METHODS: GEO datasets were analyzed to investigate the association of TOPK with psoriasis. Skin immunohistochemical (IHC) staining was performed to clarify the major cells expressing TOPK. TOPK conditional knockout (cko) mice were used to investigate the role of TOPK-specific deletion in IMQ-induced psoriasis-like dermatitis in mice. Flow cytometry was used to analyze the alteration of psoriasis-related immune cells in the lesional skin. Next, the M5-induced psoriasis cell model was used to identify the potential mechanism by RNA-seq, RT-RCR, and western blotting. Finally, the neutrophil-neutralizing antibody was used to confirm the relationship between TOPK and neutrophils in psoriasis-like dermatitis in mice.
RESULTS: We found that TOPK levels were strongly associated with the progression of psoriasis. TOPK was predominantly increased in the epidermal keratinocytes of psoriatic lesions, and conditional knockout of TOPK in keratinocytes suppressed neutrophils infiltration and attenuated psoriatic inflammation. Neutrophils deletion by neutralizing antibody greatly diminished the suppressive effect of TOPK cko in psoriasis-like dermatitis in mice. In addition, topical application of TOPK inhibitor OTS514 effectively attenuated already-established psoriasis-like dermatitis in mice. Mechanismly, RNA-seq revealed that TOPK regulated the expression of some genes in the IL-17 signaling pathway, such as neutrophils chemokines CXCL1, CXCL2, and CXCL8. TOPK modulated the expression of neutrophils chemokines via activating transcription factors STAT3 and NF-κB p65 in keratinocytes, thereby promoting neutrophils infiltration and psoriasis progression.
CONCLUSIONS: This study identified a crucial role of TOPK in psoriasis by regulating neutrophils infiltration, providing new insights into the pathogenesis of psoriasis.
方法:分析GEO数据集,以研究TOPK与银屑病的相关性。进行皮肤免疫组织化学(IHC)染色以澄清表达TOPK的主要细胞。TOPK条件敲除(cko)小鼠用于研究TOPK特异性缺失在IMQ诱导的小鼠牛皮癣样皮炎中的作用。流式细胞术用于分析皮损中银屑病相关免疫细胞的变化。接下来,M5诱导的银屑病细胞模型用于通过RNA-seq鉴定潜在机制,RT-RCR,和西方印迹。最后,中性粒细胞中和抗体用于确认小鼠银屑病样皮炎中TOPK与中性粒细胞之间的关系.
结果:我们发现TOPK水平与银屑病的进展密切相关。TOPK主要在银屑病皮损的表皮角质形成细胞中增加,和条件敲除角质形成细胞中的TOPK抑制中性粒细胞浸润和减轻银屑病炎症。通过中和抗体消除中性粒细胞大大降低了TOPKcko对小鼠牛皮癣样皮炎的抑制作用。此外,局部应用TOPK抑制剂OTS514可有效减轻小鼠中已经确定的银屑病样皮炎.机械地,RNA-seq显示TOPK调控IL-17信号通路中一些基因的表达,例如中性粒细胞趋化因子CXCL1、CXCL2和CXCL8。TOPK通过激活转录因子STAT3和NF-κBp65调节中性粒细胞趋化因子在角质形成细胞中的表达,从而促进中性粒细胞浸润和银屑病进展。
结论:本研究通过调节中性粒细胞浸润确定了TOPK在银屑病中的关键作用,为银屑病的发病机制提供新的见解。