USP13 protects against influenza virus-induced lung injury by promoting IFN-β production

Influenza A virus (IAV) a negative-sense, single-stranded RNA virus, which belongs in the Orthomyxoviridae family. Influenza A is a common cause of acute respiratory illnesses worldwide and associated with considerable morbidity and mortality. Ubiquitin-specific protease 13 (USP13) belongs to the deubiquitinating enzyme (DUB) superfamily. The role of USP13 in the pathogenesis of IAV-induced lung injury has not been revealed. In this study, we investigated the role of USP13 in IAV-induced lung injury. Influenza A/Puerto Rico/8/34 (H1N1) virus was used to infect human lung epithelial cells and mice. IAV infection decreased USP13 protein levels in the mouse lung tissues at 7 days post-infection (dpi). Both USP13 mRNA and protein levels were significantly decreased in IAV-infected A549 cells in dose- and time-dependent manners. Next, we investigated the role of USP13 in IAV-triggered signaling in A549 cells by silencing and overexpressing USP13. Knockdown of USP13 impaired IAV-induced IFN-β mRNA and protein expression, whereas USP13 overexpression enhanced IAV-induced IFN-β mRNA and protein expression. Moreover, knockdown of USP13 significantly increased virus titers at 48 hours post-infection (hpi) in IAV-infected A549 cells. On the other hand, overexpression of USP13 resulted in a significantly decrease in virus titers at 48 hpi. To further investigate the role of USP13 in IAV infection, we infected wild type mice (USP13+/+) and USP13 knockout mice (USP13-/-) with intranasal administration of PR8 virus. USP13-/- mice showed significant more weight loss compared to USP13+/+ mice after PR8 infection. USP13 knockout mice were more susceptible to influenza virus pathology and USP13 deficiency decreases IAV-induced IFN-β production. Mechanistically, Knockdown of USP13 impaired IAV-triggered IRF3 nuclear translocation. Moreover, we found that IAV infection promoted association between USP13 and p-IRF3. Future studies will shed light on the role of USP13 in modulating the ubiquitin-proteasome system in antiviral innate immunity signaling. This work is supported by the National Institutes of Health; R01HL151513 and R01HL167846 to J.Z.; R01AI148180 to Y.A.M; R01AI130110 to J.S.Y. This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.