缺血性脑卒中患者双硫死亡相关基因的差异表达及其与免疫调节的关联研究

Study on the differential expression of disulfide death-related genes and their association with immune regulation in patients with ischemic stroke

摘要:

目的]缺血性脑卒中(IS)是由于脑血管局部急性缺血诱发的脑组织损伤与神经元凋亡,其发病机制复杂,涉及细胞焦亡、铁死亡和双硫死亡等多种细胞死亡方式。双硫死亡是一种新近发现的死亡形式,有助于从新的视角探究各种疾病的病理机制。本研究旨在发现与验证缺血患者血液样本中双硫死亡相关基因的差异表达及其与免疫调节的关联。 [方法]通过网络大数据获得临床患者的相关数据集(GSE16561和GSE37587),基于该数据集对双硫死亡相关基因进行差异分析和基因富集分析。随后对免疫细胞浸润进行分析,以研究IS背景下免疫细胞的失调。最后通过ROC曲线、列线图、校准曲线和决策曲线等验证关键基因的准确性和构建疾病预测模型以预测脑卒中患病的风险。 [结果]基于GSE16561、GSE37587数据集确定了9个双硫死亡相关基因的显著差异表达。外部数据集GSE58294验证显示,在69例IS患者和23例正常对照者血液样本中,LRPPRC、MYH9、NDUFA11、PRDX1和RPN1这5个差异表达基因的趋势一致。免疫浸润分析发现,TLN1、MYH9、PRDX1、LRPPRC、NDUFA11等差异基因与IS患者CD8＋T细胞、活化NK细胞、巨噬细胞、中性粒细胞有强相关性。功能富集分析显示,局灶性黏附、血小板聚集与活化等途径在IS疾病的发生发展中有重要作用。线图模型风险预测表明,这些差异基因准确性良好,双硫死亡相关基因优化的ROC曲线AUC值可达0.844。外部数据集GSE58294验证发现,双硫死亡相关基因优化的ROC曲线AUC值可达0.989,对IS的患病风险有良好的临床指导意义。 [结论]本研究通过数据集证实了IS患者中存在5个双硫死亡相关基因,其中MYH9上调,LRPPRC、NDUFA11、PRDX1和RPN1下调,这些基因的变化可能从免疫炎症、出血风险等方面影响IS疾病进展及预后。

Abstract:

Aim Ischemic stroke (IS) is caused by acute ischemia of cerebral blood vessels, leading to brain tissue damage and neuronal apoptosis. The pathogenesis is complex, involving multiple cell death modes such as pyroptosis, ferroptosis and disulfide death. Disulfide death is a newly discovered form of death that helps to explore the pathological mechanisms of various diseases from a new perspective. The aim of this study is to discover and validate the differential expression of disulfide death-related genes in blood samples of ischemic patients and their association with immune regulation. Methods The relevant datasets of clinical patients (GSE16561 and GSE37587) were obtained through online big data. Differentially expressed genes related to disulfide death were identified, and gene enrichment analysis was conducted to further explore the potential mechanisms. Subsequently, immune cell infiltration was analyzed to investigate the dysregulation of immune cells in the context of IS. Finally, the accuracy of key genes was verified through ROC curves, column charts, calibration curves, and decision curves, and a disease prediction model was constructed to predict the risk of stroke. Results Based on this dataset, significant differential expression of 9 genes related to disulfide death was identified. Independent external validation was conducted using the microarray dataset GSE58294. Single item comparisons were performed on these differentially expressed genes in blood samples from 69 IS patients and 23 normal individuals.The results showed that the trends of LRPPRC, MYH9, NDUFA11, PRDX1 and RPN1, the 5 differentially expressed genes, were consistent. Immune infiltration analysis found that differentially expressed genes such as TLN1, MYH9, PRDX1, LRPPRC, NDUFA11 were also strongly correlated with CD8＋T cells, activated NK cells, macrophages, and neutrophils in IS patients. Functional enrichment analysis emphasized the important role of pathways such as focal adhesion, platelet aggregation, and activation in the occurrence and development of diseases. By using a column chart model for risk prediction, it was shown that the accuracy of these differentially expressed genes was good, and the ROC curve AUC value of the optimized combination of disulfide death-related genes could reach 0.844. Further validation through an external dataset (GSE58294) revealed that the ROC curve AUC value optimized for disulfide death-related genes reached 0.989, which had good clinical guidance significance for the risk of IS. Conclusions This study confirmed the existence of 5 disulfide death-related genes in IS patients through a dataset, including upregulation of MYH9 and downregulation of LRPPRC, NDUFA11, PRDX1 and RPN1. These gene alterations are suggested to influence IS disease progression and prognosis through immune inflammation and bleeding risk.