Supplementary Material for: MicroRNA Expression Profiles as Diagnostic and Prognostic Biomarkers of Perinatal Asphyxia and Hypoxic-Ischaemic Encephalopathy

<b><i>Introduction:</i></b> Perinatal asphyxia is a leading cause of neonatal death. Up to one-third of asphyxiated neonates suffer from hypoxic-ischaemic encephalopathy (HIE) with substantial long-term morbidity. Currently available diagnostic and prognostic tools bear limitations, and additional reliable biomarkers are needed for all stages of clinical management. A novel tool in neuroscientific research is micro-ribonucleic acid (miRNA) profiling. The aim of the present study was to determine miRNA expression profiles of healthy and asphyxiated neonates with and without HIE and to assess their potential as diagnostic and prognostic biomarkers. <b><i>Methods:</i></b> We prospectively enrolled 49 neonates with a gestational age of ≥36 weeks, 15 of which fulfilled the diagnostic criteria of perinatal asphyxia and 34 served as healthy controls. Dried blood spots were collected from umbilical cord blood (UCB) and from venous blood upon admission to neonatal intensive care unit (NICU) and at 48 h of life. Samples were analysed by means of FirePlex™ technology (Abcam, Cambridge, MA, USA). <b><i>Results:</i></b> In the UCB, miRNA expression levels of hsa-mir-124-3p, hsa-mir-1285-5p, and hsa-mir-331-5p were significantly lower in asphyxiated neonates compared to healthy controls. Asphyxiated neonates requiring therapeutic hypothermia had significantly increased expression of hsa-miR-30e-5p and significantly decreased expression of hsa-miR-142-3p, hsa-miR-338-3p, hsa-miR-34b-3p, hsa-miR-497-5p, and hsa-miR-98-5p at the time of admission to the NICU. At 48 h, infants suffering from moderate/severe HIE with a poor long-term neurodevelopmental outcome showed a significant increase in hsa-mir-145-5p. <b><i>Discussion/Conclusion:</i></b> MiRNA profiling shows promise as a biomarker for perinatal asphyxia, hypothermia-requiring HIE, and poor neurodevelopmental outcome. Confirmatory studies are called for.

<b><i>引言：</i></b> 围产期窒息是新生儿死亡的首要诱因。多达三分之一的窒息新生儿会罹患缺氧缺血性脑病（hypoxic-ischaemic encephalopathy, HIE），并面临显著的长期致残风险。当前可用的诊断与预后工具均存在局限性，临床管理全阶段均亟需可靠的新型生物标志物。神经科学研究中的一项新兴技术为微核糖核酸（micro-ribonucleic acid, miRNA）表达谱分析。本研究旨在明确健康新生儿、伴或不伴HIE的窒息新生儿的miRNA表达谱，并评估其作为诊断与预后生物标志物的潜力。<b><i>方法：</i></b> 本研究前瞻性纳入49名胎龄≥36周的新生儿，其中15名符合围产期窒息的诊断标准，剩余34名作为健康对照。分别采集脐血（umbilical cord blood, UCB）的干血斑，以及新生儿入住新生儿重症监护病房（neonatal intensive care unit, NICU）时的静脉血干血斑与出生后48小时的静脉血干血斑。采用FirePlex™技术（美国马萨诸塞州剑桥市Abcam公司）对样本进行检测分析。<b><i>结果：</i></b> 在脐血样本中，窒息新生儿的hsa-mir-124-3p、hsa-mir-1285-5p及hsa-mir-331-5p的表达水平较健康对照显著降低。需要接受治疗性低温治疗的窒息新生儿，在入住NICU时，其hsa-miR-30e-5p的表达水平显著升高，而hsa-miR-142-3p、hsa-miR-338-3p、hsa-miR-34b-3p、hsa-miR-497-5p及hsa-miR-98-5p的表达水平显著降低。出生后48小时，罹患中重度HIE且长期神经发育结局不良的婴儿，其hsa-mir-145-5p的表达水平显著升高。<b><i>讨论与结论：</i></b> miRNA表达谱分析作为围产期窒息、需低温治疗的HIE以及不良神经发育结局的生物标志物具有应用前景，未来需开展更多验证性研究。