Biological Science Research, Kao Corporation, Tochigi, Japan.
Allergy Center, National Center for Child Health and Development, Tokyo, Japan.
J Eur Acad Dermatol Venereol. 2022 Sep;36(9):1477-1485. doi: 10.1111/jdv.18173. Epub 2022 May 13.
Specimens for analysing the molecular pathology of skin disease are generally obtained through invasive methods, such as biopsy. However, less burdensome methods are desirable for paediatric patients. We recently established a method that comprehensively analyses RNA present in sebum (skin surface lipid-RNAs: SSL-RNAs) using a next-generation sequencer. Using this method, biological information can be obtained from the skin in a completely non-invasive manner.
To verify the applicability of the SSL-RNA method for analysis of paediatric skin and analyse the molecular pathology of mild-to-moderate atopic dermatitis (AD) in children.
We collected sebum specimens from the whole faces of 23 healthy children and 16 children with mild-to-moderate AD (eczema area and severity index (EASI) score: 5.9 ± 2.6) ranging in age from 6 months to 5 years, using an oil-blotting film. We then extracted SSL-RNAs from the samples and performed an AmpliSeq transcriptomic analysis.
The expressions of genes related to keratinization (LCE, PSORS1C2, IVL and KRT17), triglyceride synthesis and storage (PLIN2, DGAT2 and CIDEA), wax synthesis (FAR2), ceramide synthesis (GBA2, SMPD3 and SPTLC3), antimicrobial peptides (DEFB1) and intercellular adhesion (CDSN), all of which are related to the skin barrier, are lower in children with AD than in healthy children. The children with AD also have higher expression of CCL17, a Th2-cytokine and an increased Th2-immune response as demonstrated by a gene set variation analysis. Moreover, KRT17 and CCL17 expression levels are significantly correlated with the EASI score.
Molecular changes associated with abnormal immune responses and the epidermal barrier in children with mild-to-moderate AD can be determined using the SSL-RNA method. This non-invasive method could therefore be a useful means for understanding the molecular pathology of paediatric AD.
用于分析皮肤疾病分子病理学的标本通常通过活检等有创方法获得。然而，对于儿科患者来说，希望采用负担更小的方法。我们最近建立了一种使用下一代测序仪全面分析皮脂中 RNA（皮肤表面脂质 RNA：SSL-RNAs）的方法。使用这种方法，可以以完全非侵入性的方式从皮肤中获取生物学信息。
验证 SSL-RNA 方法在分析儿科皮肤中的适用性，并分析儿童轻度至中度特应性皮炎（AD）的分子病理学。
我们使用吸油膜片收集了 23 名健康儿童和 16 名年龄在 6 个月至 5 岁之间患有轻度至中度 AD（湿疹面积和严重程度指数（EASI）评分：5.9±2.6）儿童的整个面部的皮脂标本。然后，我们从这些样本中提取 SSL-RNAs，并进行 AmpliSeq 转录组分析。
AD 患儿角质化相关基因（LCE、PSORS1C2、IVL 和 KRT17）、甘油三酯合成和储存（PLIN2、DGAT2 和 CIDEA）、蜡合成（FAR2）、神经酰胺合成（GBA2、SMPD3 和 SPTLC3）、抗菌肽（DEFB1）和细胞间黏附（CDSN）的表达均低于健康儿童，这些基因均与皮肤屏障有关。AD 患儿还表现出更高的 Th2 细胞因子 CCL17 的表达，以及通过基因集变异分析显示的 Th2 免疫反应增强。此外，KRT17 和 CCL17 的表达水平与 EASI 评分显著相关。
使用 SSL-RNA 方法可以确定患有轻度至中度 AD 的儿童与异常免疫反应和表皮屏障相关的分子变化。因此，这种非侵入性方法可能是了解儿科 AD 分子病理学的有用手段。
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