Analysis of skin of keratinocyte-specific OTULIN-deficient mice by single-cell RNA-sequencing

In order to unravel the functional role of the linear deubiquitinase OTULIN, we performed single-cell RNA-sequencing on total skin of mice lacking OTULIN selectively in keratinocytes. Keratinocyte-specific OTULIN knock-out (KO) mice develop delineated inflammatory lesion. Through single-cell analysis on lesional and non-lesional skin of mice lacking OTULIN in keratinocytes, we could identify signalling pathways through which these inflammatory lesions appear, allowing us to get new insights on the molecular events that regulate skin homeostasis and mediate skin inflammation. The purpose of this study was to delineate the cellular and molecular changes that occur in the skin when linear deubiquitination is absent in keratinocytes. Total skin was harvested from wild-type (WT) skin and lesional and non-lesional keratinocyte-specific OTULIN knock-out (KO) mice. We FACS sorted live cells from single-cell suspensions of total skin.为阐明线性去泛素化酶OTULIN的功能角色，我们对角质形成细胞特异性缺失OTULIN的小鼠的全皮肤组织开展了单细胞RNA测序（single-cell RNA-sequencing）。角质形成细胞特异性OTULIN敲除（KO）小鼠可形成边界清晰的炎性皮损。通过对此类角质形成细胞OTULIN缺失小鼠的皮损区与非皮损区皮肤进行单细胞分析，我们成功鉴定出介导此类炎性皮损形成的信号通路，由此为调控皮肤稳态、介导皮肤炎症的分子事件提供了全新的研究见解。本研究旨在阐明当角质形成细胞缺乏线性去泛素化过程时，皮肤内发生的细胞与分子层面的变化。我们从野生型（WT）小鼠的全皮肤，以及角质形成细胞特异性OTULIN敲除（KO）小鼠的皮损区与非皮损区皮肤中采集全皮肤组织，随后通过FACS从全皮肤单细胞悬液中分选活细胞。