Department of Rheumatology and Immunology, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
Clinical Institute of Inflammation and Immunology, Frontiers Science Center for Disease-Related Molecular Network, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
Arthritis Res Ther. 2023 Apr 22;25(1):66. doi: 10.1186/s13075-023-03043-5.
Neutrophils have a critical role in the pathogenesis of rheumatoid arthritis (RA) with immune system dysfunction. However, the molecular mechanisms of this process mediated by neutrophils still remain elusive. The purpose of the present study is to identify hub genes in neutrophils for diagnosis and treatment of RA utilizing publicly available datasets.
Gene expression profiles were downloaded from the Gene Expression Omnibus, and batch-corrected and normalized expression data were obtained using the ComBat package. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were used to conduct significantly functional analysis and crucial pathways. The resulting co-expression genes modules and hub genes were generated based on the weighted gene co-expression network analysis and visualization by Cytoscape. Flow cytometry was conducted to detect reactive oxygen species (ROS) levels in neutrophils.
Neutrophils underwent transcriptional changes in synovial fluid (SF) of RA patients, different from peripheral blood of healthy controls or patients with RA. Especially, glycolysis, HIF-1 signaling, NADH metabolism, and oxidative stress were affected. These hub genes were strongly linked with classical glycolysis-related genes (ENO1, GAPDH, and PKM) responsible for ROS production. The antioxidant enzyme glutathione peroxidase 3 (GPX3), a ROS scavenger, was first identified as a hub gene in RA neutrophils. Neutrophils from patients with autoinflammatory and autoimmune diseases had markedly enhanced ROS levels, most notably in RA SF.
This research recognized hub genes and explored the characteristics of neutrophils in RA. Our findings suggest that the novel hub gene GPX3 is involved in the neutrophil-driven oxidative stress-mediated pathogenesis of RA. It has the potency to be a target for neutrophil-directed RA therapy.
中性粒细胞在免疫功能障碍导致的类风湿关节炎（RA）发病机制中起着关键作用。然而，中性粒细胞介导的这一过程的分子机制仍不清楚。本研究旨在利用公共数据集确定中性粒细胞中与 RA 诊断和治疗相关的枢纽基因。
从基因表达综合数据库下载基因表达谱，使用 ComBat 包进行批次校正和标准化表达数据。使用基因本体论和京都基因与基因组百科全书富集分析进行显著功能分析和关键途径分析。根据加权基因共表达网络分析和 Cytoscape 可视化生成共表达基因模块和枢纽基因。通过流式细胞术检测中性粒细胞中活性氧（ROS）的水平。
RA 患者滑膜液中的中性粒细胞发生了转录变化，与健康对照者或 RA 患者的外周血不同。特别是糖酵解、HIF-1 信号、NADH 代谢和氧化应激受到影响。这些枢纽基因与负责 ROS 产生的经典糖酵解相关基因（ENO1、GAPDH 和 PKM）密切相关。抗氧化酶谷胱甘肽过氧化物酶 3（GPX3），一种 ROS 清除剂，首次被确定为 RA 中性粒细胞中的枢纽基因。自身炎症性和自身免疫性疾病患者的中性粒细胞 ROS 水平显著升高，尤其是在 RA 滑膜液中。
本研究鉴定了枢纽基因，并探讨了 RA 中性粒细胞的特征。我们的研究结果表明，新型枢纽基因 GPX3 参与了中性粒细胞驱动的氧化应激介导的 RA 发病机制。它有可能成为靶向中性粒细胞的 RA 治疗的靶点。
