miRNome and Proteome Profiling of Human Keratinocytes and Adipose Derived Stem Cells Proposed miRNA-Mediated Regulations of Epidermal Growth Factor and Interleukin 1-Alpha

Abstract

Wound healing is regulated by complex crosstalk between keratinocytes and other cell types, including stem cells. In this study, a 7-day direct co-culture model of human keratinocytes and adipose-derived stem cells (ADSCs) was proposed to study the interaction between the two cell types, in order to identify regulators of ADSCs differentiation toward the epidermal lineage. As major mediators of cell communication, miRNome and proteome profiles in cell lysates of cultured human keratinocytes and ADSCs were explored through experimental and computational analyses. GeneChip® miRNA microarray, identified 378 differentially expressed miRNAs; of these, 114 miRNAs were upregulated and 264 miRNAs were downregulated in keratinocytes. According to miRNA target prediction databases and the Expression Atlas database, 109 skin-related genes were obtained. Pathway enrichment analysis revealed 14 pathways including vesicle-mediated transport, signaling by interleukin, and others. Proteome profiling showed a significant upregulation of the epidermal growth factor (EGF) and Interleukin 1-alpha (IL-1α) compared to ADSCs. Integrated analysis through cross-matching the differentially expressed miRNA and proteins suggested two potential pathways for regulations of epidermal differentiation; the first is EGF-based through the downregulation of miR-485-5p and miR-6765-5p and/or the upregulation of miR-4459. The second is mediated by IL-1α overexpression through four isomers of miR-30-5p and miR-181a-5p.

MicroRNA-155 mediates multiple gene regulations pertinent to the role of human adipose-derived mesenchymal stem cells in skin regeneration

“…The next step of the study was to investigate the molecular signalling cascades associated with AD-MSCs' effect in wound healing. This study analysed the previously identified miRNA expression profile in AD-MSCs (Shahin et al, 2023); integrated with the identified proteome signature of AD-MSCs. This integration enabled us to generate a predicted positive regulation Frontiers in Bioengineering and Biotechnology frontiersin.org between miR-155 and genes encoding for FGF2, FGF7, CCL2, and VCAM1 in human AD-MSCs.…”

“…The miRNA and protein screening data in Keratinocytes and AD-MSCs ( Shahin et al, 2023 ), was analysed to identify the upregulated proteins in AD-MSCs. Our reported dataset “Differentially expressed DEmiRNA-mRNA Interaction Network analysis” in AD-MSCs versus keratinocytes was utilized to identify the enriched pathways in AD-MSCs.…”

“…AD-MSCs treated wounds showed better properties of healing that mimicked natural healing. We performed an integrated analysis of our previously published miRNome data in primary human AD-MSCs, as well as to relevant protein content, in comparison to normal keratinocytes as the primary effector in physiological cutaneous wound edge healing ( Shahin et al, 2023 ). The findings were validated through gene expression study and via anti-miR-driven experimental targeting.…”

Identification of biomarkers associated with immune scores in diabetic retinopathy

“…MiR-6752-5p was associated with cerebrovascular disorder ( 69 ). MiR-6858-5p was associated with growth, invasion, and angiogenesis of glioblastoma multiforme ( 70 ), and miR-6765-5p was associated with wound healing ( 71 ). As for miR-6089, it has been reported to have correlation with the risk of ischemic stroke ( 72 ) and rheumatoid arthritis ( 73 ).…”