Siddaiah Roopa, Emery Lucy, Stephens Heather, Donnelly Ann, Erkinger Jennifer, Wisecup Kimberly, Hicks Steven D, Kawasawa Yuka Imamura, Oji-Mmuo Christiana, Amatya Shaili, Silveyra Patricia
Department of Pediatrics, Penn State Health Children's Hospital, Hershey, PA 17036, USA.
Penn State Health College of Medicine, Hershey, PA 17036, USA.
Life (Basel). 2022 Mar 30;12(4):506. doi: 10.3390/life12040506.
Background: MicroRNAs (miRNA) are small non-coding RNAs that regulate gene expression playing a key role in organogenesis. MiRNAs are studied in tracheal aspirates (TA) of preterm infants. However; this is difficult to obtain in infants who are not intubated. This study examines early salivary miRNA expression as non-invasive early biomarkers in extremely low gestational age newborns (ELGANs). Methods: Saliva was collected using DNA-genotek swabs, miRNAs were analyzed using RNA seq and RT PCR arrays. Salivary miRNA expression was compared to TA using RNA seq at 3 days of age, and longitudinal changes at 28 days of age were analyzed using RT PCR arrays in ELGANs. Results: Approximately 822 ng of RNA was extracted from saliva of 7 ELGANs; Of the 757 miRNAs isolated, 161 miRNAs had significant correlation in saliva and TA at 3 days of age (r = 0.97). Longitudinal miRNA analysis showed 29 miRNAs downregulated and 394 miRNAs upregulated at 28 days compared to 3 days of age (adjusted p < 0.1). Bioinformatic analysis (Ingenuity Pathway Analysis) of differentially expressed miRNAs identified organismal injury and abnormalities and cellular development as the top physiological system development and cellular function. Conclusion: Salivary miRNA expression are source for early biomarkers of underlying pathophysiology in ELGANs.
微小RNA（miRNA）是一类小的非编码RNA，可调节基因表达，在器官发生中起关键作用。已在早产儿的气管吸出物（TA）中对miRNA进行了研究。然而，对于未插管的婴儿而言，获取气管吸出物很困难。本研究检测极早早产儿（ELGANs）唾液中miRNA的早期表达，将其作为非侵入性早期生物标志物。方法：使用DNA Genotek拭子收集唾液，通过RNA测序和逆转录聚合酶链反应（RT PCR）阵列分析miRNA。在出生3天时，使用RNA测序比较ELGANs唾液中miRNA的表达与气管吸出物中的表达，并在出生28天时，使用RT PCR阵列分析ELGANs中miRNA的纵向变化。结果：从7例ELGANs的唾液中提取了约822 ng的RNA；在分离出的757种miRNA中，有161种miRNA在出生3天时唾液和气管吸出物中的表达具有显著相关性（r = 0.97）。纵向miRNA分析显示，与出生3天时相比，出生28天时29种miRNA表达下调，394种miRNA表达上调（校正p < 0.1）。对差异表达的miRNA进行生物信息学分析（Ingenuity通路分析），确定机体损伤和异常以及细胞发育是首要的生理系统发育和细胞功能。结论：唾液中miRNA的表达是ELGANs潜在病理生理学早期生物标志物的来源。
