Lu Chuanlong, Wang Heng, Qiao Maolin, Chang Runze, Chen Jinshan, Li Lizheng, Fan Keyi, Yan Sheng, Zhang Ruijing, Dong Honglin
Department of Vascular Surgery, The Second Hospital, Shanxi Medical University, Taiyuan, 030001, PR China.
Department of Nephrology, The Second Hospital of Shanxi Medical University, Taiyuan, China.
Sci Rep. 2025 Aug 7;15(1):28976. doi: 10.1038/s41598-025-14781-8.
Previous research has shown that the formation of Neutrophil Extracellular Traps (NETs), mediated by neutrophils, leads to an increase in inflammatory cell response and cellular tissue damage during Abdominal Aortic Aneurysm (AAA). Building on this foundation, our primary objective is to identify and validate potential hub genes involved in NETs-mediated intrinsic cellular damage and the recruitment process of immune cells. We performed an analysis on the infiltration of immune cells within the AAA transcriptome dataset, employing CIBERSORT and ssGSEA algorithms (GSE183464). Next, the gene expression patterns associated with NETs formation (GSE178883) were utilized to examine the physiological processes in peripheral blood neutrophils post-PMA (Phorbol 12-myristate 13-acetate) induction. Subsequently, utilizing bioinformatics and machine learning algorithms, candidate crucial genes were identified within NETs-related genes and transcriptome datasets (GSE179828, GSE145200, GSE161464, GSE57691, GSE232911 and GSE166676). Following this, receiver operating characteristic curves were created to evaluate the diagnostic significance of these essential genes. Furthermore, the correlation between pivotal genes and immune cells was then examined using CIBERSORT and ssGSEA algorithms. By using weighted gene co-expression network analysis and machine learning algorithms, we ultimately identified 9 hub genes (MMP9, CXCR4, CYBB, TNFAIP3, PIK3CD, LTB, CXCL13, SELL, STAT4). Finally, gene expression was verified using immunohistochemistry, immunofluorescence, and enzyme-linked immunosorbent assay methods. We have identified nine hub genes related to neutrophil-mediated intrinsic cellular damage and the recruitment of immune cells in AAA. These findings could offer clues to the cellular mechanisms mediated by neutrophils in the progression of AAA.
先前的研究表明,由中性粒细胞介导的中性粒细胞胞外陷阱(NETs)的形成会导致腹主动脉瘤(AAA)期间炎症细胞反应增加和细胞组织损伤。在此基础上,我们的主要目标是识别和验证参与NETs介导的固有细胞损伤和免疫细胞募集过程的潜在枢纽基因。我们使用CIBERSORT和ssGSEA算法(GSE183464)对AAA转录组数据集中的免疫细胞浸润进行了分析。接下来,利用与NETs形成相关的基因表达模式(GSE178883)来研究佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)诱导后外周血中性粒细胞中的生理过程。随后,利用生物信息学和机器学习算法,在NETs相关基因和转录组数据集(GSE179828、GSE145200、GSE161464、GSE57691、GSE232911和GSE166676)中识别出候选关键基因。在此之后,创建了受试者工作特征曲线以评估这些关键基因的诊断意义。此外,随后使用CIBERSORT和ssGSEA算法检查关键基因与免疫细胞之间的相关性。通过使用加权基因共表达网络分析和机器学习算法,我们最终确定了9个枢纽基因(基质金属蛋白酶9、趋化因子受体4、细胞色素b-245β链、肿瘤坏死因子α诱导蛋白3、磷脂酰肌醇-3-激酶δ、淋巴毒素β、CXC趋化因子配体13、淋巴细胞功能相关抗原1、信号转导和转录激活因子4)。最后,使用免疫组织化学、免疫荧光和酶联免疫吸附测定方法验证基因表达。我们已经确定了与中性粒细胞介导的AAA固有细胞损伤和免疫细胞募集相关的9个枢纽基因。这些发现可能为中性粒细胞在AAA进展中介导的细胞机制提供线索。
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