基于m6A调节因子的综合分析鉴定溃疡性结肠炎的生物标志物及预测治疗药物

Identification of biomarkers of ulcerative colitis and prediction of therapeutic drugs based on comprehensive analysis of m6A regulators

关键词:  m6A  溃疡性结肠炎  免疫浸润  生物标志物  治疗靶点

Key words:  m6A  ulcerative colitis  immune infiltration  biomarker  therapeutic target

摘要:

摘要：目的 阐释N6甲基化腺苷(N6-methyl-adenosine，m6A)甲基化调节因子在溃疡性结肠炎(ulcerative colitis，UC)中的作用，探讨m6A与UC免疫调节的关系，寻找UC的治疗靶点及药物或协助临床决策的生物标志物。方法 从GEO数据库下载GSE87466数据集，筛选差异表达的m6A调节因子并进行动物实验验证。采用随机森林模型筛选3个m6A调节因子作为特征基因。建立列线图模型预测UC的患病风险。采用共识聚类方法对UC样本进行分类。使用ssGSEA分析评估样本中免疫细胞的丰度，量化免疫浸润。使用Coremine Medical数据库、HERB数据库及CTD数据库预测治疗UC的潜在中药及中药成份并进行分子对接验证。结果 与正常组织相比，UC组织中METTL3、HNRNPA2B1、IGF2BP1、YTHDF3和LRPPRC的表达水平显著不同。GO富集分析表明，m6A调节因子与炎症免疫有关。基于LRPPRC、YTHDF3及IGF2BP1三个特征基因构建的列线图模型具有良好的预测能力。免疫浸润分析发现22种免疫细胞的浸润丰度均显著高于正常组织样本。聚类分析显示UC中具有两种不同的m6A修饰模式，它们具有不同的免疫浸润特征和细胞因子水平。雷公藤、长春新碱及雷公藤内酯酮等与差异表达的m6A调节因子的关系最为密切，分子对接结果显示它们之间具有较好的结合活性。结论 m6A调节因子的异常表达与UC的发生发展密切相关，LRPPRC、IGF2BP1及YTHDF3有望成为诊断和治疗UC的生物标志物。此外，雷公藤、长春新碱及雷公藤内酯酮等中药或中药成份有望成为潜在的抗UC药物。该研究为UC的临床治疗和诊断提供了新的线索，为抗UC新药的开发提供参考方向。

Abstract:

ABSTRACT: OBJECTIVE Explain the role of N6 methyladenosine (m6A) methylation regulators in ulcerative colitis (UC), and explore the relationship between m6A and UC immune regulation. Search for therapeutic targets and drugs of UC or biomarkers to assist clinical decision-making. METHODS The GSE87466 data set was downloaded from the GEO database, and the differentially expressed m6A regulators were screened and verified by animal experiments. Random forest model was used to screen three m6A regulators as characteristic genes. A nomograph model was established to predict the risk of UC. Consensus clustering method is used to classify UC samples. ssGSEA analysis was used to evaluate the abundance of immune cells in samples and quantify immune infiltration. The coremine medical database, HERB database and CTD database were used to predict the traditional Chinese medicine and its components for the treatment of UC and carry out molecular docking verification. RESULTS Compared with normal tissues, the expression levels of METTL3, HNRNPA2B1, IGF2BP1, YTHDF3 and LRPPRC in UC tissues were significantly different. GO enrichment analysis showed that m6A regulators were related to inflammatory immunity. The nomogram model based on LRPPRC, YTHDF3 and IGF2BP1 has good prediction ability. Immune infiltration analysis showed that the infiltration abundance of the remaining 22 immune cells in UC samples were significantly higher than that in normal samples. Cluster analysis showed that there were two different m6A modification patterns in UC, which had different immune infiltration characteristics and cytokine levels. Tripterygium wilfordii, vincristine and triptonide are most closely related to the differentially expressed m6A regulators. Molecular docking results show that they have good binding activity. CONCLUSION The differential expression of m6A regulators is closely related to the occurrence and development of UC. LRPPRC, YTHDF3 and IGF2BP1 are expected to become biomarkers for the diagnosis and treatment of UC. Tripterygium wilfordii, vincristine, triptonide and other traditional Chinese medicine (TCM) or TCM components are expected to become potential drugs for the treatment of UC. This study provides new clues for clinical treatment and diagnosis and a reference direction for the development of new drugs against UC.