Work

Unveiling the Temporal Dynamics and Molecular Regulation Profiles of Neutrophil Extracellular Traps Following Spinal Cord Injury

Year: 2025

Type: article

Abstract: Background: Spinal cord injury (SCI) initiates secondary inflammatory processes that exacerbate tissue damage, with neutrophil extracellular traps (NETs) playing a significant role in amplifying these cascades. This study aimed to explore the temporal dynamics and key regulatory genes of NET formation in SCI to identify therapeutic targets. Methods: We integrated two transcriptomic datasets from the GEO database to identify differentially expressed NETs-related genes (NRGs) in SCI. WGCNA identified SCI-related modules, while GSVA assessed NET formation dynamics. Publicly available single-cell RNA sequencing data from the GEO database determined cell-specific expression patterns of key NRGs. Findings were validated through immunofluorescence, Western blot, and qPCR in a mouse SCI model. Regulatory networks were constructed, and potential therapeutic compounds were predicted using DSigDB and molecular docking. Results: We identified seven key NRGs (Casp1, Ccl3, Fcgr2b, Itgam, Itgb2, Tlr2, Tlr4) in SCI. GSVA revealed peak NET score at day 1 post-injury, with attenuation at days 3 and 7. Single-cell transcriptome analysis demonstrated predominant expression of these key genes in neutrophils during the acute phase, most prominently at 1 day post-injury, which coincides with the most pronounced neutrophil infiltration. Immunofluorescence and Western blot analyses confirmed significantly elevated NET formation at 1 day post-SCI. qPCR verified the expression of all key NRGs. Regulatory network analysis identified CHD1 as an important transcription factor governing NET formation, while miRNA-mRNA network construction revealed sophisticated post-transcriptional regulation mechanisms. Drug prediction analysis identified atorvastatin as a promising therapeutic candidate with strong binding affinity to multiple key NET-related proteins. Conclusion: Our study provides insights into the temporal dynamics and molecular mechanisms of NET formation after SCI, identifying potential therapeutic targets to mitigate neutrophil-mediated secondary injury and improve functional outcomes.

Source: Journal of Inflammation Research

DOI: 10.2147/jir.s530446