背景:肺缺血再灌注损伤(IRI)是肺移植后原发性移植物功能障碍(PGD)的主要原因,影响移植后的短期和长期死亡率。全角下垂,一种新发现的涉及凋亡的调节细胞死亡形式,坏死,和焦亡,现在被认为是器官损伤和IRI的可能原因。然而,肺移植后PANoptosis对肺IRI发展的具体作用尚不完全清楚.
方法:在本研究中,我们通过分析GEO数据库中与肺移植后肺IRI相关的基因表达数据,鉴定了差异表达基因(DEGs).根据PANoptosis相关基因的交集确定PANoptosis-IRIDEGs并筛选DEGs。使用Lasso回归和SVM-RFE算法进一步筛选与肺IRI相关的Hub基因。此外,采用Cibersort算法评估免疫细胞浸润,并研究免疫细胞与hub基因之间的相互作用。还分析了可调节hub基因的上游miRNA和可与hub基因相互作用的化合物。此外,利用外部数据集来验证hub基因的差异表达分析.最后,使用定量实时PCR最终确认了hub基因的表达,西方印迹,在肺IRI和肺移植患者的动物模型中和免疫组织化学。
结果:全景相关基因,特别是干扰素调节因子1(IRF1)和白细胞介素1α(IL1A),已被确定为肺移植后肺IRI的潜在生物标志物。在小鼠肺IRI模型中,与对照组相比,IRI组肺组织中IRF1和IL1A的mRNA和蛋白表达水平均显着升高。此外,与健康对照组相比,肺移植受者的PBMC中IRF1和IL1A蛋白水平显著升高.经历过PGD的患者在其血液样本中显示IRF1和IL1A蛋白水平升高。此外,在接受肺移植的患者中,与健康对照组相比,外周血单核细胞(PBMC)中IRF1和IL1A蛋白水平显著升高.此外,发生原发性移植物功能障碍(PGD)的患者比没有发生PGD的患者表现出更高的IRF1和IL1A蛋白水平.此外,在肺IRI的小鼠模型的肺组织和接受肺移植的患者的PBMC中观察到了PANoptosis。
结论:我们的研究确定IRF1和IL1A是与肺IRI中PANoptosis相关的生物标志物,提示它们作为肺移植后诊断和治疗干预肺IRI和PGD的潜在用途。
BACKGROUND: Lung ischemia reperfusion injury (IRI) is the principal cause of primary graft dysfunction (PGD) after lung transplantation, affecting short-term and long-term mortality post-transplantation. PANoptosis, a newly identified form of regulated cell death involving apoptosis, necroptosis, and pyroptosis, is now considered a possible cause of organ damage and IRI. However, the specific role of PANoptosis to the development of lung IRI following lung transplantation is still not fully understood.
METHODS: In this study, we identified differentially expressed genes (DEGs) by analyzing the gene expression data from the GEO database related to lung IRI following lung transplantation. PANoptosis-IRI DEGs were determined based on the intersection of PANoptosis-related genes and screened DEGs. Hub genes associated with lung IRI were further screened using Lasso regression and the SVM-RFE algorithm. Additionally, the Cibersort algorithm was employed to assess immune cell infiltration and investigate the interaction between immune cells and hub genes. The upstream miRNAs that may regulate hub genes and compounds that may interact with hub genes were also analyzed. Moreover, an external dataset was utilized to validate the differential expression analysis of hub genes. Finally, the expressions of hub genes were ultimately confirmed using quantitative real-time PCR, western blotting, and immunohistochemistry in both animal models of lung IRI and lung transplant patients.
RESULTS: PANoptosis-related genes, specifically interferon regulatory factor 1 (IRF1) and interleukin 1 alpha (IL1A), have been identified as potential biomarkers for lung IRI following lung transplantation. In mouse models of lung IRI, both the mRNA and protein expression levels of IRF1 and IL1A were significantly elevated in lung tissues of the IRI group compared to the control group. Moreover, lung transplant recipients exhibited significantly higher protein levels of IRF1 and IL1A in PBMCs when compared to healthy controls. Patients who experienced PGD showed elevated levels of IRF1 and IL1A proteins in their blood samples. Furthermore, in patients undergoing lung transplantation, the protein levels of IRF1 and IL1A were notably increased in peripheral blood mononuclear cells (PBMCs) compared to healthy controls. In addition, patients who developed primary graft dysfunction (PGD) exhibited even higher protein levels of IRF1 and IL1A than those without PGD. Furthermore, PANoptosis was observed in the lung tissues of mouse models of lung IRI and in the PBMCs of patients who underwent lung transplantation.
CONCLUSIONS: Our research identified IRF1 and IL1A as biomarkers associated with PANoptosis in lung IRI, suggesting their potential utility as targets for diagnosing and therapeutically intervening in lung IRI and PGD following lung transplantation.