Mao Dandan, Chen Qi, Tong Shuolan, Xu Zixia, Yu Guofeng, Chang Chuan, Lv Yao
Department of Neurosurgery, The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Quzhou, China.
Department of Nursing, Huashan Hospital, Fudan University, Shanghai, China.
Sci Rep. 2025 Jan 15;15(1):1993. doi: 10.1038/s41598-025-86170-0.
Spinal cord injury (SCI) is a profound ailment lacking a well-defined molecular mechanism and effective treatments. Cuproptosis, identified as a recently discovered cell death pathway, exhibits diverse roles in various cancers. Nevertheless, its involvement in SCI is yet to be elucidated. Firstly, the RNA sequencing data of 1, 3, 7 dpi SCI samples were collected from GEO database. We performed differential expression analysis on these samples with varying cuproptosis-related scores calculating by ssGSEA. Subsequently, we conducted enrichment analyses with KEGG, GO, and GSEA. Simultaneously, we executed WGCNA analysis using cuproptosis-related scores, selecting the most relevant module for enrichment analysis. Hub genes were identified at the intersection of PPI analysis results from two modules and cuproptosis-related DEGs. Additionally, relying on the immune infiltration landscape associated with cuproptosis, we carried out immune cell correlation analysis on hub genes. Finally, to corroborate our earlier findings, we utilized single-cell RNA-seq analysis and in vitro experimental validation. Based on ssGSEA, differential expression analysis and WGCNA analysis, we identified two modules that were highly relevant to cell division and immune processes, respectively. From these modules, we identified two hub genes, Cd48 and Mpeg1, which exhibited a strong positive correlation (R = 0.92) and shared similar pathways. Furthermore, we observed a positive correlation between M2 macrophages and Cd48/Mpeg1. To validate our findings, we performed external cohort validation using a single-cell RNA sequencing dataset. The results confirmed that Mpeg1 was highly expressed in microglia (macrophages in center nervous system) following spinal cord injury. Additionally, we conducted in vitro experiments to further validate the molecular functions of Mpeg1 in SCI. In summary, targeting Mpeg1, as well as cuproptosis and immune cell infiltration, holds promise as a potential strategy for reducing spinal cord tissue damage and promoting recovery after SCI. These findings provide valuable insights for future therapeutic interventions.
脊髓损伤（SCI）是一种严重疾病，缺乏明确的分子机制和有效治疗方法。铜死亡作为最近发现的一种细胞死亡途径，在各种癌症中发挥着多种作用。然而，其在脊髓损伤中的作用尚待阐明。首先，从基因表达综合数据库（GEO数据库）收集1、3、7天脊髓损伤样本的RNA测序数据。我们对这些样本进行差异表达分析，通过单样本基因集富集分析（ssGSEA）计算不同的铜死亡相关评分。随后，我们进行了京都基因与基因组百科全书（KEGG）、基因本体（GO）和基因集富集分析（GSEA）。同时，我们使用铜死亡相关评分进行加权基因共表达网络分析（WGCNA），选择最相关的模块进行富集分析。在两个模块的蛋白质-蛋白质相互作用（PPI）分析结果与铜死亡相关的差异表达基因（DEGs）的交集处鉴定出枢纽基因。此外，基于与铜死亡相关的免疫浸润图谱，我们对枢纽基因进行了免疫细胞相关性分析。最后，为了证实我们早期的发现，我们利用单细胞RNA测序分析和体外实验验证。基于ssGSEA、差异表达分析和WGCNA分析，我们确定了分别与细胞分裂和免疫过程高度相关的两个模块。从这些模块中，我们鉴定出两个枢纽基因，Cd48和Mpeg1，它们呈现出强正相关（R = 0.92）且共享相似的途径。此外，我们观察到M2巨噬细胞与Cd48/Mpeg1之间存在正相关。为了验证我们的发现，我们使用单细胞RNA测序数据集进行了外部队列验证。结果证实，脊髓损伤后Mpeg1在小胶质细胞（中枢神经系统中的巨噬细胞）中高表达。此外，我们进行了体外实验以进一步验证Mpeg1在脊髓损伤中的分子功能。总之，靶向Mpeg1以及铜死亡和免疫细胞浸润，有望成为减少脊髓组织损伤和促进脊髓损伤后恢复的潜在策略。这些发现为未来的治疗干预提供了有价值的见解。
