intervertebral disks require further elucidation. The aim of this study was to evaluate changes in macrophages, the differential expression of senescence genes, and their relationship with hub genes in IVDs during aging in mice.

Methods

Twenty-eight male wild C57 mice aged 4 weeks were divided into two groups. Four mice per group were selected for high-throughput sequencing and 10 for tail IVD immunohistochemical analysis. Adult and aged mouse IVD specimens were stained with hematoxylin–eosin, Fast Green, and Alcian Blue to determine collagen (Col) 1, Col2, proteoglycan, P16, P21, P53, CD11b, CD86, CD206, IL-1, TGF-β, and IL-4 expression. High-throughput sequencing was performed on adult and aged mouse IVD tissues.

Results

Aged mouse IVDs showed reduced height and marked degeneration, with decreased Col2 and proteoglycan expression and increased Col1 expression. The expression of senescence markers, senescence-associated IL-1, TGF-β, and IL-4, and macrophage-related markers, CD11b, CD86, and CD206, increased markedly with age. High-throughput sequencing revealed 1975 differentially expressed genes in adult and aged mice, with 797 genes showing upregulated expression (top five: Kcna7, Mmp9, Panx3, Myl10, and Bglap) and 1178 showing downregulated expression (top five: Srd5a2, Slc38a5, Gm47283, Npy, and Pcdh8). Gene Ontology and pathway enrichment analyses highlighted aging-related cellular components, biological processes, and metabolic pathways. The identified hub genes included Cox5a, Ndufs6, and Ndufb9.

Conclusions

Disk senescence and reduced height in aged mice are linked to upregulated expression of senescence-associated phenotypes and macrophage polarization markers. These findings suggest that macrophages and differential gene expression play key roles in age-related IVD degeneration, indicating that they can be used as potential targets for therapeutic intervention.

椎间盘（IVD）退变与下背部疼痛和衰老有关；然而，衰老椎间盘中年龄相关变化和巨噬细胞极化变化的机制需要进一步阐明。本研究旨在探讨小鼠ivd衰老过程中巨噬细胞的变化、衰老基因的差异表达及其与中枢基因的关系。方法4周龄雄性野生C57小鼠28只，随机分为2组。每组选取4只小鼠进行高通量测序，10只小鼠进行尾IVD免疫组化分析。成年和老年小鼠IVD标本采用苏木精-伊红、Fast Green和Alcian Blue染色，检测胶原（Col） 1、Col2、蛋白聚糖、P16、P21、P53、CD11b、CD86、CD206、IL-1、TGF-β和IL-4的表达。对成年和老年小鼠IVD组织进行高通量测序。结果老龄小鼠ivd高度降低，变性明显，Col2和蛋白多糖表达降低，Col1表达升高。衰老标志物衰老相关IL-1、TGF-β、IL-4和巨噬细胞相关标志物CD11b、CD86、CD206的表达随着年龄的增长而显著升高。高通量测序结果显示，在成年和老年小鼠中，有1975个差异表达基因，其中797个基因表达上调（前5个：Kcna7、Mmp9、Panx3、Myl10和Bglap）， 1178个基因表达下调（前5个：Srd5a2、Slc38a5、Gm47283、Npy和Pcdh8）。基因本体和途径富集分析强调了与衰老相关的细胞成分、生物过程和代谢途径。中心基因包括Cox5a、Ndufs6和Ndufb9。结论衰老小鼠的椎间盘衰老和身高下降与衰老相关表型和巨噬细胞极化标记物的表达上调有关。这些发现表明巨噬细胞和差异基因表达在年龄相关的IVD变性中起关键作用，表明它们可以作为治疗干预的潜在靶点。

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引用次数: 0

Evidence for a Causal Association Between Human Cytomegalovirus Infection and Chronic Back Pain: A One-Sample Mendelian Randomization Study 人类巨细胞病毒感染与慢性背痛之间因果关系的证据：一项单样本孟德尔随机研究

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-04-08 DOI: 10.1002/jsp2.70063

Maryam Kazemi Naeini, Maxim B. Freidin, Isabelle Granville Smith, Stephen Ward, Frances M. K. Williams

<div> <section> <h3> Background</h3> <p>Chronic back pain (CBP) is a major cause of disability globally. While its etiology is multifactorial, specific contributing genetic and environmental factors remain to be discovered. Paraspinal muscle fat has been shown in human and preclinical studies to be related to CBP. One potential risk factor is infection by cytomegalovirus (CMV) because CMV is trophic for fat. CMV may reside in the paraspinal muscle adipose tissue. We set out to test the hypothesis that previous CMV infection is linked to CPB using a one-sample Mendelian randomization (MR).</p> </section> <section> <h3> Method</h3> <p>The sample comprised 5140 UK Biobank participants with information about CMV serology and CBP status. A one-sample MR based on independent genetic variants predicting CMV positivity was conducted in Northern European participants. To validate the association further, the MR study was repeated using a CMV polygenic risk score (PRS). As a negative control for confounding and spurious causal inference, we used Epstein–Barr virus (EBV) serology, because EBV is another common viral infection but is not trophic for adipose tissue.</p> </section> <section> <h3> Results</h3> <p>A genome-wide association study for CMV seropositivity revealed 86 independent SNPs having p-value < <span></span><math> <semantics> <mrow> <mn>2</mn> <mo>×</mo> <msup> <mn>10</mn> <mrow> <mo>−</mo> <mn>4</mn> </mrow> </msup> </mrow> <annotation>$$ 2times {10}^{-4} $$</annotation> </semantics></math> that have been used to define genetically-predicted categories of CMV infection risk. The CMV predicted categories were found statistically significantly associated with CBP (OR = 1.150; 95% CI: 1.005–1.317, <i>p</i>-value = 0.043). Stronger significant results were obtained using the PRS for CMV seropositivity (OR = 1.290; 95% CI: 1.133–1.469, <i>p</i>-value = 12E-4). No such association was seen between EBV and CBP.</p> </section> <section> <h3> Conclusion</h3> <p>Our results provide evidence for a causal relationship between CMV infection and CBP. Further investigation is warranted to get insight into the mechanism by which CMV might contribute to the pathogenesis of CBP.</p>

