Neurotrophin-3/chitosan inhibits cuproptosis-related genes to enable functional recovery after spinal cord injury.

Abstract

OBJECTIVES: This study investigated the regulatory mechanisms of cuproptosis-related genes (CRGs) in spinal cord injury (SCI) and explored the therapeutic potential of neurotrophin-3 (NT3)-loaded chitosan in promoting functional recovery. METHODS: We conducted integrated bulk RNA-seq and single-cell RNA-seq (scRNA-seq) analyses of mouse spinal cord tissue at various time points after SCI. The key CRGs were identified using differential expression analysis, weighted gene co-expression network analysis, and machine learning. The therapeutic effects of NT3-loaded chitosan were evaluated using animal models and molecular docking analysis. RESULTS: We identified four key CRGs (Atp7a, Cp, Loxl2, and Pde3b) and three key transcription factors (C/EBPα, Stat6, and Runx1) that were upregulated post-SCI, promoting cuproptosis and neuroinflammation. NT3-loaded chitosan treatment significantly inhibited CRG expression and enhanced functional recovery in the animal models. Molecular docking analysis demonstrated binding interactions between chitosan and key CRGs, suggesting a potential mechanism for their therapeutic effects. CONCLUSIONS: Our findings highlight the critical role of CRGs in SCI progression and the potential of NT3-loaded chitosan as a therapeutic strategy for inhibiting cuproptosis and promoting functional recovery. Future studies should focus on validating these findings in larger cohorts and exploring the detailed mechanisms by which NT3-loaded chitosan modulates CRG expression.