Data_Sheet_1_Profiling of Differentially Expressed MicroRNAs in Saliva of Parkinson's Disease Patients.docx

Objective: This study aims to identify differentially expressed salivary miRNAs and validate the diagnostic potential for idiopathic Parkinson's disease (PD). Also, the disease specificity of candidate miRNAs was evaluated between PD, multiple system atrophy (MSA), and essential tremor (ET).Methods: We collected salivary samples from 50 PD, 20 ET, and 20 MSA patients, as well as 30 healthy controls (HCs). In the discovery phase, salivary miRNA microarray analysis was performed. In-silico analysis was used to investigate the target genes of differentially expressed miRNAs and clustered pathways. In validation phase, RT-qPCR was performed with samples from 30 PD patients and 30 HCs. Subsequently, we investigated candidate miRNAs in all recruited subjects. Receiver operating characteristic curve and Spearman correlation analysis was performed to determine diagnostic usefulness.Results: We identified 43 miRNAs that were differentially expressed between 5 PD patients and 5 HCs by miRNA microarray analysis. Computational analysis revealed the target genes were clustered in the pathways associated with ubiquitin protein ligase activity. The result of RT-qPCR showed that the miR-29a-3p and miR-29c-3p were found to be significantly downregulated (p = 0.004, p = 0.027), whereas the miR-6756-5p was significantly upregulated in 30 PD patients compared with 30 HCs (p = 0.032). The miR-29a-3p expression level in PD patients was significantly lower than ET patients (p = 0.035), but higher than MSA patients (p < 0.0001). The diagnostic efficacy reached a little higher when the combination of miR-29a-3p and miR-29c-3p.Conclusion: The miRNA combination of salivary miR-29a-3p and miR-29c-3p has potential to be a diagnostic biomarker for idiopathic PD.

研究目的：本研究的目的是鉴定差异表达的唾液miRNA，并验证其作为原发性帕金森病（PD）诊断潜力的有效性。同时，评估候选miRNA在PD、多系统萎缩（MSA）和特发性震颤（ET）之间的疾病特异性。研究方法：我们收集了50例PD患者、20例ET患者和20例MSA患者的唾液样本，以及30名健康对照者（HCs）的唾液样本。在发现阶段，进行了唾液miRNA微阵列分析。通过计算机分析研究差异表达miRNA的目标基因和聚类通路。在验证阶段，对30例PD患者和30名HCs的样本进行了实时定量PCR分析。随后，我们对所有招募的受试者的候选miRNA进行了研究。通过受试者工作特征曲线和Spearman相关分析确定诊断价值。研究结果：通过miRNA微阵列分析，我们鉴定出43种在5例PD患者和5名HCs之间存在差异表达的miRNA。计算分析显示，目标基因聚集在涉及泛素蛋白连接酶活性的通路中。实时定量PCR的结果表明，miR-29a-3p和miR-29c-3p的表达水平显著下调（p = 0.004，p = 0.027），而miR-6756-5p在30例PD患者中的表达水平与30名HCs相比显著上调（p = 0.032）。PD患者中的miR-29a-3p表达水平显著低于ET患者（p = 0.035），但高于MSA患者（p < 0.0001）。当结合miR-29a-3p和miR-29c-3p时，诊断效能略有提升。研究结论：唾液miR-29a-3p和miR-29c-3p的组合具有作为原发性PD诊断生物标志物的潜力。