背景:免疫检查点抑制剂(ICIs)广泛用于癌症治疗;然而,ICI相关性心肌炎(ICI-MC)的出现是一种严重且可能致命的并发症,其病理生理机制尚不清楚.本研究旨在鉴定ICI-MC中关键的免疫相关基因,并利用生物信息学揭示潜在的治疗靶点。
方法:使用GSE180045数据集,其中包括三组-A组:无免疫不良事件的ICI患者,B组:ICI患者发生非心肌炎免疫不良事件,和C组:患有心肌炎的ICI患者-我们分析了ICI-MC样本(C组)和非心肌炎对照组(A组和B组)之间的差异表达基因(DEGs)。然后将这些DEGs与来自immPort数据库的1796个免疫相关基因交叉引用以鉴定免疫相关的DEGs。我们进行了功能富集分析(基因本体论,京都基因和基因组百科全书,基因集富集分析),构建了一个蛋白质-蛋白质相互作用网络,并确定了集线器基因。使用GSE4172数据集的验证导致从hub基因和DEG之间的重叠中鉴定出最佳特征基因。预测目标微小RNA(miRNA),构建了竞争性内源RNA(ceRNA)网络。使用ConnectivityMap数据库预测hub基因的靶药物。
结果:我们确定了ICI-MC和对照组之间的58个DEG,与1796个免疫相关基因相交后,鉴定出32个免疫相关的DEGs。功能分析显示细胞裂解富集,CD8+T细胞受体,自然杀伤细胞介导的细胞毒性,和愤怒的信号。值得注意的是上调的hub基因包括IL7R,PRF1,GNLY,CD3G,NKG7,GZMH,GZMB,KLRB1、KLRK1和CD247。在验证数据集中,407个DEG被发现,从而鉴定出3个最佳特征基因(KLRB1、NKG7、GZMH)。预测的靶miRNA,lincRNAs,和circRNAs构成了一个全面的ceRNA网络。在连接评分升高的前10名药物中,乙酰羟肟酸,表明ICI治疗需要谨慎。
结论:KG7,GZMH,和KLRB1被鉴定为ICI-MC中关键的免疫相关基因。生物富集包括参与细胞裂解的途径,CD8+T细胞受体途径,自然杀伤细胞介导的细胞毒性,RAGE信令,和促炎反应。ceRNA网络阐明了关键分子的作用,并强调了在ICI治疗中避免使用乙酰羟肟酸等药物的重要性。关键信息关于本主题的已知心肌炎被认为是严重的ICI相关毒性,看似罕见,但经常暴发性和致命性。ICI相关心肌炎的潜在机制仍未完全了解。尽管T细胞和细胞毒性T淋巴细胞相关蛋白4(CTLA-4)的意义是显而易见的,煽动抗原,他们认可的原因,导致心肌细胞损伤的机制还没有得到很好的表征。对ICI相关心肌炎的更好理解将提供对免疫系统和心血管系统之间平衡的见解。我们的研究进一步验证了T细胞和CTLA-4在ICI相关心肌炎中的意义。更重要的是,我们确定了三个基因-NKG7,GZMH,和KLRB1对于ICI-MC的开发至关重要,并提出了涉及这三个关键基因的ceRNA网络。这项研究如何影响研究,实践或政策新发现的关键基因及其复杂的分子相互作用为ICI-MC的潜在机制提供了全面的视角。此外,我们的研究结果建议在ICI治疗期间谨慎使用乙酰异羟肟酸等药物.随着我们对这些监管网络的理解加深,我们的研究提供了有价值的见解,可以为ICI-MC的未来治疗策略提供参考.
BACKGROUND: Immune checkpoint inhibitors (ICIs) are widely used in cancer treatment; however, the emergence of ICI-associated myocarditis (ICI-MC) presents a severe and potentially fatal complication with poorly understood pathophysiological mechanisms. This study aimed to identify crucial immune-related genes in ICI-MC and uncover potential therapeutic targets using bioinformatics.
METHODS: Using the GSE180045 dataset, which includes three groups-Group A: ICI patients without immune adverse events, Group B: ICI patients with non-myocarditis immune adverse events, and Group C: ICI patients with myocarditis-we analyzed differentially expressed genes (DEGs) between ICI-MC samples (Group C) and non-myocarditis controls (Groups A and B). These DEGs were then cross-referenced with 1796 immune-related genes from the immPort database to identify immune-related DEGs. We conducted functional enrichment analyses (Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, gene set enrichment analysis), constructed a protein-protein interaction network, and identified hub genes. Validation using the GSE4172 dataset led to the identification of optimal feature genes from the overlap between hub genes and DEGs. Predictions of target MicroRNAs (miRNAs) were made, and a competing endogenous RNA (ceRNA) network was constructed. Target drugs for hub genes were predicted using the Connectivity Map database.
RESULTS: We identified 58 DEGs between ICI-MC and controls, which led to the identification of 32 immune-related DEGs after intersection with 1796 immune-related genes. Functional analyses revealed enrichment in cell lysis, CD8+ T-cell receptor, natural killer cell-mediated cytotoxicity, and RAGE signaling. Notably upregulated hub genes included IL7R, PRF1, GNLY, CD3G, NKG7, GZMH, GZMB, KLRB1, KLRK1, and CD247. In the validation dataset, 407 DEGs were uncovered, resulting in the identification of 3 optimal feature genes (KLRB1, NKG7, GZMH). The predicted target miRNAs, lincRNAs, and circRNAs constituted a comprehensive ceRNA network. Among the top 10 drugs with elevated connectivity scores was acetohydroxamic acid, indicating a need for caution in ICI treatment.
CONCLUSIONS: KG7, GZMH, and KLRB1 were identified as pivotal immune-related genes in ICI-MC. Biological enrichments included pathways involved in cell lysis, the CD8+ T-cell receptor pathway, natural killer cell-mediated cytotoxicity, RAGE signaling, and proinflammatory responses. The ceRNA network illuminated the role of critical molecules and underscored the importance of avoiding drugs such as acetohydroxamic acid in ICI treatment. Key message What is already known on this topic Myocarditis is recognized as a serious ICI-associated toxicity, seemingly infrequent yet often fulminant and lethal. The underlying mechanisms of ICI-associated myocarditis remain not fully understood. Although the significance of T cells and cytotoxic T lymphocyte-associated protein 4 (CTLA-4) is evident, the inciting antigens, the reasons for their recognition, and the mechanisms causing cardiac cell injury are not well characterized. An improved understanding of ICI-associated myocarditis will provide insights into the equilibrium between the immune and cardiovascular systems. What this study adds Our study further validates the significance of T cells and CTLA-4 in ICI-associated myocarditis. More importantly, we identified three genes-NKG7, GZMH, and KLRB1-essential for the development of ICI-MC and proposed ceRNA networks involving these three key genes. How this study might affect research, practice or policy The newly discovered key genes and their intricate molecular interactions offer a comprehensive perspective on the mechanisms underlying ICI-MC. Furthermore, our findings advise caution regarding the use of drugs like acetohydroxamic acid during ICI treatment. As our understanding of these regulatory networks deepens, our study provides valuable insights that could inform future therapeutic strategies for ICI-MC.