Characteristic analysis of skin keratinocytes in patients with type 2 diabetes based on the single-cell levels

Abstract

Background: Keratinocytes play an important role in wound healing; however, less is known about skin keratinocytes in patients with type 2 diabetes mellitus (T2DM). Therefore, this study aimed to search for the transcriptional characteristics of keratinocytes at the single-cell level from T2DM patients, and to provide experimental data for identifying the pathological mechanisms of keratinocytes under pathological conditions. Methods: We performed single-cell RNA sequencing on the skin tissue from two T2DM patients and one patient without diabetes-induced trauma using the BD Rhapsody™ Single-Cell Analysis System. With the help of bioinformatics R-based single-cell analysis software, we analyzed the results of single-cell sequencing to identify the single-cell subsets and transcriptional characteristics of keratinocytes at the single-cell level, including Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyzes. Results: In this study, we found specific highly expressed signature keratinocyte-related genes. We analyzed the transcriptome of keratinocytes from experimental and control groups and screened a total of 356 differential genes, which were subject to bioinformatics analysis. Enriched pathways included oxidative phosphorylation, antigen processing and presentation, prion and Huntingtons’ diseases, bacterial invasion of epithelial cells, thermogenesis, vasopressin-regulated water reabsorption, and protein processing in the endoplasmic reticulum. Conclusions: This study revealed the characteristics of keratinocytes at the single-cell level and screened a group of differentially expressed genes related to T2DM-associated keratinocytes, oxidative phosphorylation, cytokine receptor interactions, prion diseases, and other signaling pathways.

Cellular Patterns of the Skin Microenvironment in Experimentally Induced Diabetes Mellitus

“…Collagen fi-bres are thick, long and with sinuous tracts, forming a dense, semi-coordinated connective tissue (Figure 7). In the presence of chronic hyperglycaemia, free glucose molecules spon-taneously attach to protein molecules in a process of non-enzymatic glycosylation, which has also been shown in other proteins, which leads to morphological and molecular changes that significantly influence some physical properties, such as flexibility [6][7][8][9]24].Early characterization and quantification of the pathophysiological details of diabetes mellitus and its complications correlates with more effective diagnostic and therapeutic methods, and implicitly with improved survival and quality of life [23,[37][38][39][40].…”

Single-cell sequencing technology in diabetic wound healing: New insights into the progenitors-based repair strategies

“…Additionally, the proportion of M1 macrophages (classically activated macrophages that promote inflammation) was higher in DFU-healer than in DFU-non-healers, compared to that of M2 macrophages (alternatively activated macrophages with anti-inflammatory properties)[ 9 ].These results implied that suppressing systemic immuno-inflammatory responses while activating local responses in the wound environment could facilitate diabetic wound healing. Liao et al [ 10 ] found highly expressed keratinocyte genes in the diabetic wound, including SFN, LYPD3, S100A8, KRT1, KRT10, KRT6A, KRT5, and KRT16, underscoring the crucial role of keratinocytes in diabetic wound healing. By analysing the skin specimens of DFU patients and healthy controls using scRNAseq, Du et al [ 11 ] found that human dermal microvascular endothelial cells (HDMECs) isolated from DFU patients showed considerably impaired tube formation compared to those from healthy controls; they also found that the significantly under-expressed RAB17 in DFU-HDMECs may be the key factor leading to the impaired angiogenic capacity in DFUs.…”

“…The differential gene expression revealed by scRNAseq also suggested the activation or inhibition of the corresponding signalling pathway in the process of diabetic wound healing. The diabetic healing-related differentially expressed gene analysis and gene ontology functional enrichment analysis identified significant differential genes, including CD19, Integrin Subunit Alpha M, HLA-DR, CXC chemokine ligand 11, MMP1, heparan sulfate 3-O-sulfotransferase 2, CALML, interleukin (IL)7R, IL6, TCF7, CCR7, IL1B, S100A8, HIF1α, TNF, CD44, transforming growth factor β1, C-C chemokine ligand 5, SOX4, RAB17, CD200 and vascular endothelial growth factor A[ 7 , 10 ]. These genes were predominantly associated with the immune and inflammatory signalling pathways, oxidative phosphorylation and cytokine receptor interactions, suggesting that the immune and inflammatory environment is critical for diabetic wound healing.…”

“…These genes were predominantly associated with the immune and inflammatory signalling pathways, oxidative phosphorylation and cytokine receptor interactions, suggesting that the immune and inflammatory environment is critical for diabetic wound healing. Alterations in the metabolic processes of cells within diabetic wounds have also been implied[ 7 , 10 ]. Additionally, genes such as ANPEP, BID, CYBA, CYBB, FCER1G, ITGA1 and PLAUR, which were overexpressed in the diabetic wound microenvironment, might serve as potential drug targets[ 7 ].…”