目的:本研究旨在探讨成纤维细胞亚型的独特转录特征以及成纤维细胞凋亡在糖尿病足溃疡(DFU)中的作用。
方法:搜索GEO(基因表达综合)以获得DFU单细胞和转录数据集。在通过经典标记基因识别细胞类型后,整合的单细胞数据集用于运行轨迹推断,RNA速度,和配体-受体相互作用分析。接下来,将DFU的大量RNA-seq数据集分析为关键的铁凋亡基因。
结果:这里,我们利用来自GEO数据库的DFU的单细胞测序(scRNA-seq)数据从足部样本中提取83529个单转录组,并鉴定出12种细胞类型,成纤维细胞表现出铁死亡活性水平升高和大量细胞异质性。我们的结果定义了六个主要的成纤维细胞亚群,显示间充质,分泌型网状,分泌-乳头状,促炎,肌生成,和愈合丰富的功能注释。轨迹推断和细胞-细胞通讯分析揭示了两种主要的细胞命运,具有成纤维细胞亚群和配体-受体相互作用的改变。大量RNA测序数据将CGNL1鉴定为成纤维细胞中的独特诊断特征。值得注意的是,CGNL1与促炎性成纤维细胞呈正相关。
结论:总体而言,我们的分析描绘了DFU细胞群体中存在的异质性,显示出不同的成纤维细胞亚型,其特征在于其独特的转录特征和富集功能。我们的研究将帮助我们更好地了解DFU的发病机制,并将CGNL1确定为DFU治疗的潜在靶标。
OBJECTIVE: This study aimed to explore the unique transcriptional feature of fibroblasts subtypes and the role of ferroptosis in diabetic foot ulcers (DFUs).
METHODS: The GEO (Gene Expression Omnibus) was searched to obtain the DFUs single-cell and transcriptional datasets. After identifying cell types by classic marker genes, the integrated single-cell dataset was used to run trajectory inference, RNA velocity, and ligand-receptor interaction analysis. Next, bulk RNA-seq datasets of DFUs were analyzed to the key ferroptosis genes.
RESULTS: Here, we profile 83529 single transcriptomes from the foot samples utilizing single-cell sequencing (scRNA-seq) data of DFU from GEO database and identified 12 cell types, with fibroblasts exhibiting elevated levels of ferroptosis activity and substantial cellular heterogeneity. Our results defined six main fibroblast subsets that showed mesenchymal, secretory-reticular, secretory-papillary, pro-inflammatory, myogenesis, and healing-enriched functional annotations. Trajectory inference and cell-cell communication analysis revealed two major cell fates with subpopulations of fibroblasts and altered ligand-receptor interactions. Bulk RNA sequencing data identified CGNL1 as a distinctive diagnostic signature in fibroblasts. Notably, CGNL1 positively correlated with pro-inflammatory fibroblasts.
CONCLUSIONS: Overall, our analysis delineated the heterogeneity present in cell populations of DFUs, showing distinct fibroblast subtypes characterized by their own unique transcriptional features and enrichment functions. Our study will help us better understand DFUs pathogenesis and identifies CGNL1 as a potential target for DFUs therapies.