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TRIM32 Inhibits NEK7 Ubiquitylation-Dependent Microglia Pyroptosis After Spinal Cord Injury
Jiasheng Yu1, Dongqian Feng1, Lei Bao1
1Department of Orthopedics, Shuyang Hospital of Traditional Chinese Medicine (Shuyang Hospital of Traditional Chinese Medicine affiliated to Yangzhou University), No. 28, Shanghai Middle Road, Shuyang County, Suqian City, 223600, Jiangsu Province, China.
Abstract:
Spinal cord injury (SCI) is a disabling disease associated with microglial activation. Tripartite motif containing 32 (TRIM32) is an E3 ubiquitin ligase that plays a role in SCI. This study aimed to explore the role of TRIM32 in SCI and its potential mechanisms. We established an SCI mouse model to assess the function of TRIM32 using quantitative real-time polymerase chain reaction (qPCR), and hematoxylin and eosin staining. Additionally, a lipopolysaccharides (LPS)-induced cell injury model was generated to explore the impact of TRIM32 on pyroptosis using qPCR, propidium iodide staining, and western blotting. The ubiquitylation of NEK7 was analyzed using western blotting, co-immunoprecipitation, and immunofluorescence staining. The results showed that TRIM32 expression was increased in SCI mice and LPS-induced BV-2 cells. Overexpression of TRIM32 ameliorated SCI in mice and suppressed pyroptosis in LPS-treated BV-2 cells. Additionally, the E3 ligase TRIM32 promoted the ubiquitylation of NEK7 at the K64 site, leading to the downregulation of NEK7 levels. Inhibiting NEK7 ubiquitylation reversed the suppression of pyroptosis by TRIM32. In conclusion, TRIM32 inhibits microglia pyroptosis by facilitating the ubiquitylation of NEK7 at the K64 site, thereby alleviating the progression of SCI. The findings suggest that TRIM32 has the potential to be a therapeutic target of SCI.
Insights
Tripartite motif containing 32 (TRIM32) reduces spinal cord injury (SCI) by inhibiting microglial pyroptosis. This E3 ubiquitin ligase targets NEK7 for degradation, offering a potential therapeutic strategy for SCI.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Spinal cord injury (SCI) is a debilitating condition linked to microglial activation.
- Tripartite motif containing 32 (TRIM32), an E3 ubiquitin ligase, is implicated in SCI pathogenesis.
Purpose of the Study:
- To investigate the role and underlying mechanisms of TRIM32 in SCI.
- To explore TRIM32's impact on pyroptosis in microglia.
Main Methods:
- Established SCI and lipopolysaccharides (LPS)-induced cell injury mouse models.
- Utilized qPCR, hematoxylin and eosin staining, propidium iodide staining, and western blotting.
- Analyzed NEK7 ubiquitylation via co-immunoprecipitation and immunofluorescence staining.
Main Results:
- TRIM32 expression increased in SCI models and LPS-treated cells.
- TRIM32 overexpression alleviated SCI symptoms and suppressed pyroptosis.
- TRIM32 promoted NEK7 ubiquitylation at K64, downregulating NEK7 levels and inhibiting pyroptosis.
Conclusions:
- TRIM32 inhibits microglia pyroptosis by facilitating NEK7 ubiquitylation at K64, mitigating SCI progression.
- TRIM32 presents a potential therapeutic target for spinal cord injury.
