Next Generation Sequencing of skin from wild-type and ADAM17 cKO (Sox9-Cre x ADAM17fl/fl) mice

Purpose: Transcriptomic analysis of ADAM17 cKO mice, a mouse model of atopic dermatitis. Methods: Skin was harvested from 10-week-old wild-type and ADAM17 cKO mice and were subjected to next generation sequencing as described. Results: Of 13,573 genes analyzed by GSA, 2514 differentially expressed genes (DEGs) were detected in ADAM17 cKO skin using a standard definition including both statistically significant change (P-value < 0.01; FDR < 0.05) and effect size (|fold change| > 2) . These DEGs included genes important in lipid metabolism and keratinization, consistent with the understanding of barrier dysfunction in atopic inflammation, as well as significant immunologic pathways in adaptive and innate immunity previously identified in the human AD transcriptome Conclusions: Our study reveals that the transcriptome of ADAM17 cKO mice capture a part of upregulated genes previously observed in human AD skin. Epidermal and whole skin mRNA profiles of 10-week-old WT and ADAM17 cKO mice were generated by deep sequencing, in triplicates.研究目的：针对特应性皮炎小鼠模型ADAM17条件性敲除（cKO，conditional knockout）小鼠开展转录组分析。 实验方法：采集10周龄野生型（wild-type, WT）与ADAM17 cKO小鼠的皮肤组织，按照既定方案开展下一代测序（next generation sequencing, NGS）。 实验结果：通过基因集分析（Gene Set Analysis, GSA）对13573个基因进行分析后，采用同时包含统计学显著性差异（P值<0.01；错误发现率（False Discovery Rate, FDR）<0.05）与效应量（|折叠变化|>2）的标准判定规则，在ADAM17 cKO小鼠皮肤样本中检测到2514个差异表达基因（differentially expressed genes, DEGs）。上述DEGs涵盖脂质代谢与角质形成相关的关键基因，与特应性炎症中皮肤屏障功能障碍的现有认知相符，同时也涉及既往在人类特应性皮炎（atopic dermatitis, AD）转录组中鉴定出的适应性免疫与固有免疫相关的显著免疫通路。 研究结论：本研究证实，ADAM17 cKO小鼠的转录组可捕捉到既往在人类AD皮肤中观察到的部分上调基因。本研究通过深度测序（deep sequencing）获得了10周龄WT与ADAM17 cKO小鼠的表皮及全皮肤mRNA表达谱，实验设置三次生物学重复。