Objectives

The mechanisms underlying the role of mast cells in wound healing have not been thoroughly studied, and even fewer data are available on studies related to mast cells in the skin of patients with type 2 diabetes mellitus (T2DM). Therefore, this study aims to explore the transcriptional characteristics of mast cells at the single-cell level in patients with T2DM and provide experimental data for studying mast cell behaviors under abnormal glucose metabolism.

Methods

Two patients with T2DM and one trauma patient without diabetes were enrolled. Samples were derived from skin tissue resected at the time of surgery and were isolated by single cell capture technology on BD platform to prepare single cell cDNA library. Seurat was used to process raw reads and analyze data downstream of single-cell RNA sequencing, including removal of low-quality cells, identification of cell clusters at the single-cell level, and screening for differential genes with fold change > 1.5 and p < 0.05 by two-sided t-test. We performed single-cell RNA sequencing on skin tissues of T2DM patients and non-diabetics and identified the cell cluster of skin, single-cell subsets, and transcriptional characteristics of mast cells at a single-cell level. Meanwhile, gene set enrichment(GSEA) analysis was performed on the differentially expressed genes.

Results

A total of 8888 cells were obtained from skin tissue. Clustering analysis revealed eight-cell clusters, identified as smooth muscle cells, dendritic cells, mast cells, and T cells, respectively. Cluster 6 was identified as mast cells with the marker genes TPSAB1, CPA3, TPSB2, MS4A2,KIT, etc., which accounting for 2.7% of the total cell number.Compared with the control group, the genes highly expressed in MCs from T2DM patients, include ADH1C, PAXIP1, HAS1, ARG1, etc., and the low expression genes include PHACTR2, GGA1, RASSF2, etc. GSEA analysis suggested that the signal pathways of MCS in T2DM patients included VEGF signaling pathway, Fc gamma R-mediated phagocytosis, the B cell receptor signaling pathway, natural killer cell-mediated cytotoxicity.

Conclusions

The characteristic genes of MCs in the skin tissues of T2DM patients were described at the single-cell level. These genes and enriched signaling pathways provide a theoretical basis and data support for further researches on dermatopathy in patients with diabetes mellitus.

中文翻译：

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2型糖尿病患者皮肤组织来源肥大细胞单细胞转录特征

目标

肥大细胞在伤口愈合中的作用机制尚未得到彻底研究，关于 2 型糖尿病 (T2DM) 患者皮肤中肥大细胞相关研究的数据更少。因此，本研究旨在探索T2DM患者单细胞水平的肥大细胞转录特征，为研究糖代谢异常下的肥大细胞行为提供实验数据。

方法

招募了两名患有 T2DM 的患者和一名没有糖尿病的创伤患者。样本取自手术时切除的皮肤组织，在BD平台上通过单细胞捕获技术进行分离，制备单细胞cDNA文库。Seurat 用于处理单细胞 RNA 测序下游的原始 reads 和分析数据，包括去除低质量细胞，在单细胞水平识别细胞簇，筛选倍数变化 > 1.5 和p  <的差异基因0.05 由两侧t-测试。我们对 T2DM 患者和非糖尿病患者的皮肤组织进行了单细胞 RNA 测序，并在单细胞水平上鉴定了皮肤细胞簇、单细胞亚群和肥大细胞的转录特征。同时，对差异表达的基因进行基因集富集（GSEA）分析。

结果

从皮肤组织中总共获得了 8888 个细胞。聚类分析揭示了八个细胞簇，分别被确定为平滑肌细胞、树突细胞、肥大细胞和 T 细胞。Cluster 6被鉴定为肥大细胞，其标志基因TPSAB1、CPA3、TPSB2、MS4A2、KIT等占总细胞数的2.7%。与对照组相比，T2DM患者MCs中的基因高表达，包括ADH1C、PAXIP1、HAS1、ARG1等，低表达基因包括PHACTR2、GGA1、RASSF2GSEA分析提示T2DM患者MCS信号通路包括VEGF信号通路、FcγR介导的吞噬作用、B细胞受体信号通路、自然杀伤细胞介导的细胞毒作用。

结论

在单细胞水平上描述了 T2DM 患者皮肤组织中 MCs 的特征基因。这些基因和丰富的信号通路为进一步研究糖尿病患者皮肤病提供了理论依据和数据支持。