Analysis of the Autophagy-related Gene NLRC4 in spinal cord injury.

Abstract

Background/objectives: To investigate the role of the autophagy-related gene NLR Family CARD Domain Containing 4 (NLRC4) in spinal cord injury via bioinformatics methods, which may provide new targets for the diagnosis and treatment of spinal cord injury.

Methods: This analysis is based on the GEO database dataset GSE151371. To identify potential autophagy-related genes involved in SCI, protein‒protein interaction (PPI) networks were analyzed. Immune microenvironment analysis (LM22) was performed via the CIBERSORTx database to determine the makeup of 22 immune cell types. Furthermore, a rat spinal cord injury model was generated, and the expression of selected autophagy-related genes was validated via immunofluorescence labeling and Western blotting.

Results: Disease enrichment analysis via the Metascape database revealed enrichment for diseases related to the spinal cord, inflammation, infection, and immunity, which aligns with the functional analysis results of previously identified genes. Through the PPI and autophagy-related genes, we identified NLRC4 within the key subnetwork of the PPI network, highlighting its significance as a key signature gene associated with SCI. NLRC4 expression was significantly increased in the three groups, which was correlated with the severity of SCI. In the rat SCI model, NLRC4 protein expression was significantly greater in the SCI group than in the sham group (p < 0.001), confirming the validity of the model.

Conclusions: Since NLRC4 is an important gene involved in the autophagy that leads to spinal cord damage, it can be utilized to illuminate the optimal approach to immunotherapy for individuals with SCI and uncover new targets for therapy.