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Human Single Cell Transcriptional Profiling Identifies Epigenetically Altered PGE2 Pathway that Modulates Diabetic Macrophage Phenotype and Wound Repair

ABSTRACT: Human Single Cell Transcriptional Profiling Identifies Epigenetically Altered PGE2 Pathway that Modulates Diabetic Macrophage Phenotype and Wound Repair

PROVIDER: PRJNA646693 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12235469_1.fastq.gz	Fastqsanger.gz
	SRR12235469_2.fastq.gz	Fastqsanger.gz
	SRR12235470_1.fastq.gz	Fastqsanger.gz
	SRR12235470_2.fastq.gz	Fastqsanger.gz
	SRR12235471_1.fastq.gz	Fastqsanger.gz

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Human Single Cell Transcriptional Profiling Identifies Epigenetically Altered PGE2 Pathway that Modulates Diabetic Macrophage Phenotype and Wound Repair

Project description:Human Single Cell Transcriptional Profiling Identifies Epigenetically Altered PGE2 Pathway that Modulates Diabetic Macrophage Phenotype and Wound Repair

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Human Single Cell Transcriptional Profiling Identifies Epigenetically Altered PGE2 Pathway that Modulates Diabetic Macrophage Phenotype and Wound Repair

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SFRP2 modulates functional phenotype transition and energy metabolism of macrophages during diabetic wound healing [Skin wound]

Project description:Diabetic foot ulcer (DFU) is a serious complication of diabetes mellitus, which causes great health damage and economic burden to patients. The pathogenesis of DFU is not fully understood.We screened wound healing-related genes using bioinformatics analysis, and full-thickness skin injury mice model and cellular assays were used to explore the role of target genes in diabetic wound healing. SFRP2 was identified as a wound healing-related gene, and the expression of SFRP2 is associated with immune cell infiltration in DFU. In vivo study showed that suppression of SFRP2 delayed the wound healing process of diabetic mice, impeded angiogenesis and matrix remodeling, and increased macrophage infiltration in wound tissues. In addition, suppression of SFRP2 enhanced M1 polarization in both the early and later stage of wound healing, and decreased M2 polarization in the later stage, which impeded the transition of M1 to M2 polarization of wound healing. Moreover, suppression of SFRP2 affected the transcriptome signatures-related to inflammatory response and energy metabolism at the early stage of wound healing. Extracellular flux analysis (EFA) showed that suppression of SFRP2 decreased mitochondrial energy metabolism and increased glycolysis in injury-related macrophages. Furthermore, suppression of SFRP2 inhibited transcriptome signaturesrelated to carbohydrate metabolism, lipid metabolism and amino acid metabolism, which consists the three main components of energy metabolism of macrophages. In conclusions, SFRP2 may function as a wound healing-related gene in DFU, and suppression of SFRP2 impaired diabetic wound healing by compromising the M1-to-M2 transition of macrophages and modulating the balance between mitochondrial energy metabolism and glycolysis.

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Differentially Expressed Wound Healing-Related microRNAs in the Human Diabetic Cornea

Project description:MicroRNAs are powerful gene expression regulators, but their corneal repertoire and potential changes in corneal diseases remain unknown. Our purpose was to identify miRNAs altered in the human diabetic cornea by microarray analysis, and to examine their effects on wound healing in cultured telomerase-immortalized human corneal epithelial cells (HCEC) in vitro. Using microarrays, 29 miRNAs were identified as differentially expressed in diabetic samples. Two miRNA candidates showing the highest fold increased in expression in the diabetic cornea were confirmed by Q-PCR and further characterized. HCEC transfection with h-miR-146a or h-miR-424 significantly retarded wound closure, but their respective antagomirs significantly enhanced wound healing vs. controls. Cells treated with h-miR-146a or h-miR-424 had decreased p-p38 and p-EGFR staining, but these increased over control levels close to the wound edge upon antagomir treatment. In conclusion, several miRNAs with increased expression in human diabetic central corneas were found. Two such miRNAs inhibited cultured corneal epithelial cell wound healing. Dysregulation of miRNA expression in human diabetic cornea may be an important mediator of abnormal wound healing. Total RNA was extracted from age-matched human autopsy normal (n=6) and diabetic (n=6) central corneas, Flash Tag end-labeled, and hybridized to Affymetrix® GeneChip® miRNA Arrays. Select miRNAs associated with diabetic cornea were validated by quantitative RT-PCR (Q-PCR) and by in situ hybridization (ISH) in independent samples.

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