YTHDC2-mediated RAB20 degradation regulates NLRP3 inflammasome priming to improve chronic glomerulonephritis

Chronic glomerulonephritis (CGN) primarily arises from immune-mediated inflammation, but the exact targets driving its initiation and progression remain unclear. This study identified the YTHDC2/RAB20/NLRP3 axis as a potential therapeutic target in CGN.

RAB20 and YTHDC2 expression was analyzed through bioinformatics. In vitro, lipopolysaccharide (LPS)-stimulated mouse mesangial cells (MMCs) were transfected with siRNA targeting RAB20 and YTHDC2, along with overexpression plasmids. Actinomycin D was utilized to evaluate RAB20 mRNA stability. RNA immunoprecipitation quantitative PCR (RIP-qPCR) was performed to investigate the interaction between RAB20 mRNA and YTHDC2 protein. Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) and 5‑Ethynyl ‑2'‑Deoxyuridine (EdU) incorporation assays. NLRP3 inflammasome priming was measured by Enzyme-Linked Immunosorbent Assay (ELISA), immunofluorescence, and Western blot (WB). In vivo, adenine-induced CGN mice were treated with adeno-associated virus 9 (AAV9-YTHDC2). Renal function markers were assessed, and kidney histopathology was analyzed via tissue staining.

Both clinical, cellular, and animal models revealed a significant reduction in RAB20 expression in CGN. RAB20 knockdown facilitated NLRP3 inflammasome priming, while its overexpression suppressed these effects. The YTHDC2 protein specifically binds to RAB20 mRNA. YTHDC2 knockdown increased RAB20 mRNA stability, which inhibited MMC proliferation and NLRP3 inflammasome priming. In CGN mice, AAV9-mediated silencing of YTHDC2 improved renal function, as evidenced by reduced kidney function markers and diminished inflammatory cell infiltration.

This study demonstrates that YTHDC2-mediated degradation of RAB20 mRNA regulates NLRP3 inflammasome priming in glomerular mesangial cells and contributes to CGN pathogenesis.