Kang Sen, Liu Shuxin, Dong Xian, Li Haoyu, Qian Yuanyi, Dai Anna, He Wentao, Li Xiaojun, Chen Qianming, Wang Huiming, Ding Pei-Hui
Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Hangzhou, China.
Autophagy. 2025 Apr;21(4):771-788. doi: 10.1080/15548627.2024.2429371. Epub 2024 Dec 11.
Periodontitis, a prevalent and chronic inflammatory disease, is intricately linked with macroautophagy/autophagy, which has a dual role in maintaining periodontal homeostasis. Despite its importance, the precise interplay between autophagy and periodontitis pathogenesis remains to be fully elucidated. In this study, our investigation revealed that the ubiquitination of RAB7A, mediated by reduced levels of the deubiquitinating enzyme USP4 (ubiquitin specific peptidase 4), disrupts normal lysosomal trafficking and autophagosome-lysosome fusion, thereby contributing significantly to periodontitis progression. Specifically, through genomic and histological analysis of clinical gingival samples, we observed a decreased RAB7A expression and impaired autophagic activity in periodontitis. This was further substantiated through experimental periodontitis mice, where RAB7A inactivation was shown to directly affect autophagy efficiency and drive periodontitis progression. Next, we explored the function of active RAB7A to promote lysosomal trafficking dynamics and autophagosome-lysosome fusion, which was inhibited by RAB7A ubiquitination in macrophages stimulated by (.), one of the keystone pathogens of periodontitis. Last, by proteomics analysis, we revealed that the ubiquitination of RAB7A was mediated by USP4 and validated that upregulation of USP4 could attenuate periodontitis in vivo. In conclusion, these findings highlight the interaction between USP4 and RAB7A as a promising target for therapeutic intervention in managing periodontal diseases. 3-MA: 3-methyladenine; Baf A1:bafilomycin A; BECN1: beclin 1, autophagy related; CEJ-ABC: cementoenamel junctionto alveolar bone crest; IL1B/IL-1β: interleukin 1 beta; KD:knockdown; LPS: lipopolysaccharide; MOI: multiplicity of infection;OE: overexpression; .: ; RILP: Rabinteracting lysosomal protein; ScRNA-seq: single-cell RNA sequencing; SQSTM1/p62: sequestosome 1; .: ; USP4:ubiquitin specific peptidase 4.
牙周炎是一种常见的慢性炎症性疾病，与巨自噬/自噬密切相关，自噬在维持牙周稳态中具有双重作用。尽管其很重要，但自噬与牙周炎发病机制之间的确切相互作用仍有待充分阐明。在本研究中，我们的调查显示，去泛素化酶USP4（泛素特异性肽酶4）水平降低介导的RAB7A泛素化会破坏正常的溶酶体运输和自噬体-溶酶体融合，从而对牙周炎进展有显著影响。具体而言，通过对临床牙龈样本的基因组和组织学分析，我们观察到牙周炎中RAB7A表达降低且自噬活性受损。这在实验性牙周炎小鼠中得到进一步证实，其中RAB7A失活被证明直接影响自噬效率并推动牙周炎进展。接下来，我们探讨了活性RAB7A促进溶酶体运输动态和自噬体-溶酶体融合的功能，在牙周炎的关键病原体之一（.）刺激的巨噬细胞中，RAB7A泛素化抑制了该功能。最后，通过蛋白质组学分析，我们揭示RAB7A的泛素化由USP4介导，并验证了USP4的上调可在体内减轻牙周炎。总之，这些发现突出了USP4与RAB7A之间的相互作用，作为治疗干预牙周疾病的一个有前景的靶点。3-MA：3-甲基腺嘌呤；Baf A1：巴弗洛霉素A；BECN1：贝克林1，自噬相关；CEJ-ABC：牙骨质釉质界至牙槽嵴顶；IL1B/IL-1β：白细胞介素1β；KD：敲低；LPS：脂多糖；MOI：感染复数；OE：过表达；.：；RILP：Rab相互作用溶酶体蛋白；ScRNA-seq：单细胞RNA测序；SQSTM1/p62：聚集体蛋白1；.：；USP4：泛素特异性肽酶4。
