Kahraman Mustafa, Laufer Thomas, Backes Christina, Schrörs Hannah, Fehlmann Tobias, Ludwig Nicole, Kohlhaas Jochen, Meese Eckart, Wehler Thomas, Bals Robert, Keller Andreas
Clinical Bioinformatics, Saarland University, Homburg, Germany.
Hummingbird Diagnostics GmbH, Heidelberg, Germany.
Clin Chem. 2017 Sep;63(9):1476-1488. doi: 10.1373/clinchem.2017.271619. Epub 2017 Jul 5.
Different work flows have been proposed to use miRNAs as blood-borne biomarkers. In particular, the method used for collecting blood from patients can considerably influence the diagnostic results.
We explored whether dried blood spots (DBSs) facilitate stable miRNA measurements and compared its technical stability with biological variability. First, we tested the stability of DBS samples by generating from 1 person 18 whole-genome-wide miRNA profiles of DBS samples that were exposed to different temperature and humidity conditions. Second, we investigated technical reproducibility by performing 7 replicates of DBS again from 1 person. Third, we investigated DBS samples from 53 patients with lung cancer undergoing different therapies. Across these 3 stages, 108 genome-wide miRNA profiles from DBS were generated and evaluated biostatistically.
In the stability analysis, we observed that temperature and humidity had an overall limited influence on the miRNomes (average correlation between the different conditions of 0.993). Usage of a silica gel slightly diminished DBS' technical reproducibility. The 7 technical replicates had an average correlation of 0.996. The correlation with whole-blood PAXGene miRNomes of the same individual was remarkable (correlation of 0.88). Finally, evaluation of the samples from the 53 patients with lung cancer exposed to different therapies showed that the biological variations exceeded the technical variability significantly ( < 0.0001), yielding 51 dysregulated miRNAs.
We present a stable work flow for profiling of whole miRNomes on the basis of samples collected from DBS. Biological variations exceeded technical variations significantly. DBS-based miRNA profiles will potentially further the translational character of miRNA biomarker studies.
已经提出了不同的工作流程来将微小RNA（miRNA）用作血液中的生物标志物。特别是，从患者采集血液的方法会对诊断结果产生很大影响。
我们探讨了干血斑（DBS）是否有助于稳定地测量miRNA，并将其技术稳定性与生物学变异性进行了比较。首先，我们通过从1个人生成18个全基因组范围的DBS样本miRNA谱来测试DBS样本的稳定性，这些样本暴露于不同的温度和湿度条件下。其次，我们通过再次从1个人进行7次DBS重复实验来研究技术可重复性。第三，我们研究了53例接受不同治疗的肺癌患者的DBS样本。在这3个阶段中，共生成了108个来自DBS的全基因组范围的miRNA谱，并进行了生物统计学评估。
在稳定性分析中，我们观察到温度和湿度对miRNA组的总体影响有限（不同条件之间的平均相关性为0.993）。使用硅胶会稍微降低DBS的技术可重复性。7次技术重复的平均相关性为0.996。与同一个体的全血PAXGene miRNA组的相关性显著（相关性为0.88）。最后，对53例接受不同治疗的肺癌患者的样本进行评估，结果显示生物学变异显著超过技术变异（<0.0001），产生了51个失调的miRNA。
我们提出了一种基于DBS采集样本进行全miRNA组分析的稳定工作流程。生物学变异显著超过技术变异。基于DBS的miRNA谱可能会进一步推动miRNA生物标志物研究的转化特性。
