背景:通过鉴定与DR中的角化相关的分子生物学标记,可以开发新的病理生物学途径和更容易获得的诊断标志物。
方法:与糖尿病视网膜病变(DR)相关的数据集从基因表达综合数据库获得,而与角化相关的基因来自先前发表的汇编。进行共识聚类以描绘不同的DR子类。利用加权相关网络分析(WGCNA)鉴定特征基因。此外,采用两种机器学习算法来优化特征基因的选择.最后,我们使用链脲佐菌素诱导的糖尿病小鼠模型和高糖诱导的BV2模型,进行了初步验证实验,以确定角化在DR发生和关键基因转录调控中的作用.
结果:在STZ诱导的糖尿病小鼠视网膜中,角化标记蛋白表达的减少(FDX1,DLAT,和NDUFS8)提示DR发生角化。随后,通过STZ诱导的糖尿病和氧诱导的视网膜病变(OIR)模型验证了八个角化差异基因的表达,关键基因SLC31A1在模型和数据集物种中均显示出上调。进一步分析,包括加权基因共表达网络分析,GSVA,和免疫浸润分析,表明角化凋亡与小胶质细胞功能密切相关。此外,在小胶质细胞体外模型中的验证表明在高葡萄糖条件下小胶质细胞中发生了角化现象,STAT1异常表达与SLC31A1。
结论:我们的研究结果表明,STAT1/SLC31A1可能为更深入地理解DR的机制基础铺平道路,并提供潜在的治疗途径。
BACKGROUND: By identifying molecular biological markers linked to
cuproptosis in diabetic retinopathy (DR), new pathobiological pathways and more accessible diagnostic markers can be developed.
METHODS: The datasets related to DR were acquired from the Gene Expression Omnibus database, while genes associated with
cuproptosis were sourced from previously published compilations. Consensus clustering was conducted to delineate distinct DR subclasses. Feature genes were identified utilizing weighted correlation network analysis (WGCNA). Additionally, two machine-learning algorithms were employed to refine the selection of feature genes. Finally, we conducted preliminary validation experiments to ascertain the involvement of
cuproptosis in DR development and the transcriptional regulation of critical genes using both the streptozotocin-induced diabetic mouse model and the high glucose-induced BV2 model.
RESULTS: In the STZ-induced diabetic mouse retinas, a decrease in the expression of cuproptosis signature proteins (FDX1, DLAT, and NDUFS8) suggested the occurrence of
cuproptosis in DR. Subsequently, the expression of eight cuproptosis differential genes was validated through the STZ-induced diabetes and oxygen-induced retinopathy (OIR) models, with the key gene SLC31A1 showing upregulation in both models and dataset species. Further analyses, including weighted gene co-expression network analysis, GSVA, and immune infiltration analysis, indicated a close correlation between cuproptosis and microglia function. Additionally, validation in an in vitro model of microglia indicated the occurrence of
cuproptosis in microglia under high glucose conditions, alongside abnormal expression of STAT1 with SLC31A1.
CONCLUSIONS: Our findings suggest that STAT1/SLC31A1 may pave the way for a deeper comprehension of the mechanistic basis of DR and offer potential therapeutic avenues.