Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Department of Dermatology, Brussels, Belgium.
Department of Dermatology and Allergology, Medical School, RWTH Aachen University, Aachen, Germany.
J Eur Acad Dermatol Venereol. 2019 Feb;33(2):367-375. doi: 10.1111/jdv.15301. Epub 2019 Jan 1.
Janus kinase (JAK) inhibition may be a promising new treatment modality for inflammatory (skin) diseases. However, little is known about direct effects of kinase inhibitors on keratinocyte differentiation and function as well as skin barrier formation.
Our aim was to address the direct impact of kinase inhibition of the JAK1/3 pathways by tofacitinib on keratinocyte immune function and barrier formation in atopic dermatitis (AD) and psoriasis.
3D skin equivalents of both diseases were developed and concurrently pretreated with tofacitinib. To induce AD, 3D skin equivalents were stimulated with recombinant human IL-4 and IL-13. Psoriasis-like conditions were induced by incubation with IL-17A, IL-22 and tumour necrosis factor α (TNFα). The activation of signal transducer and activator of transcription (STAT)1, STAT3 and STAT6 was assessed by Western blot analysis. Microarray analysis and quantitative real-time PCR were used for gene expression analysis.
Tofacitinib pretreatment preserved epidermal morphology and reduced STAT3 and STAT6 phosphorylation of AD-like and STAT3 phosphorylation of psoriasis-like culture conditions in 3D skin models compared to sham-controls. Filaggrin expression was fully maintained in the AD-like models, but only partially in psoriasis-like conditions after pretreatment with tofacitinib. In addition, tofacitinib upregulated DSC1, FLG and KRT1. Using gene expression analysis, downregulation of POSTN and IL24 was observed in AD-like conditions, whereas downregulation of IL20 and IL1B was observed in psoriasis-like conditions.
JAK1/3 inhibition counteracted cytokine-induced AD- and psoriasis-like epidermal morphology and enhanced keratinocyte differentiation in 3D skin models. This effect was more pronounced in the AD-like models compared to the psoriasis-like 3D skin models.
Janus 激酶（JAK）抑制可能是一种有前途的治疗炎症（皮肤）疾病的新方法。然而，对于激酶抑制剂对角质形成细胞分化和功能以及皮肤屏障形成的直接影响知之甚少。
我们的目的是研究 JAK1/3 途径的激酶抑制作用对特应性皮炎（AD）和银屑病中角质形成细胞免疫功能和屏障形成的直接影响。
开发了这两种疾病的 3D 皮肤等效物，并同时用托法替尼预处理。为了诱导 AD，3D 皮肤等效物用重组人白细胞介素 4 和白细胞介素 13 刺激。通过孵育白细胞介素 17A、白细胞介素 22 和肿瘤坏死因子 α（TNFα）诱导银屑病样条件。通过 Western blot 分析评估信号转导和转录激活因子（STAT）1、STAT3 和 STAT6 的激活。使用微阵列分析和定量实时 PCR 进行基因表达分析。
与假对照相比，托法替尼预处理保留了 AD 样和银屑病样培养条件下 3D 皮肤模型中的表皮形态，并减少了 STAT3 和 STAT6 的磷酸化。AD 样模型中的丝聚蛋白表达得到充分维持，但在托法替尼预处理后银屑病样条件下仅部分维持。此外，托法替尼上调了 DSC1、FLG 和 KRT1。通过基因表达分析，在 AD 样条件下观察到 POSTN 和 IL24 的下调，而在银屑病样条件下观察到 IL20 和 IL1B 的下调。
JAK1/3 抑制作用可拮抗细胞因子诱导的 AD 和银屑病样表皮形态，并增强 3D 皮肤模型中的角质形成细胞分化。这种作用在 AD 样模型中比在银屑病样 3D 皮肤模型中更为明显。
