Department of Pediatric Research, Oslo University Hospital, Rikshospitalet, Oslo, Norway.
Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
Pediatr Res. 2021 May;89(7):1780-1787. doi: 10.1038/s41390-020-01153-3. Epub 2020 Sep 15.
Circulatory miRNAs are promising biomarkers. The feasibility of using miRNA from dried blood spots (DBS) was investigated using newborn screening cards from patients with cholestasis-lymphedema syndrome (Aagenaes syndrome) and controls.
Total amount of miRNA and specific miRNAs from DBS were analyzed. miRNA was also obtained from newborn screening cards in patients with cholestasis-lymphedema syndrome/Aagenaes syndrome and in healthy newborns.
No differences in miRNA concentrations were found between multispotted samples and samples with one single drop of blood and between central and peripheral punches. Ten repeated freeze-thaw cycles did not significantly change miRNA levels from controls. miR-299 (1.73-fold change, p = 0.034) and miR-365 (1.46-fold change, p = 0.011) were upregulated and miR-30c (0.72-fold change, p = 0.0037), miR-652 (0.85-fold change, p = 0.025), and miR-744 (0.72-fold change, p = 0.0069) were downregulated in patients with Aagenaes syndrome at birth compared to controls.
miRNAs were not affected by multispotting or punch location and were stable throughout repeated freeze-thaw cycles. miRNA in dried blood spots could be used to detect differential expression of miRNA in newborns with Aagenaes syndrome and healthy controls in newborn screening cards. Dried blood spots may be a useful source to explore circulating miRNA as biomarkers.
Circulating miRNAs can be useful biomarkers. miRNAs from dried blood spots were not affected by multispotting or punch location and were stable throughout repeated freeze-thaw cycles. Discrimination between patients and controls are allowed even with few individuals. Early after birth, patients with cholestasis-lymphedema syndrome exhibit miRNA profiles associated with liver fibrosis. This study demonstrated that newborn screening cards may be a useful source for studying miRNA as the technical variability is smaller than biological variation.
循环 miRNA 是很有前途的生物标志物。本研究使用胆汁淤积性淋巴水肿综合征（Aagenaes 综合征）患者和对照者的新生儿筛查卡，考察了从干血斑（DBS）中提取 miRNA 的可行性。
分析 DBS 中的 miRNA 总量和特定 miRNA。还从胆汁淤积性淋巴水肿综合征/Aagenaes 综合征患者和健康新生儿的新生儿筛查卡中提取 miRNA。
多斑点样本与单滴血样本以及中央和外周采血点样本之间的 miRNA 浓度无差异。10 次冻融循环不会显著改变对照者的 miRNA 水平。miR-299（上调 1.73 倍，p=0.034）和 miR-365（上调 1.46 倍，p=0.011）上调，miR-30c（下调 0.72 倍，p=0.0037）、miR-652（下调 0.85 倍，p=0.025）和 miR-744（下调 0.72 倍，p=0.0069）在 Aagenaes 综合征患者出生时与对照者相比表达上调。
miRNA 不受多点采样或采血位置的影响，在多次冻融循环中稳定。新生儿筛查卡中的干血斑 miRNA 可用于检测 Aagenaes 综合征新生儿与健康对照者的 miRNA 差异表达。干血斑可能是探索循环 miRNA 作为生物标志物的有用来源。
循环 miRNA 可用作有用的生物标志物。DBS 中的 miRNA 不受多点采样或采血位置的影响，在多次冻融循环中稳定。即使个体数量较少，也允许区分患者和对照者。在出生后早期，胆汁淤积性淋巴水肿综合征患者表现出与肝纤维化相关的 miRNA 谱。本研究表明，新生儿筛查卡可能是研究 miRNA 的有用来源，因为技术变异性小于生物学变异性。
