Cao Liang, Li Qing
Department of Traumatic Orthopedics, The Second Affiliated Hospital, University of South China, Hengyang, China.
School of Clinical Medicine, Guizhou Medical University, Guiyang, China.
Front Genet. 2022 May 30;13:883810. doi: 10.3389/fgene.2022.883810. eCollection 2022.
Spinal cord injury (SCI) is a disabling condition with significant morbidity and mortality. Currently, no effective SCI treatment exists. This study aimed to identify potential biomarkers and characterize the properties of immune cell infiltration during this pathological event. To eliminate batch effects, we concurrently analyzed two mouse SCI datasets (GSE5296, GSE47681) from the GEO database. First, we identified differentially expressed genes (DEGs) using linear models for microarray data (LIMMA) and performed functional enrichment studies on those DEGs. Next, we employed bioinformatics and machine-learning methods to identify and define the characteristic genes of SCI. Finally, we validated them using immunofluorescence and qRT-PCR. Additionally, this study assessed the inflammatory status of SCI by identifying cell types using CIBERSORT. Furthermore, we investigated the link between key markers and infiltrating immune cells. In total, we identified 561 robust DEGs. We identified Rab20 and Klf6 as SCI-specific biomarkers and demonstrated their significance using qRT-PCR in the mouse model. According to the examination of immune cell infiltration, M0, M1, and M2 macrophages, along with naive CD8, dendritic cell-activated, and CD4 Follicular T cells may have a role in the progression of SCI. Therefore, Rab20 and Klf6 could be accessible targets for diagnosing and treating SCI. Moreover, as previously stated, immune cell infiltration may significantly impact the development and progression of SCI.
脊髓损伤(SCI)是一种致残性疾病,具有较高的发病率和死亡率。目前,尚无有效的脊髓损伤治疗方法。本研究旨在识别潜在的生物标志物,并描述这一病理过程中免疫细胞浸润的特性。为消除批次效应,我们同时分析了来自基因表达综合数据库(GEO)的两个小鼠脊髓损伤数据集(GSE5296、GSE47681)。首先,我们使用微阵列数据线性模型(LIMMA)识别差异表达基因(DEG),并对这些差异表达基因进行功能富集研究。接下来,我们采用生物信息学和机器学习方法来识别和定义脊髓损伤的特征基因。最后,我们通过免疫荧光和定量逆转录聚合酶链反应(qRT-PCR)对其进行验证。此外,本研究通过使用CIBERSORT识别细胞类型来评估脊髓损伤的炎症状态。此外,我们还研究了关键标志物与浸润免疫细胞之间的联系。我们总共识别出561个稳定的差异表达基因。我们将Rab20和Klf6鉴定为脊髓损伤特异性生物标志物,并在小鼠模型中通过qRT-PCR证明了它们的重要性。根据对免疫细胞浸润的检测,M0、M1和M2巨噬细胞,以及幼稚CD8、活化树突状细胞和CD4滤泡辅助性T细胞可能在脊髓损伤的进展中起作用。因此,Rab20和Klf6可能是诊断和治疗脊髓损伤的可及靶点。此外,如前所述,免疫细胞浸润可能会显著影响脊髓损伤的发生和发展。