摘要
背景：甲状腺眼病是一种病因复杂的慢性炎症性自身免疫性疾病。N6-甲基腺苷（m6A）修饰和自噬分别被证实参与了TED过程。越来越多的证据表明，m6A在各种疾病中调节自噬至关重要。然而，关于m6A修饰和自噬在TED中的相互作用的知识有限。我们的研究旨在研究m6A修饰和自噬相互作用在TED中的作用。
结果：与健康对照组相比，我们首次发现TED患者中5种m6A调节因子和44种ARG的失调。经过相关性分析、与RM2靶点数据库中已验证的靶基因重叠以及在我们的样本中进行验证，FTO和BNIP3被认为是TED的生物标志物。随后，我们分别基于失调的m6A调节因子和ARGs，将27名TED患者分为两个集群，并进一步评估集群的免疫浸润特征。鉴定了聚类相关的差异表达基因，并阐明了其丰富的生物学功能和途径。此外，使用两种机器学习算法，我们为具有不同分子簇的TED患者创建了一个预测模型。进行列线图、校准曲线和决策曲线分析，以评估预测模型的性能。
结论：本研究表明，FTO和BNIP3之间的相互作用，抑制的FTO可能以m6A依赖的方式下调BNIP3的表达，抑制自噬，随后促进TED过程。此外，我们还构建了一个用于预测TED的列线图模型。这些结果为理解TED的机制提供了新的见解。
Abstract
Background:Thyroid eye disease (TED) is a chronic inflammatory autoimmune disease with a complex etiology. N6-methyladenosine (m6A) modification and autophagy were confirmed separately to be involved in the TED process. Increased evidence has shown that m6A is critical in regulating autophagy in various diseases. However, there is limited knowledge about the interactive effects of m6A modification and autophagy in TED. Our research aimed to investigate the effects of m6A modification and autophagy interactivity in TED.

Results: We first identified dysregulation of five m6A regulators and 44 ARGs in TED patients compared to healthy controls. After correlation analysis, overlapping with the validated target genes from the RM2target database, and verification in our samples, FTO and BNIP3 were considered biomarkers for TED. Subsequently, based on dysregulated m6A regulators and ARGs separately, we classified 27 TED patients into two clusters, and the immune infiltration characteristics of clusters were further evaluated. Cluster-related differentially expressed genes were identified, and the enriched biological functions and pathways were elucidated. In addition, using two machine learning algorithms, we created a prediction model for TED patients with different molecular clusters. The nomogram, calibration curve, and decision curve analysis were performed to assess the performance of the predictive model.

Conclusions: This study revealed that an interactive effect between FTO and BNIP3, suppressed FTO might downregulate the expression of BNIP3 in an m6A-dependent manner, inhibiting the autophagy and subsequently promoting the TED process. In addition, we constructed a nomogram model in predicting the TED. These results provide new insights into understanding the mechanism of TED.

https://www.researchsquare.com/article/rs-2759942/v1