慢性背痛（CBP）是全球致残的主要原因之一。虽然其病因是多因素的，但具体的遗传和环境因素仍有待发现。在人体和临床前研究中，棘旁肌脂肪已被证明与CBP有关。一个潜在的危险因素是感染巨细胞病毒（CMV），因为巨细胞病毒对脂肪有营养作用。巨细胞病毒可能存在于棘旁肌肉脂肪组织中。我们开始使用单样本孟德尔随机化（MR）来验证先前的巨细胞病毒感染与CPB相关的假设。方法选取5140名英国生物样本库参与者，收集CMV血清学和CBP状态信息。在北欧参与者中进行了基于独立遗传变异预测巨细胞病毒阳性的单样本MR。为了进一步验证相关性，使用CMV多基因风险评分（PRS）重复MR研究。作为混淆和虚假因果推断的阴性对照，我们使用eb病毒（EBV）血清学，因为EBV是另一种常见的病毒感染，但对脂肪组织没有营养。结果CMV血清阳性的全基因组关联研究显示，86个独立snp的p值为＆lt； 2 × 10−4$$ 2times {10}^{-4} $$已被用于定义CMV感染风险的遗传预测类别。CMV预测类别与CBP有统计学显著相关(OR = 1.150；95% CI: 1.005–1.317, p-value = 0.043). Stronger significant results were obtained using the PRS for CMV seropositivity (OR = 1.290; 95% CI: 1.133–1.469, p-value = 12E-4). No such association was seen between EBV and CBP. Conclusion Our results provide evidence for a causal relationship between CMV infection and CBP. Further investigation is warranted to get insight into the mechanism by which CMV might contribute to the pathogenesis of CBP.

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引用次数: 0

Molecular Subtypes and Immune Microenvironment Characterization of the Annulus Fibrosus in Intervertebral Disc Degeneration: Insights From Translation Factor-Related Gene Analysis 椎间盘退变中纤维环分子亚型和免疫微环境特征：来自翻译因子相关基因分析的见解

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-04-07 DOI: 10.1002/jsp2.70064

Sikuan Zheng, Xiaokun Zhao, Hui Wu, Xuhui Cuan, Xigao Cheng, Dingwen He

Objective

This study aims to examine the role of translation factors (TF) in intervertebral disc degeneration (IVDD) and to evaluate their clinical relevance through unsupervised clustering methods.

Methods

Gene expression data were retrieved from the GEO database, and the expression levels of translation factor-related genes (TFGs) were extracted for analysis.

Results

Two distinct molecular clusters were identified based on the differential expression of nine significantly altered TFGs. Immune infiltration was notably higher in Cluster C2 compared to Cluster C1. Subsequently, two gene clusters were identified based on the differentially expressed genes between the clusters. A Sankey diagram illustrated a high degree of consistency between the molecular clusters and the gene clusters. Additionally, four machine learning models were developed and evaluated, with the SVM model being utilized to construct a nomogram for predicting the incidence of IVDD. Validation using external datasets and clinical samples confirmed the low expression of EEF2K, which was further analyzed in a pan-cancer context.

Conclusion

The identification and comprehensive assessment of the two molecular clusters offer significant insights for the classification and treatment of individuals with IVDD.

目的探讨翻译因子（TF）在椎间盘退变（IVDD）中的作用，并通过无监督聚类方法评价其临床相关性。方法从GEO数据库中检索基因表达数据，提取翻译因子相关基因（TFGs）的表达水平进行分析。结果基于9个显著改变的TFGs的差异表达，鉴定出两个不同的分子簇。C2组的免疫浸润明显高于C1组。随后，根据两个基因簇之间的差异表达基因，鉴定出两个基因簇。桑基图说明了分子簇和基因簇之间的高度一致性。此外，开发并评估了四种机器学习模型，并利用SVM模型构建了预测IVDD发生率的nomogram。使用外部数据集和临床样本验证证实EEF2K的低表达，并在泛癌症背景下进一步分析。结论两个分子簇的鉴别和综合评价对IVDD患者的分类和治疗具有重要意义。

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引用次数: 0

Analysis of Nicotine Toxicity and Mechanisms of Senescence in Nucleus Pulposus Cells Using Network Toxicology and Molecular Docking Technique 利用网络毒理学和分子对接技术分析烟碱对髓核细胞的毒性及衰老机制

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-31 DOI: 10.1002/jsp2.70055

Chen Jiang, Chao Song, Chaoqi Chen, Baoxin Shen, Lei Yang, Chi Zhang, Fei Liu, Xiaofei Wu, Feng Chen

Aim

Through the use of network toxicology, the research sought to determine whether cellular senescence and associated molecular mechanisms in nicotine-induced intervertebral disc degeneration (IVDD) were potentially harmful.

