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LncRNA PVT1 Promotes Intracranial Aneurysm Development via USP10/KLF4/NLRP3 Axis-Mediated Pyroptosis in Human Cerebral Smooth Muscle Cells. | LitMetric
Our previous research identified that lncRNA PVT1 is upregulated in patients with IA. However, the precise functions of PVT1 in IA remain unclear. We compared the levels of PVT1, caspase-3, caspase-1, and NLRP3 in normal and IA patients. The GEO database was utilized to evaluate the expression of KLF4 and NLRP3 in IA. In vitro, we constructed transfected plasmids for silencing (si) and overexpression (oe) of PVT1 and USP10. We assessed cell apoptosis and measured the levels of IL-18, IL-1β, NLRP3, ASC, GSDMD-N, cleaved-caspase-3, and cleaved-caspase-1. CHX chase, immunoprecipitation assays, and bioinformatics tools were employed to evaluate the interactions among PVT1, USP10, and KLF4. Significant differences were observed in the levels of PVT1, KLF4, NLRP3, caspase-3, caspase-1, and histological examinations between normal and IA patients. Compared to the oe-NC group, the levels of IL-18, IL-1β, NLRP3, ASC, GSDMD-N, cleaved-caspase-3, and cleaved-caspase-1 were significantly increased in the oe-PVT1 and oe-USP10 groups. The effect of oe-USP10 was completely inhibited in the oe-USP10+si-PVT1 group. The RIPseq database demonstrated that PVT1 interacts with both USP10 and KLF4. The CHX chase assay showed that KLF4 interacts with both USP10 and PVT1. The RIP assay confirmed the interaction between PVT1 and USP10. The Co-IP assay and UbiBrowser indicated that KLF4 interacts with USP10. The ChIP assay demonstrated that KLF4 interacts with NLRP3. PVT1 may play a role in the pathophysiology of IA by regulating the USP10/KLF4/NLRP3 axis-mediated pyroptosis of HCSMCs.
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