GSE163309
Topical application of an irreversible small molecule inhibitor of Lysyl Oxidases ameliorates skin scarring and fibrosis
Next-generation sequencing (NGS) has been performed to investigate the effect of Lysyl oxidase inhibition by PXS-4787A on transcriptome of normal skin derived fibroblasts and keratinocytes. Fibroblasts and keratinocytes of low passage (p2-5) were cultured onto T25 flasks until ~70% confluence was reached. Fibroblasts were treated with or without 10 µM PXS-4787A in complete fibroblast growth media for 48 hours at 37°C. Keratinocytes were treated with 10 µM PXS-4787A with basal keratinocyte medium supplemented with 1.2 mM calcium chloride for 48 hours at 37°C. Cells were then collected using 0.05% trypsin and washed in Phosphate Buffered Saline (PBS). RNA was extracted using the RNeasy Mini kit protocol. Samples were then sent to the Australian Genome Research Facility (AGRF) where 1µg of RNA was submitted for next-generation sequencing. Once raw data returned, bioinformatics analyses were conducted. This study found that the exposure of PXS-4787A only had minor impact on gene expression in both fibroblasts and keratinocytes. In fibroblasts, 3 genes were significantly upregulated and only one gene was significantly downregulated. Besides, There were only two upregulated genes in the whole genome analysis after LOX inhibition in keratinocytes. In conclusion, this study highlights the safety of LOX-inhibition through PXS-4787A on primary human epidermal cells. The results of the transcriptome analysis intimate that PXS-4787A treatment has no adverse effects on the healthy keratinocytes. Taken together, this study suggests that LOX-inhibition has no potential effects on the epidermal differentiation in human skin. Overall design: 6 normal skin fibroblasts and 5 normal skin keratinocytes with and without treatment of Lysyl Oxidases inhibitor (10 µM PXS-4787A)