Methods

The primary chemical structure and 105 targets of action of nicotine were determined by using the Swiss Target Prediction, Cell Age, and PubChem databases. 855 IVDD senescence genes were found using the GEO and Cell Age datasets.

Results

After additional screening and Cytoscape development, 9 key targets were identified. Additionally, these targets' co-expression pattern analysis and protein interactions were confirmed to be identical. The core targets of nicotine-induced IVDD cellular senescence were found to be primarily enriched in the positive regulation of cell proliferation, telomere shortening, histone acetylation, and cellular senescence-related processes, according to gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). The KEGG signaling pathway also made it clear that the Apelin signaling route, nicotinate and nicotinamide metabolism, cell cycle, and apoptosis are all strongly linked to nicotine-induced IVDD cellular senescence. We chose four genes associated with the cellular senescence pathway—HDAC1, HDAC4, and NAMPT, MYLK—for molecular docking with the toxic substance nicotine. The findings validated nicotine's strong affinity for the primary targets.

Conclusion

All things considered, the current research indicates that nicotine may contribute to cellular senescence in IVDD via controlling the histone deacetylation process, telomere shortening, the Apelin signaling pathway, and pathways linked to the metabolism of nicotinate and nicotinamide. The theoretical foundation for investigating the molecular mechanisms of nicotine-induced senescence in IVDD is established.

目的通过网络毒理学的应用，本研究试图确定尼古丁诱导的椎间盘退变（IVDD）的细胞衰老及其相关分子机制是否具有潜在的危害性。方法利用Swiss Target Prediction、Cell Age和PubChem数据库对尼古丁的主要化学结构和105个作用靶点进行测定。使用GEO和Cell Age数据集发现了855个IVDD衰老基因。结果经进一步筛选和细胞景观开发，鉴定出9个关键靶点。此外，这些靶点的共表达模式分析和蛋白质相互作用被证实是相同的。根据基因本体（GO）和京都基因与基因组百科全书（KEGG），尼古丁诱导IVDD细胞衰老的核心靶点主要富集于细胞增殖、端粒缩短、组蛋白乙酰化和细胞衰老相关过程的正调控。KEGG信号通路也明确了Apelin信号通路、烟酸和烟酰胺代谢、细胞周期和细胞凋亡都与尼古丁诱导的IVDD细胞衰老密切相关。我们选择了四个与细胞衰老途径相关的基因——hdac1、HDAC4和NAMPT、mylk——与有毒物质尼古丁进行分子对接。这些发现证实了尼古丁对主要目标的强烈亲和力。综上所述，目前的研究表明，尼古丁可能通过控制组蛋白去乙酰化过程、端粒缩短、Apelin信号通路以及与烟酸和烟酰胺代谢相关的通路，促进IVDD的细胞衰老。为研究尼古丁诱导IVDD衰老的分子机制奠定了理论基础。

{"title":"Analysis of Nicotine Toxicity and Mechanisms of Senescence in Nucleus Pulposus Cells Using Network Toxicology and Molecular Docking Technique","authors":"Chen Jiang, Chao Song, Chaoqi Chen, Baoxin Shen, Lei Yang, Chi Zhang, Fei Liu, Xiaofei Wu, Feng Chen","doi":"10.1002/jsp2.70055","DOIUrl":"https://doi.org/10.1002/jsp2.70055","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>Through the use of network toxicology, the research sought to determine whether cellular senescence and associated molecular mechanisms in nicotine-induced intervertebral disc degeneration (IVDD) were potentially harmful.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The primary chemical structure and 105 targets of action of nicotine were determined by using the Swiss Target Prediction, Cell Age, and PubChem databases. 855 IVDD senescence genes were found using the GEO and Cell Age datasets.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>After additional screening and Cytoscape development, 9 key targets were identified. Additionally, these targets' co-expression pattern analysis and protein interactions were confirmed to be identical. The core targets of nicotine-induced IVDD cellular senescence were found to be primarily enriched in the positive regulation of cell proliferation, telomere shortening, histone acetylation, and cellular senescence-related processes, according to gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). The KEGG signaling pathway also made it clear that the Apelin signaling route, nicotinate and nicotinamide metabolism, cell cycle, and apoptosis are all strongly linked to nicotine-induced IVDD cellular senescence. We chose four genes associated with the cellular senescence pathway—HDAC1, HDAC4, and NAMPT, MYLK—for molecular docking with the toxic substance nicotine. The findings validated nicotine's strong affinity for the primary targets.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>All things considered, the current research indicates that nicotine may contribute to cellular senescence in IVDD via controlling the histone deacetylation process, telomere shortening, the Apelin signaling pathway, and pathways linked to the metabolism of nicotinate and nicotinamide. The theoretical foundation for investigating the molecular mechanisms of nicotine-induced senescence in IVDD is established.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 2","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70055","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143741645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Phosphatidylcholine Ameliorates Palmitic Acid-Induced Lipotoxicity by Facilitating Endoplasmic Reticulum and Mitochondria Contacts in Intervertebral Disc Degeneration 磷脂酰胆碱通过促进椎间盘退变中内质网和线粒体接触改善棕榈酸诱导的脂肪毒性

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-31 DOI: 10.1002/jsp2.70062

Shuangshuang Tu, Yijun Dong, Chuanfu Li, Mingxin Jiang, Liqun Duan, Wenzhi Zhang, Xi Chen

Background

Intervertebral disc degeneration (IDD) is a prevalent musculoskeletal disorder with substantial socioeconomic impacts. Despite its high prevalence, the pathogenesis of IDD remains unclear, and effective pharmacological interventions are lacking. This study aimed to investigate metabolic alterations in IDD and explore potential therapeutic targets by analyzing lipotoxicity-related mechanisms in nucleus pulposus (NP) cells.

