OBJECTIVE: This study aimed to investigate new treatment options for AAV through a combination of bioinformatics analysis and cell molecular experiments.
METHODS: The research utilized integrated bioinformatics analysis to identify genes with differential expression, conduct enrichment analysis, and pinpoint hub genes associated with AAV. Potential therapeutic compounds for AAV were identified using Connectivity Map and molecular docking techniques. In vitro experiments were then carried out to examine the impact and mechanism of apilimod on endothelial cell injury induced by MPO-ANCA-positive IgG.
RESULTS: The findings revealed a set of 374 common genes from differentially expressed genes and key modules of WGCNA, which were notably enriched in immune and inflammatory response processes. A proteinprotein interaction network was established, leading to the identification of 10 hub genes, including TYROBP, PTPRC, ITGAM, KIF20A, CD86, CCL20, GAD1, LILRB2, CD8A, and COL5A2. Analysis from Connectivity Map and molecular docking suggested that apilimod could serve as a potential therapeutic cytokine inhibitor for ANCA-GN based on the hub genes. In vitro experiments demonstrated that apilimod could mitigate tight junction disruption, endothelial cell permeability, LDH release, and endothelial activation induced by MPO-ANCA-positive IgG. Additionally, apilimod treatment led to a significant reduction in the expression of proteins involved in the TLR4/NF-κB and NLRP3 inflammasome-mediated pyroptosis pathways.
CONCLUSIONS: This study sheds light on the potential pathogenesis of AAV and highlights the protective role of apilimod in mitigating MPO-ANCA-IgG-induced vascular endothelial cell injury by modulating the TLR4/ NF-kB and NLRP3 inflammasome-mediated pyroptosis pathway. These findings suggest that apilimod may hold promise as a treatment for AAV and warrant further investigation.
目的:本研究旨在通过结合生物信息学分析和细胞分子实验来研究AAV的新治疗方案。
方法:该研究利用整合的生物信息学分析来鉴定具有差异表达的基因,进行富集分析,并精确定位与AAV相关的hub基因。使用ConnectivityMap和分子对接技术鉴定用于AAV的潜在治疗化合物。然后进行体外实验以检查阿吡莫德对MPO-ANCA阳性IgG诱导的内皮细胞损伤的影响和机制。
结果:研究结果揭示了一组374个来自WGCNA差异表达基因和关键模块的常见基因,在免疫和炎症反应过程中尤其丰富。建立了蛋白质相互作用网络,导致10个枢纽基因的鉴定,包括TYROBP,PTPRC,ITGAM,KIF20A,CD86,CCL20,GAD1,LILRB2,CD8A,COL5A2来自ConnectivityMap和分子对接的分析表明,阿吡莫德可以作为基于hub基因的ANCA-GN的潜在治疗性细胞因子抑制剂。体外实验表明,阿吡莫德可以减轻紧密连接的破坏,内皮细胞通透性,LDH释放,MPO-ANCA阳性IgG诱导的内皮激活。此外,阿吡莫德治疗导致参与TLR4/NF-κB和NLRP3炎性体介导的焦亡途径的蛋白质表达显着降低。
结论:本研究揭示了AAV的潜在发病机制,并强调了阿吡莫德通过调节TLR4/NF-kB和NLRP3炎性体介导的细胞凋亡途径减轻MPO-ANCA-IgG诱导的血管内皮细胞损伤的保护作用。这些发现表明,阿吡莫德可能有望作为AAV的治疗方法,并需要进一步调查。