PCR-based miRNA assessment of neonatal opioid withdrawal syndrome (NOWS) severity

摘要 阿片类药物使用障碍（OUD）已经成为一个全国性的危机，影响到200多万美国人。的数量 在过去的十年中，美国患有OUD的孕妇增加了近5倍，导致了类似的激增。 新生儿阿片类药物戒断综合征（NOWS）婴儿，从1.5至8每1,000名医院分娩（32000 2004年至2014年期间的案件总数。相关的医院费用增加了35%，达到5.32亿美元， 2014年（其中4.62亿美元由医疗补助计划资助）。在产前暴露于非法阿片类药物的新生儿中， 55 - 94%的人出现戒断症状，包括呼吸、胃肠道和喂养问题。婴幼儿 严重的NOWS通常需要延长住院时间，2 - 11%的患者出现严重并发症，如 癫痫发作和体重减轻严重的病例通常需要药物干预以减轻长期的负性- 长期影响婴儿的神经发育。根据国家指南，阿片类药物使用者所生的婴儿 在医院监测（72 - 96小时）NOWS体征。然而，目前使用不同的比例尺， 监测症状的严重程度，导致诊断不一致。此外，NOWS可以呈现 从出生到7天后，有时会导致释放后昂贵的重新入院。当 需要采取药物措施，这些延误可能会导致婴儿发病的风险增加。因此，我们认为， 迫切需要一种可获得的、廉价的、灵敏的和准确的诊断工具，以支持最佳的 NOWS婴儿的临床结局。为了满足这一需求，FYR Diagnostics与University of 新墨西哥州（UNM）和得克萨斯A & M大学（TAMU），将开发一种方便，非侵入性 脐带样本的诊断分析，用于早期检测严重的NOWS，以改善 确定高危和低危婴儿。该分析将基于两项创新：a）一种miRNA， 识别患有严重NOWS的婴儿的特征;和B）专利申请中的超灵敏DNA 扩增反应（UDAR）化学，使用简单的引物设计快速直接检测单个 目前低浓度的miRNAs低于检测限。在第一阶段，FYR将展示 这种新型定量工具的可行性：1）在预测性miRNA上测试专有UDAR化学 使用UNM/TAMU先前使用的脐带血浆样本的生物标志物;和2）比较UDAR- 用于检测NOWS miRNA生物标志物的基于RT-qPCR的传统测定法，以验证测定法 性能，并证明基于UDAR的检测试剂盒具有等同或上级特异性， 灵敏度在第二阶段，FYR，UNM和TAMU将进行大规模临床研究以验证NOWS 早期检测工具临床验证研究（N = 150）将评价 基于UDAR的测定，证明其表现出与NOWS相当的性能和预测效用 OUD女性所生婴儿的临床变异性的严重程度。该项目将产生关键的 FDA批准多中心试验所需的数据和商业化工作的基础。

Abstract Opioid use disorder (OUD) has become a national crisis, affecting over 2 million Americans. The number of pregnant women with OUD in the U.S. increased almost 5-fold in the past decade, leading to a similar surge in infants born with neonatal opioid withdrawal syndrome (NOWS), from 1.5 to 8 per 1,000 hospital births (32,000 cases total) between 2004 and 2014. Associated hospital costs have risen by 35%, reaching $532 million in 2014 ($462 million of which was financed by Medicaid). Among neonates prenatally exposed to illicit opioids, 55-94% develop withdrawal symptoms, including respiratory, gastrointestinal, and feeding problems. Infants with severe NOWS often require prolonged hospitalization, and 2-11% develop serious complications, such as seizures and weight loss. Severe cases often require pharmacological intervention to mitigate negative long- term impacts on the infant's neurological development. Per national guidelines, infants born to opioid users are monitored (72-96 hours) in the hospital for signs of NOWS. However, different scales are currently in use to monitor the severity of symptoms, leading to inconsistencies in diagnosis. Moreover, NOWS can present anywhere from birth to 7 days after, sometimes leading to costly readmissions after release. When pharmacologic measures are needed, these delays can lead to an increased risk of infant morbidity. Therefore, there is an urgent need for an accessible, inexpensive, sensitive, and accurate diagnostic tool to support optimal clinical outcomes of infants with NOWS. To meet this need, FYR Diagnostics, alongside the University of New Mexico (UNM) and Texas A&M University (TAMU), will develop a convenient, non-invasive diagnostic assay from umbilical cord samples for early detection of severe NOWS to improve identification of high- and low-risk infants. The assay will be based on two innovations: a) an miRNA signature that identifies infants with severe NOWS; and b) a proprietary patent-pending ultrasensitive DNA Amplification Reaction (UDAR) chemistry that uses a simple primer design to rapidly and directly detect individual miRNAs at low concentrations currently below the limit of detection. In Phase I, FYR will demonstrate the feasibility of this novel quantitative tool by 1) testing the proprietary UDAR chemistry on predictive miRNA biomarkers using umbilical cord plasma samples previously used by UNM/TAMU; and 2) comparing the UDAR- based assay to traditional RT-qPCR-based assays for detection of NOWS miRNA biomarkers to validate assay performance and demonstrate that the UDAR-based assay exhibits equivalent or superior specificity and sensitivity. In Phase II, FYR, UNM, and TAMU will conduct a large-scale clinical study to validate the NOWS early detection tool. The clinical validation study (N=150) will evaluate the analytical and clinical feasibility of the UDAR-based assay, demonstrating that it exhibits comparable performance and predictive utility of NOWS severity across the clinical variability of infants born to women with OUD. The project will generate the pivotal data necessary for FDA approval of a multi-center trial and a basis for commercialization efforts.