Methods

Metabolomics and magnetic resonance spectroscopy were utilized to profile metabolic changes in NP tissues from advanced-stage IDD. Transcriptomics and metabolomics integration were performed to identify key regulatory pathways. In vitro experiments using human NP cells exposed to palmitic acid were conducted to evaluate endoplasmic reticulum (ER) stress, mitochondrial dysfunction, lipid droplet accumulation, and senescence. Phosphatidylcholine supplementation was tested for its ability to mitigate lipotoxicity, with ER-mitochondria interactions and mitochondrial oxidation capacity assessed as mechanistic endpoints.

Results

Our findings revealed an abnormal lipotoxic condition in NP cells from advanced-stage IDD. Furthermore, we identified abnormal accumulation of triglycerides and palmitic acid in NP cells from IDD. The palmitic acid accumulation resulted in endoplasmic reticulum stress, mitochondrial damage, lipid droplet accumulation, and senescence of NP cells. By integrating transcriptomics and metabolomics analyses, we discovered that phosphatidylcholine plays a role in regulating palmitic acid-induced lipotoxicity. Notably, phosphatidylcholine level was found to be low in the endoplasmic reticulum and mitochondria of advanced-stage NP cells. Phosphatidylcholine treatment alleviated palmitic acid-induced lipid droplet accumulation and senescence of NP cells by modulating ER-mitochondria contacts and mitochondrial oxidation capacity.

Conclusion

Phosphatidylcholine emerges as a potential therapeutic agent to counteract lipotoxic stress by modulating organelle interactions and mitochondrial function. These findings advance our understanding of IDD pathogenesis and provide a novel metabolic target for therapeutic development.

背景椎间盘退变（IDD）是一种普遍存在的肌肉骨骼疾病，具有重大的社会经济影响。尽管发病率很高，但IDD的发病机制尚不清楚，缺乏有效的药物干预措施。本研究旨在通过分析髓核（NP）细胞脂毒相关机制，研究IDD的代谢改变，并探索潜在的治疗靶点。方法利用代谢组学和磁共振波谱分析晚期IDD患者NP组织的代谢变化。转录组学和代谢组学整合来确定关键的调控途径。我们利用暴露于棕榈酸的人NP细胞进行了体外实验，以评估内质网（ER）应激、线粒体功能障碍、脂滴积累和衰老。磷脂酰胆碱补充剂被测试其减轻脂肪毒性的能力，并以内质网线粒体相互作用和线粒体氧化能力作为机制终点进行评估。结果发现晚期IDD患者NP细胞存在异常脂毒性。此外，我们在IDD的NP细胞中发现了甘油三酯和棕榈酸的异常积累。棕榈酸积累导致NP细胞内质网应激、线粒体损伤、脂滴积累和衰老。通过整合转录组学和代谢组学分析，我们发现磷脂酰胆碱在调节棕榈酸诱导的脂肪毒性中起作用。值得注意的是，在晚期NP细胞的内质网和线粒体中发现磷脂酰胆碱水平低。磷脂酰胆碱处理通过调节er -线粒体接触和线粒体氧化能力减轻棕榈酸诱导的NP细胞脂滴积累和衰老。结论磷脂酰胆碱可能通过调节细胞器相互作用和线粒体功能来对抗脂毒性应激。这些发现促进了我们对IDD发病机制的理解，并为治疗开发提供了新的代谢靶点。

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引用次数: 0

Transcriptome Data Combined With Mendelian Randomization Analysis Identifies Key Genes Associated With Mitochondria and Programmed Cell Death in Intervertebral Disc Degeneration 转录组数据结合孟德尔随机化分析确定与椎间盘退变中线粒体和程序性细胞死亡相关的关键基因

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-24 DOI: 10.1002/jsp2.70057

Hongfei Nie, Xiao Hu, Jiaxiao Wang, Jia Wang, Xiaoqian Yu, Jun Li

Background

Intervertebral disc degeneration (IDD) is a major cause of cervical and lumbar diseases, significantly impacting patients' quality of life. Mitochondria and cell death have been implicated in IDD, but the key related genes remain unknown.

Methods

Differentially expressed genes (DEGs) between IDD and control samples were identified using GSE70362. Mitochondria-related genes (MRGs) and programmed cell death-related genes (PCDRGs) were intersected with DEGs to find DE-MRGs and DE-PCDRGs. Weighted gene co-expression network analysis (WGCNA) identified key module genes, and the overlap with DEGs revealed candidate genes. Mendelian randomization (MR) analysis was used to determine genes causally linked to IDD. Machine learning and expression validation further refined key genes, which were then used to build a nomogram to predict IDD risk. Additionally, gene set enrichment analysis (GSEA), immune infiltration, and single-cell analysis were performed.

