整合多组学筛选虎杖治疗干燥综合征的可成药靶点RORA及其作用机制研究

Integrated Multi-Omics Screening of RORA as a Druggable Target of Polygonum cuspidatum for the Treatment of Sjögren’s Syndrome and Investigation of Its Mechanism of Action

    摘要
  • 摘要:
    目的 筛选虎杖治疗干燥综合征（SS）的潜在可成药靶点并研究其作用机制。
    方法 通过网络药理学获取虎杖活性成分及靶点，并与SS疾病靶点、可成药基因集取交集以获得候选靶点，通过孟德尔随机化（SMR）及共定位分析验证因果关联。通过单细胞转录组数据库和免疫组化实验检测候选靶点在SS患者中的表达特征，利用分子对接及100 ns分子动力学模拟评估活性成分与靶点的理论结合活性。
    结果 整合多数据集获取最终核心可成药靶点RORA（OR=0.35，95% CI：0.21~0.58，Pval=4.15E-04，PHEIDI=0.417），共定位分析提示二者由同一遗传变异驱动（PP.H4=0.848）。单细胞转录组显示RORA最主要表达于T细胞、成纤维细胞和B细胞，SS患者唾液腺中RORA总体表达显著降低（P<0.001），但在浸润T细胞中特异性上调。免疫组化结果提示SS患者唇腺组织中RORA蛋白表达显著下调（P<0.001）。分子对接及分子动力学模拟显示β-谷甾醇、木犀草素、鸭脚树叶醛碱与RORA具有较为稳定的动态结合活性，结合自由能分别为-43.91、-27.63、-22.16 kcal·mol-1。KEGG富集分析显示RORA⁺与RORA⁻ T细胞差异表达基因显著富集于抗原加工呈递、Th17细胞分化等免疫相关通路。
    结论 虎杖可能通过多成分靶向RORA调控免疫稳态相关通路治疗SS，RORA可作为SS的潜在可成药靶点。

     

    Abstract:
    OBJECTIVE To screen potential druggable targets of Polygonum cuspidatum for the treatment of Sjögren’s syndrome (SS) and elucidate its underlying molecular mechanism.
    METHODS The active components and corresponding targets of Polygonum cuspidatum were obtained via network pharmacology. Candidate targets were identified by intersecting these targets with SS disease-related targets and the human druggable gene set. The causal association between candidate targets and SS was verified using summary-data-based Mendelian randomization (SMR) and Bayesian colocalization analysis. The expression characteristics of the core target in SS patients were detected by single-cell transcriptomics and immunohistochemistry. The binding activity between active components and the core target was evaluated through molecular docking and 100 ns molecular dynamics simulation.
    RESULTS Integration of multiple datasets identified RORA as the core druggable target (OR=0.35, 95% CI: 0.21-0.58, Pval=4.15E-04, PHEIDI=0.417). Colocalization analysis suggested that both RORA expression and SS susceptibility were driven by the same genetic variant (PP.H4=0.848). Single-cell transcriptomics showed that RORA was predominantly expressed in T cells, followed by fibroblasts and B cells. Its overall expression in the salivary glands of SS patients was significantly decreased (P<0.001), whereas it was specifically upregulated in infiltrating T cells. Immunohistochemistry further verified that RORA protein expression was markedly downregulated in labial gland tissues of SS patients (P<0.001). Molecular docking and molecular dynamics simulation indicated that β-sitosterol, luteolin and Picralinal exhibited stable dynamic binding activity with RORA, with binding free energies of -43.91, -27.63 and -22.16 kcal·mol-1. KEGG enrichment analysis showed that differentially expressed genes between RORA⁺ and RORA⁻ T cells were significantly enriched in immune-related pathways, such as antigen processing and presentation and Th17 cell differentiation.
    CONCLUSION Polygonum cuspidatum exerts therapeutic effects on SS by targeting RORA via multiple active components to regulate immune-inflammatory pathways. RORA may serve as a promising druggable target for SS.

     

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