Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations.
Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations.
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DOI：
10.3390/genes13101874
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发表时间：
2022-10-16
期刊：
影响因子：
3.5
通讯作者：
Hicks, Steven D.
中科院分区：
文献类型：
--
作者：
Sullivan, Rhea;Montgomery, Austin;Scipioni, Anna;Jhaveri, Pooja;Schmidt, Adam T.;Hicks, Steven D.
唾液microRNA（miRNAs）作为人类疾病的非侵入性生物标志物越来越受到关注。这种方法需要了解实验设计的差异如何影响miRNA表达。技术方法的变化，加上个体间的差异可能会降低研究的重现性和普遍性。唾液miRNA生物标志物研究面临的另一个障碍是缺乏公认的“对照miRNA”。在迄今为止最大规模的人类唾液miRNA研究之一（922名健康个体）中，我们利用1225份唾液样本来量化由比对器选择（Bowtie1与Bowtie2）、唾液收集方法（痰与拭子）、RNA稳定剂（存在与不存在）和个体生物因素（性别、年龄、体重指数、运动、热量摄入）引起的miRNA表达的变异性。差异表达分析显示，RNA稳定剂的情况下引入了最大的变异性，其次是收集和比对方法的差异。生物学因素通常影响较少数量的miRNAs。我们还报告了643个miRNAs在唾液中的变异系数，突出了几个唾液miRNAs作为参考基因。因此，该分析的结果可以被研究人员用于优化唾液miRNA测量的参数，排除被许多生物因素混淆的miRNA，并确定适当的miRNA对照。
There is growing interest in saliva microRNAs (miRNAs) as non-invasive biomarkers for human disease. Such an approach requires understanding how differences in experimental design affect miRNA expression. Variations in technical methodologies, coupled with inter-individual variability may reduce study reproducibility and generalizability. Another barrier facing salivary miRNA biomarker research is a lack of recognized “control miRNAs”. In one of the largest studies of human salivary miRNA to date (922 healthy individuals), we utilized 1225 saliva samples to quantify variability in miRNA expression resulting from aligner selection (Bowtie1 vs. Bowtie2), saliva collection method (expectorated vs. swabbed), RNA stabilizer (presence vs. absence), and individual biological factors (sex, age, body mass index, exercise, caloric intake). Differential expression analyses revealed that absence of RNA stabilizer introduced the greatest variability, followed by differences in methods of collection and aligner. Biological factors generally affected a smaller number of miRNAs. We also reported coefficients of variations for 643 miRNAs consistently present in saliva, highlighting several salivary miRNAs to serve as reference genes. Thus, the results of this analysis can be used by researchers to optimize parameters of salivary miRNA measurement, exclude miRNAs confounded by numerous biologic factors, and identify appropriate miRNA controls.
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