Results

A total of 515 DEGs were intersected with 224 key module genes, yielding 31 candidate genes. Six genes—BCKDHB, BID, TNFAIP6, VRK1, CAB39L, and TMTC1—showed a causal relationship with IDD. BID, TNFAIP6, and TMTC1 were further identified as key genes through machine learning and validation. A nomogram was developed based on these genes. GSEA revealed BID and TMTC1 were enriched in N-glycan biosynthesis, TNFAIP6 and TMTC1 in aminoacyl tRNA biosynthesis, and BID and TMTC1 in ribosomal pathways. Activated dendritic cells, CD56dim natural killer cells, monocytes, and other immune cells were elevated in IDD, with TNFAIP6 strongly correlating with activated dendritic cells. Key genes were expressed at higher levels in degraded samples.

Conclusion

BID, TMTC1, and TNFAIP6 were identified as key genes linked to mitochondria and cell death in IDD, offering new insights for diagnosis and treatment.

背景椎间盘退变（IDD）是导致颈腰椎疾病的主要原因，严重影响患者的生活质量。线粒体和细胞死亡与缺乏症有关，但关键的相关基因尚不清楚。方法用GSE70362检测IDD与对照的差异表达基因（DEGs）。线粒体相关基因（MRGs）和程序性细胞死亡相关基因（PCDRGs）与deg相交，发现DE-MRGs和DE-PCDRGs。加权基因共表达网络分析（Weighted gene co-expression network analysis， WGCNA）确定了关键模块基因，与deg的重叠揭示了候选基因。采用孟德尔随机化（MR）分析确定与缺乏症相关的基因。机器学习和表达验证进一步细化了关键基因，然后用这些基因构建一个nomogram来预测IDD风险。此外，还进行了基因集富集分析（GSEA）、免疫浸润和单细胞分析。结果共获得515个deg与224个关键模块基因相交，得到31个候选基因。bckdhb、BID、TNFAIP6、VRK1、CAB39L、tmtc1等6个基因与IDD存在因果关系。通过机器学习和验证进一步确定BID、TNFAIP6和TMTC1为关键基因。在这些基因的基础上形成了一种形态图。GSEA显示BID和TMTC1在n -聚糖生物合成中富集，TNFAIP6和TMTC1在氨基tRNA生物合成中富集，BID和TMTC1在核糖体途径中富集。激活的树突状细胞、CD56dim自然杀伤细胞、单核细胞和其他免疫细胞在IDD中升高，TNFAIP6与活化的树突状细胞密切相关。关键基因在降解样品中表达水平较高。结论BID、TMTC1和TNFAIP6是IDD中线粒体和细胞死亡相关的关键基因，为IDD的诊断和治疗提供了新的思路。

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引用次数: 0

The Significance of MAPK Signaling Pathway in the Diagnosis and Subtype Classification of Intervertebral Disc Degeneration MAPK信号通路在椎间盘退变诊断及亚型分型中的意义

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-24 DOI: 10.1002/jsp2.70060

Yong Liu, Xueyan Chen, Jingwen Chen, Chao Song, Zhangchao Wei, Zongchao Liu, Fei Liu

Background

Intervertebral disc degeneration (IDD) is a human aging disease related mainly to inflammation, cellular senescence, RNA/DNA methylation, and ECM. The mitogen-activated protein kinase (MAPK) signaling pathway is engaged in multiple biological functions by phosphorylating specific serine and threonine residues on target proteins through phosphorylation cascade effects, but the role and specific mechanisms of the MAPK signaling pathway in IDD are still unclear.

Methods

We identified 20 MAPK-related differential genes by differential analysis of the GSE124272 and GSE150408 datasets from the GEO database. To explore the biological functions of these differential genes in humans, we performed GO and KEGG analyses. Additionally, we applied PPI networks, LASSO analysis, the RF algorithm, and the SVM-RFE algorithm to identify core MAPK-related genes. Finally, we conducted further validation using clinical samples.

Results

We ultimately identified and validated four pivotal MAPK-related genes, namely, KRAS, JUN, RAP1B, and TNF, using clinical samples, and constructed the ROC curves to evaluate the predictive accuracy of the hub genes. A nomogram model was subsequently developed based on these four hub MAPK genes to predict the prevalence of IDD. Based on these four hub genes, we classified IDD patients into two MAP clusters by applying the consensus clustering method and identified 1916 DEGs by analyzing the differences between the two clusters. Further analysis using the same approach allowed us to identify two gene clusters based on these DEGs. We used a PCA algorithm to determine the MAPK score for each sample and discovered that MAPK cluster A and gene cluster A had higher scores, suggesting greater sensitivity to MAPK signaling pathway-associated agents in the subtype. We displayed the differing expression levels of four hub MAPK-related genes across the two clusters and their relationship with immune cell infiltration to highlight the distinctions between clusters A and B.

Conclusion

In summary, four hub MAPK signaling pathway-related genes, KRAS, JUN, RAP1B, and TNF, could be applied to the diagnosis and subtype classification of IDD and benefit the prevention and treatment of IDD.

