Zhao Yixuan, Huang Xin, Zhang Zewei, Zhang Yifan, Zhang Guoyou, Zan Tao, Li Qingfeng
Department of Plastic and Reconstructive Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Plastic and Reconstructive Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Front Cell Dev Biol. 2020 Jun 26;8:529. doi: 10.3389/fcell.2020.00529. eCollection 2020.
Re-epithelialization is a fundamental process in wound healing that involves various cytokines and cells during cutaneous barrier reconstruction. Ubiquitin-specific peptidase 15 (), an important member of the deubiquitinating enzymes (DUBs), removes ubiquitin chains from target proteins and maintains protein stability. However, the dynamic role of USP15 in epithelialization remains unclear. We aimed to investigate the regulatory function of USP15 in re-epithelialization. An excisional wound splinting model was established to evaluate the re-epithelialization rate in knockout (KO) mice. Coimmunoprecipitation (Co-IP) and mass spectrum analyses were performed to identify USP15-interacting proteins. RNA-sequencing was performed for transcriptome analysis in keratinocytes and uploaded into NODE database (http://www.biosino.org/node, accession numbers: OEP000770 and OEP000763). First, a significant delay in epithelialization was observed in the KO mice. Moreover, inhibition of cell migration and proliferation was observed in the USP15-silenced keratinocytes (HaCaTs). Moreover, we revealed for the first time that USP15 could interact with eukaryotic initiation factor 4A-1 (EIF4A1), thereby promoting translational efficacy in keratinocytes, which is essential for keratinocyte proliferation and migration. Conclusively, the USP15-EIF4A1 complex significantly accelerated re-epithelialization in wound healing. These observations helped elucidate the function and mechanisms of USP15 in modulating re-epithelialization in wound healing, providing a promising target for refractory wound treatment.
再上皮化是伤口愈合的一个基本过程，在皮肤屏障重建过程中涉及多种细胞因子和细胞。泛素特异性肽酶15（USP15）是去泛素化酶（DUBs）的重要成员，可从靶蛋白上去除泛素链并维持蛋白质稳定性。然而，USP15在上皮化过程中的动态作用仍不清楚。我们旨在研究USP15在再上皮化中的调节功能。建立切除伤口夹板模型以评估基因敲除（KO）小鼠的再上皮化率。进行免疫共沉淀（Co-IP）和质谱分析以鉴定与USP15相互作用的蛋白质。对角质形成细胞进行RNA测序以进行转录组分析，并上传到NODE数据库（http://www.biosino.org/node，登录号：OEP000770和OEP000763）。首先，在USP15基因敲除小鼠中观察到上皮化明显延迟。此外，在USP15沉默的角质形成细胞（HaCaTs）中观察到细胞迁移和增殖受到抑制。此外，我们首次发现USP15可与真核起始因子4A-1（EIF4A1）相互作用，从而促进角质形成细胞中的翻译效率，这对角质形成细胞的增殖和迁移至关重要。总之，USP15-EIF4A1复合物显著加速了伤口愈合中的再上皮化。这些观察结果有助于阐明USP15在调节伤口愈合中再上皮化的功能和机制，为难治性伤口治疗提供了一个有前景的靶点。
