Identification and validation of hub genes related to neutrophil extracellular traps-mediated cell damage and immune recruitment during abdominal aortic aneurysm.

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.