椎间盘退变（IDD）是一种人类老年性疾病，主要与炎症、细胞衰老、RNA/DNA甲基化和ECM有关。丝裂原活化蛋白激酶（MAPK）信号通路通过磷酸化级联效应磷酸化靶蛋白上的特定丝氨酸和苏氨酸残基，参与多种生物学功能，但MAPK信号通路在IDD中的作用和具体机制尚不清楚。方法通过对GEO数据库中GSE124272和GSE150408数据集的差异分析，鉴定出20个mapk相关差异基因。为了探索这些差异基因在人类中的生物学功能，我们进行了GO和KEGG分析。此外，我们应用PPI网络、LASSO分析、RF算法和SVM-RFE算法来识别核心mapk相关基因。最后，我们使用临床样本进行了进一步的验证。结果我们最终通过临床样本鉴定并验证了KRAS、JUN、RAP1B和TNF四个关键mapk相关基因，并构建了ROC曲线来评估枢纽基因的预测准确性。随后，基于这四个中心MAPK基因建立了一个nomogram模型来预测IDD的患病率。基于这4个中心基因，采用共识聚类法将IDD患者分为两个MAP聚类，并通过分析两个聚类之间的差异，鉴定出1916个deg。使用相同方法的进一步分析使我们能够根据这些deg确定两个基因簇。我们使用PCA算法确定每个样本的MAPK得分，发现MAPK集群a和基因集群a得分更高，表明该亚型对MAPK信号通路相关药物更敏感。我们展示了4个枢纽MAPK相关基因在两个集群中的不同表达水平及其与免疫细胞浸润的关系，以突出集群A和b之间的差异。结论综上所述，4个枢纽MAPK信号通路相关基因KRAS、JUN、RAP1B和TNF。可用于IDD的诊断和亚型分类，有利于IDD的预防和治疗。

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引用次数: 0

Application and Validation of Semiautomatic Quantification of Immunohistochemically Stained Sections for Low Cellular Tissue Such as Intervertebral Disc Using QuPath QuPath在低细胞组织（如椎间盘）免疫组织化学染色切片半自动定量分析中的应用与验证

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-06 DOI: 10.1002/jsp2.70054

Andrea Nüesch, Maria Paola Ferri, Christine L. Le Maitre

Background

Immunohistochemistry (IHC) is a widely used method for localizing and semi-quantifying proteins in tissue samples. Traditional IHC analysis often relies on manually counting 200 cells within a designated area, a time-intensive and subjective process that can compromise reproducibility and accuracy. Advances in digital scanning and bioimage analysis tools, such as the open-source software QuPath, enable semi-automated cell counting, reducing subjectivity and increasing efficiency.

Aims

This project developed a QuPath-based script and detailed guide for semi-automatic cell counting, specifically for tissues with low cellularity, such as intervertebral discs and cartilage.

Methods and Results

The methodology was validated by demonstrating no significant differences between the manual counting and the semi-automatic quantification (p = 0.783, p = 0.386) while showing a strong correlation between methods for both collagen type II staining (r = 0.9602, p < 0.0001) and N-cadherin staining (r = 0.9044, p = 0.0001). Furthermore, a strong correlation (intraclass correlation coefficient (ICC) single raters = 0.853) between 3 individual raters with varying academic ranks and experiences in IHC analysis was shown using the semi-automatic quantification method.

Discusssion

The approach ensures high reproducibility and accuracy, with reduced variability between raters and laboratories. This semi-automated method is particularly suited for tissues with a high extracellular matrix to cell ratio and low cellularity. By minimizing subjectivity and evaluation time, it provides a robust alternative to manual counting, making it ideal for applications where reproducibility and standardization are critical. While the methodology was effective in low-cellularity tissues, its application in other tissue types warrants further exploration.

Conclusions

These findings underscore the potential of QuPath to streamline IHC analysis and enhance inter-laboratory comparability.

免疫组织化学（IHC）是一种广泛应用于组织样品中蛋白质定位和半定量的方法。传统的免疫组化分析通常依赖于在指定区域内手动计数200个细胞，这是一个耗时且主观的过程，可能会损害再现性和准确性。数字扫描和生物图像分析工具的进步，如开源软件QuPath，实现了半自动化细胞计数，减少了主观性，提高了效率。本项目开发了一个基于qupath的脚本和详细指南，用于半自动细胞计数，特别是对于低细胞组织，如椎间盘和软骨。方法与结果手工计数与半自动定量之间无显著差异（p = 0.783, p = 0.386），而II型胶原染色方法（r = 0.9602, p < 0.0001）与N-cadherin染色方法（r = 0.9044, p = 0.0001）之间有很强的相关性。此外，采用半自动化的定量方法，3个不同学术等级和免疫结构分析经验的评分者之间具有很强的相关性（类内相关系数(ICC) = 0.853）。该方法确保了高再现性和准确性，减少了评分者和实验室之间的可变性。这种半自动化的方法特别适合于具有高细胞外基质与细胞比例和低细胞密度的组织。通过最大限度地减少主观性和评估时间，它为手动计数提供了一个健壮的替代方案，使其成为对再现性和标准化至关重要的应用程序的理想选择。虽然该方法在低细胞组织中有效，但其在其他组织类型中的应用仍有待进一步探索。这些发现强调了QuPath在简化IHC分析和增强实验室间可比性方面的潜力。

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引用次数: 0

The Significance of Cellular Senescence Hub Genes in the Diagnosis and Subtype Classification of a Comprehensive Database of Gene Expression in Intervertebral Disc Degeneration 细胞衰老中枢基因在椎间盘退变诊断和基因表达综合数据库亚型分类中的意义

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-06 DOI: 10.1002/jsp2.70050

Fei Liu, Silong Gao, Ji Yin, Chao Song, Yongliang Mei, Zhaoqiang Wang, Zongchao Liu

Background

Intervertebral disc degeneration (IVDD) is a complex age-related physiological process, with cellular senescence (CS) being a primary contributing factor. However, the precise role of CS and its associated genes in IVDD remains unclear.

