School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK.
Department of Oral Medicine and Oral Surgery, Jordan University of Science and Technology, Irbid, Jordan.
Int Endod J. 2021 Sep;54(9):1571-1580. doi: 10.1111/iej.13547. Epub 2021 Jun 17.
To create an irreversible pulpitis gene signature from microarray data of healthy and inflamed dental pulps, followed by a bioinformatics approach using connectivity mapping to identify therapeutic compounds that could potentially treat pulpitis.
The Gene Expression Omnibus (GEO) database, an international public repository of genomics data sets, was searched for human microarray datasets assessing pulpitis. An irreversible pulpitis gene expression signature was generated by differential expression analysis. The statistically significant connectivity map (ssCMap) method was used to identify compounds with a highly correlating gene expression pattern. qPCR was used to validate novel pulpitis genes. An ex vivo pulpitis model was used to test the effects of the compounds identified, and the level of inflammatory cytokines was measured with qPCR, ELISA and multiplex array. Means were compared using the t-test or ANOVA with the level of significance set at p ≤ .05.
Pulpitis gene signatures were created using differential gene expression analysis at cutoff points p = .0001 and .000018. Top upregulated genes were selected as potential pulpitis biomarkers. Among these, IL8, IL6 and MMP9 were previously identified as pulpitis biomarkers. Novel upregulated genes, chemokine (C-C motif) ligand 21 (CCL21), metallothionein 1H (MT1H) and aquaporin 9 (AQP9) were validated in the pulp tissue of teeth clinically diagnosed with irreversible pulpitis using qPCR. ssCMap analysis identified fluvastatin (Statin) and dequalinium chloride (Quaternary ammonium) as compounds with the strongest correlation to the gene signatures (p = .0001). Fluvastatin reduced IL8, IL6, CCL21, AQP9 (p < .001) and MMP9 (p < .05) in the ex vivo pulpitis model, while dequalinium chloride reduced AQP9 (p < .001) but had no significant effect on the other biomarkers.
AQP9, MT1H and CCL21 were identified and validated as novel biomarkers for pulpitis. Fluvastatin and dequalinium chloride identified by the ssCMap as potential therapeutics for pulpitis reduced selected pulpitis biomarkers in an ex vivo pulpitis model. In vivo testing of these licenced drugs is warranted.
从健康和发炎牙髓的微阵列数据中创建不可逆性牙髓炎基因特征，然后使用连接映射的生物信息学方法来识别可能治疗牙髓炎的治疗化合物。
从评估牙髓炎的人类微阵列数据集的基因表达综合 (GEO) 数据库中搜索国际基因组数据集公共存储库。通过差异表达分析生成不可逆性牙髓炎基因表达特征。使用统计显着性连接映射 (ssCMap) 方法来识别具有高度相关基因表达模式的化合物。使用 qPCR 验证新的牙髓炎基因。使用体外牙髓炎模型测试鉴定出的化合物的作用，并使用 qPCR、ELISA 和多重阵列测量炎症细胞因子的水平。使用 t 检验或方差分析比较均值，显著水平设置为 p≤.05。
使用差异基因表达分析在截止值 p=0.0001 和.000018 处创建牙髓炎基因特征。选择上调的基因作为潜在的牙髓炎生物标志物。其中，IL8、IL6 和 MMP9 先前被鉴定为牙髓炎生物标志物。新型上调基因趋化因子 (C-C 基元) 配体 21 (CCL21)、金属硫蛋白 1H (MT1H) 和水通道蛋白 9 (AQP9) 使用 qPCR 在临床诊断为不可逆性牙髓炎的牙齿牙髓组织中得到验证。ssCMap 分析鉴定出氟伐他汀 (他汀类药物) 和地喹氯铵 (季铵盐) 为与基因特征相关性最强的化合物 (p=0.0001)。氟伐他汀降低了体外牙髓炎模型中的 IL8、IL6、CCL21、AQP9(p<.001) 和 MMP9(p<.05)，而地喹氯铵降低了 AQP9(p<.001)，但对其他生物标志物没有显著影响。
鉴定并验证了 AQP9、MT1H 和 CCL21 作为牙髓炎的新型生物标志物。ssCMap 鉴定的氟伐他汀和地喹氯铵作为牙髓炎的潜在治疗药物，降低了体外牙髓炎模型中选定的牙髓炎生物标志物。需要对这些许可药物进行体内测试。
