MF-094, a potent and selective USP30 inhibitor, accelerates diabetic wound healing by inhibiting the NLRP3 inflammasome

• Published in: Experimental cell research 2022-01, Vol.410 (2), p.112967, Article 112967
• Main Authors: Li, Xu, Wang, Tao, Tao, Yue, Wang, Xiaojun, Li, Limeng, Liu, Jianjun
• Format: Article
• Language: English
• Subjects: Animals; Cell Line; Cell Movement - drug effects; Cell Survival - drug effects; Diabetes Mellitus - pathology; Diabetic Foot - pathology; Diabetic foot ulcers; Disease Models, Animal; Down-Regulation - drug effects; Female; Glycation End Products, Advanced - metabolism; Humans; Inflammasome; Inflammasomes - metabolism; Male; Middle Aged; Mitochondrial Proteins - antagonists & inhibitors; Mitochondrial Proteins - metabolism; Naphthalenes - pharmacology; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; NLRP3; Rats; Rats, Sprague-Dawley; Thiolester Hydrolases - antagonists & inhibitors; Thiolester Hydrolases - metabolism; Ubiquitination - drug effects; USP30; Wound healing; Wound Healing - drug effects
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2021.112967

Diabetes is a prevalent disease worldwide that can result in several complications, including renal failure, blindness, and amputation. Diabetic foot ulcers, which have the characteristics of chronic wounds, are a devastating component of diabetes progression that can lead to lower extremity amputation. In this study, we set out to investigate the mechanisms involved in wound healing of diabetic foot ulcers. The expression of USP30 in skin tissues of patients with diabetic foot ulcers and HSF2 human skin fibroblasts treated with advanced glycation end (AGE) products was detected by qRT-PCR, and CCK-8, cell scratch and ELISA assay were used to detect cell viability, migration and levels of Col I, Col III, MMP-2, MMP-9, IL-1β and IL-18. The interaction between USP30 and NLRP3 was verified by co-immunoprecipitation and ubiquitination assays. The expression of USP30, NLRP3 and caspase-1 p20 was detected by Western blot. USP30 inhibitor MF-094 was used to treat diabetic rat model established by streptozotocin (STZ). We found that USP30, a deubiquitinase, was upregulated in skin tissues of patients with diabetic foot ulcers compared with normal skin tissues. In vitro, we found that treatment of HSF2 human skin fibroblasts with advanced glycation end (AGE) products, known to contribute to diabetic complications, resulted in suppressed viability and migration of HSF2 cells, as well as increased levels of USP30 mRNA and protein. Functionally, downregulation of USP30 via shRNA-mediated knockdown or treatment with the USP30 inhibitor MF-094, restored viability and migration of AGE-treated HSF2 cells. We identified the NLRP3 inflammasome as a critical target of USP30 in AGE-induced functions. Mechanistically, we demonstrate that USP30 activates the NLRP3 inflammasome by deubiquitinating NLRP3. Finally, we show that inhibition of USP30 via MF-094 treatment facilitated wound healing in diabetic rats and resulted in decreased protein levels of NLRP3 and its downstream target caspase-1 p20, thus establishing the physiological importance of the identified USP30-NLRP3 link. Together, our findings suggest a therapeutic potential for USP30 in diabetic foot ulcers.