Methods

In this study, we performed differential expression analysis on the GSE124272 and GSE150408 datasets from the GEO database and identified 53 differentially expressed cellular senescence-related genes (CSRGs). We then conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to explore their functions and associated pathways. We identified hub genes by constructing a protein–protein interaction (PPI) network and further validated these genes using clinical samples. We further explored the functional and prognostic significance of these genes using support vector machine recursive feature elimination (SVM-RFE), random forest (RF), and least absolute shrinkage and selection operator (LASSO) algorithms. We visualized the correlation between the differential expression levels of the four core genes and immune cell infiltration using heat maps and histograms. Finally, we performed graphene oxide enrichment analysis on 297 differentially expressed genes (DEGs) to investigate their role in IVDD.

Results

We ultimately identified four hub cellular CSRGs DUSP3, MAPKAPK5, SP1, and VEGFA, and further validated their expression using various algorithms and clinical samples. Our results revealed that DUSP3 and SP1 were upregulated in IVDD, while MAPKAPK5 and VEGFA were downregulated. Immune cell infiltration analysis demonstrated that DUSP3 and SP1 were positively correlated with immune cell infiltration levels, whereas VEGFA and MAPKAPK5 were negatively correlated.

Conclusion

In summary, CSRGs play an important role in the pathogenesis of IVDD, and our study of the hub gene cluster may guide future therapeutic strategies for IVDD.

椎间盘退变（IVDD）是一个复杂的与年龄相关的生理过程，细胞衰老（CS）是一个主要因素。然而，CS及其相关基因在IVDD中的确切作用尚不清楚。方法对GEO数据库中的GSE124272和GSE150408数据集进行差异表达分析，鉴定出53个差异表达的细胞衰老相关基因（CSRGs）。然后，我们进行了基因本体（GO）和京都基因与基因组百科全书（KEGG）途径富集分析，以探索它们的功能和相关途径。我们通过构建蛋白-蛋白相互作用（PPI）网络来确定枢纽基因，并通过临床样本进一步验证这些基因。我们使用支持向量机递归特征消除（SVM-RFE）、随机森林（RF）和最小绝对收缩和选择算子（LASSO）算法进一步探讨了这些基因的功能和预后意义。我们使用热图和直方图可视化了四个核心基因的差异表达水平与免疫细胞浸润之间的相关性。最后，我们对297个差异表达基因（DEGs）进行了氧化石墨烯富集分析，以研究它们在IVDD中的作用。我们最终鉴定出4个中枢细胞CSRGs DUSP3、MAPKAPK5、SP1和VEGFA，并通过各种算法和临床样本进一步验证了它们的表达。我们的研究结果显示，DUSP3和SP1在IVDD中上调，MAPKAPK5和VEGFA下调。免疫细胞浸润分析表明，DUSP3和SP1与免疫细胞浸润水平呈正相关，而VEGFA和MAPKAPK5呈负相关。综上所述，CSRGs在IVDD的发病机制中发挥了重要作用，我们对枢纽基因簇的研究可以指导未来IVDD的治疗策略。

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引用次数: 0

DeVa (Decay Variance): A Novel Score Calculated via Postprocessing the Changes in Signal Intensity of an Intervertebral Disc in a T2* Multi-Echo Magnetic Resonance Image Can Quantify Painful and Degenerate Lumbar Vertebral Discs DeVa（衰减方差）：通过后处理T2*多回声磁共振图像中椎间盘信号强度的变化计算出的一种新的评分，可以量化疼痛和退变的腰椎间盘

IF 3.4 3区医学 Q1 ORTHOPEDICS

JOR Spine

Pub Date : 2025-03-06 DOI: 10.1002/jsp2.70056

Stone Sima, Alisha Sial, Suhani Sharma, Dheera Ananthakrishnan, Jeff Kuan, Ashish Diwan

Introduction

Low back pain (LBP), a global disability leader, is often linked to intervertebral disc (IVD) degeneration. Traditional diagnostics like T2-weighted MRI provide qualitative but imprecise evaluations. A novel post-processing MRI technique, Decay Variance (DeVa), has shown promise in differentiating degenerate from healthy discs in animal studies. DeVa quantifies IVD degeneration by analyzing variations in signal intensities within each voxel in a T2* 2D FLASH multi-echo MRI sequence. This study aimed to validate DeVa clinically and explore its correlation with pain severity.

Methods

A cross-sectional study included 77 chronic LBP patients and 8 controls, who underwent T2-weighted and T2* 2D FLASH MRI. DeVa scores (worst and sum of all discs) were recorded, alongside traditional assessments like disc bulge, stenosis, high-intensity zones, and Pfirrmann grade. Pain severity was measured with a numerical rating scale. Statistical analyses included Pearson correlation, t-tests, and Gardner-Altman plots to evaluate relationships between DeVa scores, degeneration, and pain.

