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Single Cell Analysis of Gene expression profile as single cell level of chronic diabetic foot ulcers and chronic non-diabetic foot ulcer Foot Wounds

ABSTRACT: Single Cell Analysis of Gene expression profile as single cell level of chronic diabetic foot ulcers and chronic non-diabetic foot ulcer Foot Wounds

PROVIDER: PRJNA1029592 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26422270.fastq.gz	Fastqsanger.gz
	SRR26422271.fastq.gz	Fastqsanger.gz
	SRR26422272.fastq.gz	Fastqsanger.gz
	SRR26422273.fastq.gz	Fastqsanger.gz
	SRR26422274.fastq.gz	Fastqsanger.gz

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Single Cell Analysis of Gene expression profile as single cell level of chronic diabetic foot ulcers and chronic non-diabetic foot ulcer Foot Wounds

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Gene expression profile as single cell level of chronic diabetic foot ulcers and chronic non-diabetic foot ulcer

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Project description:Diabetic foot ulcers (DFUs) are a devastating complication of diabetes. In order to identify systemic and local factors associated with DFU healing, we examined the cellular landscape of DFUs by single-cell RNA-seq analysis of foot and forearm skin specimens, as well as PBMC samples, from 10 non-diabetic subjects, and 17 diabetic patients, 11 with, and 6 without DFU. Our analysis shows enrichment of a unique inflammatory fibroblast population in DFU patients with healing wounds. The patients with healing DFUs also depicted enrichment of macrophages with M1 polarization, as opposed to more M2 macrophages in non-healing wounds. These findings were verified using Immunohistochemistry and Spatial Transcriptomics.

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Project description:Introduction: We investigated the role and mechanism of hsa-HLA-DRB1 in the development and progression of diabetic foot ulcers. Methods: High-throughput sequencing was performed on three normal foot trauma tissues and diabetic foot ulcer tissues. The circRNAs with significant differences were identified. The downstream miRNAs were predicted by miRanda and RNAhybrid databases, and the mRNAs were predicted by the TargetScan database. Validation was performed with CCK8, flow cytometry, trabecular scratch assay, tubule generation assay, Western blot, dual luciferase assay, and RT-qPCR. Results: High-throughput sequencing identified 461 significantly different circRNAs, of which 260 were up-regulated and 201 down-regulated. Compared to normal tissue, hsa-HLA-DRB1 was highly expressed in diabetic foot ulcers. The hsa-HLA-DRB1/miRNA_12118/FLT-1 axis was constructed. In vitro, we found that HLA-DRB1 overexpression inhibited cell viability, wound healing, and tubule formation, promoted apoptosis, and enhanced FLT-1 expression in HUVECs. Conclusion: The upregulation of hsa-HLA-DRB1 may promote diabetic foot development by targeting miRNA_12118 and acting on FLT-1. Therefore, our study highlights the key role of the hsa-HLA-DRB1/miRNA_12118/FLT-1 axis in diabetic foot trauma.

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