USP16 drives psoriasis progression by deubiquitinating and stabilizing NLRP3 in keratinocytes
Nan Wang, Fangqian Guan, Yifan Lin, Bohao Sun, Jindan Dai, Xiejun Xu, Weibo Tang, Yanhua Ren, Xuliang Huang, Wenjie Gao, Xixi Chen, Litai Jin, Weitao Cong, Zhongxin Zhu
Nan Wang, Fangqian Guan, Yifan Lin, Bohao Sun, Jindan Dai, Xiejun Xu, Weibo Tang, Yanhua Ren, Xuliang Huang, Wenjie Gao, Xixi Chen, Litai Jin, Weitao Cong, Zhongxin Zhu
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Research Article Cell biology Dermatology

USP16 drives psoriasis progression by deubiquitinating and stabilizing NLRP3 in keratinocytes

Abstract

Psoriasis is a chronic inflammatory dermatosis characterized by pathological keratinocyte hyperproliferation and dysregulated immune activation. While ubiquitin-specific peptidase 16 (USP16) has been implicated in modulating multiple cellular signaling pathways, its functional role in psoriatic pathogenesis remains poorly understood. Our investigation revealed pronounced upregulation of USP16 expression in psoriatic epidermis compared with normal controls. Keratinocyte-specific USP16 knockdown demonstrated remarkable therapeutic efficacy, significantly ameliorating characteristic psoriatic phenotypes including epidermal hyperplasia and inflammatory infiltration. RNA-seq analysis showed that USP16 has substantial effects on cell cycle transition and keratinocytes proliferation. Through KEGG analysis, it was found that USP16 primarily regulates the NLRP3 signaling pathway, leading to enhanced cell proliferation and inflammation. Mechanically, USP16 directly binds to the NLRP3 protein to eliminate K48 ubiquitination modification, enhancing the stability of the NLRP3 protein, activating inflammasome activity. Further studies showed that the therapeutic effects of reducing USP16 on psoriasis progression were counteracted by an NLRP3 activator and keratinocyte-specific NLRP3 overexpression adenovirus. Collectively, these results shed light on how USP16 promotes NLRP3 signaling in keratinocytes, exacerbating psoriasis development. This positive regulation highlights the potential of USP16 as a therapeutic target for psoriasis.

Authors

Nan Wang, Fangqian Guan, Yifan Lin, Bohao Sun, Jindan Dai, Xiejun Xu, Weibo Tang, Yanhua Ren, Xuliang Huang, Wenjie Gao, Xixi Chen, Litai Jin, Weitao Cong, Zhongxin Zhu

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Figure 3

USP16 is required for keratinocyte proliferation.

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USP16 is required for keratinocyte proliferation. (A) RNA-seq analysis o...
(A) RNA-seq analysis of the differentially regulated genes between si-Scr group and si-USP16 group treated with M5 for 24 hours. Differentially regulated genes were enriched in GO biological process terms. (B) GSEA results for cell cycle transition. (C) Immunoblotting and quantitative analysis of USP16, Cyclin A1, Cyclin E1, and Bcl-2 protein levels in HaCaT cells that were treated with si-Scr or si-USP16 and stimulated with or without M5 for 24 hours. β-Actin was used as a loading control (n = 3). (D) Immunofluorescence and quantitative analysis of Ki-67+ in HaCaT cells that were treated with si-Scr or si-USP16 and stimulated with or without M5 for 24 hours. Nuclei were stained with DAPI (blue) (n = 3). Scale bar: 50 μm. (E) Immunofluorescent and quantitative analysis of EdU+ in HaCaT cells that were treated with si-Scr or si-USP16 and stimulated with or without M5 for 24 hours. Nuclei were stained with Hoechst (blue) (n = 3). Scale bar: 50 μm. (F) Flow cytometric plots of cell-cycle analysis performed with PI staining on HaCaT cells treated with si-Scr or si-USP16 and stimulated with or without M5 for 24 hours (left). Quantification the percentage of cells that fall into the sub G0/G1, S, or G2/M gates (right). Data are shown as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. The P value was determined using 1-way ANOVA (C–F). All numbers (n) are biologically independent experiments.