Results

DeVa scores correlated strongly with Pfirrmann grade (r = 0.692, p < 0.001) and were significantly higher in discs with bulge, stenosis, or high-intensity zones (p < 0.001). Moderate correlations were observed between worst DeVa scores (r = 0.296, p < 0.01), total DeVa scores (r = 0.323, p < 0.005) and pain severity. Patients with chronic LBP without severe degeneration (Pfirrmann ≤ 3 with no stenosis observable on standard MRI) had significantly higher worst (1.38 ± 0.26 vs. 1.10 ± 0.29, p < 0.005) and total (5.39 ± 0.75 vs. 4.65 ± 0.61, p < 0.0.1) DeVa scores compared to controls.

Discussion

DeVa offers a quantitative, noninvasive approach to assessing IVD degeneration, showing strong correlations with disc health and pain. It demonstrates enhanced sensitivity over traditional MRI, enabling the identification of pain-generating discs and informing personalized treatment strategies for chronic LBP. Further validation in larger populations is needed.

下腰痛（LBP）是全球残疾的领导者，通常与椎间盘（IVD）退变有关。传统的诊断方法，如t2加权MRI，提供定性但不精确的评估。一种新的后处理MRI技术，衰减方差（DeVa），在动物研究中显示出了区分退变椎间盘和健康椎间盘的希望。DeVa通过分析T2* 2D FLASH多回波MRI序列中每个体素内信号强度的变化来量化IVD变性。本研究旨在临床验证DeVa并探讨其与疼痛严重程度的相关性。方法采用横断面研究方法，对77例慢性腰痛患者和8例对照组进行T2加权和T2* 2D FLASH MRI检查。记录DeVa评分（最差和所有椎间盘的总和），以及传统的评估，如椎间盘突出、狭窄、高强度区和Pfirrmann分级。疼痛严重程度用数值评定量表测量。统计分析包括Pearson相关、t检验和Gardner-Altman图来评估DeVa评分、退变和疼痛之间的关系。结果DeVa评分与Pfirrmann分级密切相关（r = 0.692, p < 0.001），在椎间盘突出、狭窄或高强度区明显较高（p < 0.001）。最差DeVa评分（r = 0.296, p < 0.01）、总DeVa评分（r = 0.323, p < 0.005）与疼痛严重程度呈正相关。无严重退变的慢性下腰痛患者（Pfirrmann≤3，标准MRI未观察到狭窄）的最差评分（1.38±0.26比1.10±0.29,p < 0.005）和总评分（5.39±0.75比4.65±0.61,p < 0.0.1）明显高于对照组。DeVa提供了一种定量的、无创的方法来评估IVD退变，显示与椎间盘健康和疼痛有很强的相关性。与传统MRI相比，它的灵敏度更高，能够识别产生疼痛的椎间盘，并为慢性腰痛的个性化治疗策略提供信息。需要在更大的人群中进一步验证。

{"title":"DeVa (Decay Variance): A Novel Score Calculated via Postprocessing the Changes in Signal Intensity of an Intervertebral Disc in a T2* Multi-Echo Magnetic Resonance Image Can Quantify Painful and Degenerate Lumbar Vertebral Discs","authors":"Stone Sima, Alisha Sial, Suhani Sharma, Dheera Ananthakrishnan, Jeff Kuan, Ashish Diwan","doi":"10.1002/jsp2.70056","DOIUrl":"https://doi.org/10.1002/jsp2.70056","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>Low back pain (LBP), a global disability leader, is often linked to intervertebral disc (IVD) degeneration. Traditional diagnostics like T2-weighted MRI provide qualitative but imprecise evaluations. A novel post-processing MRI technique, Decay Variance (DeVa), has shown promise in differentiating degenerate from healthy discs in animal studies. DeVa quantifies IVD degeneration by analyzing variations in signal intensities within each voxel in a T2* 2D FLASH multi-echo MRI sequence. This study aimed to validate DeVa clinically and explore its correlation with pain severity.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>A cross-sectional study included 77 chronic LBP patients and 8 controls, who underwent T2-weighted and T2* 2D FLASH MRI. DeVa scores (worst and sum of all discs) were recorded, alongside traditional assessments like disc bulge, stenosis, high-intensity zones, and Pfirrmann grade. Pain severity was measured with a numerical rating scale. Statistical analyses included Pearson correlation, <i>t</i>-tests, and Gardner-Altman plots to evaluate relationships between DeVa scores, degeneration, and pain.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>DeVa scores correlated strongly with Pfirrmann grade (<i>r</i> = 0.692, <i>p</i> < 0.001) and were significantly higher in discs with bulge, stenosis, or high-intensity zones (<i>p</i> < 0.001). Moderate correlations were observed between worst DeVa scores (<i>r</i> = 0.296, <i>p</i> < 0.01), total DeVa scores (<i>r</i> = 0.323, <i>p</i> < 0.005) and pain severity. Patients with chronic LBP without severe degeneration (Pfirrmann ≤ 3 with no stenosis observable on standard MRI) had significantly higher worst (1.38 ± 0.26 vs. 1.10 ± 0.29, <i>p</i> < 0.005) and total (5.39 ± 0.75 vs. 4.65 ± 0.61, <i>p</i> < 0.0.1) DeVa scores compared to controls.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Discussion</h3>\u0000 \u0000 <p>DeVa offers a quantitative, noninvasive approach to assessing IVD degeneration, showing strong correlations with disc health and pain. It demonstrates enhanced sensitivity over traditional MRI, enabling the identification of pain-generating discs and informing personalized treatment strategies for chronic LBP. Further validation in larger populations is needed.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70056","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143565145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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