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

摘要 阿片类药物使用障碍(OUD)已成为一种全国性的危机，影响着200多万美国人。数量 在过去的十年里，美国患有多发性硬化症的孕妇增加了近5倍，导致了类似的激增 出生时患有新生儿阿片类药物戒断综合征(NOWS)的婴儿，从每1,000名住院分娩中1.5人增加到8人(32,000人) 病例总数)。相关医院成本上涨了35%，年内达到5.32亿美元 2014年(其中4.62亿美元由医疗补助提供资金)。在产前接触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。婴儿患有 严重的NOW通常需要长期住院，2%-11%的人会出现严重的并发症，如 癫痫发作和体重减轻。危重病例往往需要药物干预来缓解长期的负面影响。 对婴儿神经发育的影响。根据国家指南，阿片类药物使用者所生婴儿 在医院监测(72-96小时)NOWS的迹象。然而，目前使用不同的比例尺来 监控症状的严重程度，导致诊断不一致。此外，NOWS可以提供 从出生到7天后的任何地方，有时会导致出院后昂贵的重新入院。什么时候 需要采取药物措施，这些延误可能会导致婴儿发病风险增加。因此， 迫切需要一种可访问、廉价、灵敏和准确的诊断工具来支持最佳 新生儿NOWS的临床转归。为了满足这一需求，FYR诊断公司与加州大学 新墨西哥大学(UNM)和德克萨斯农工大学(TAMU)将开发一种方便、非侵入性的 对脐带标本进行早期诊断以提高重度NOWS诊断水平 高危和低危婴儿的识别。这项检测将基于两项创新：a)miRNA 识别患有严重NOWS的婴儿的签名；以及b)正在申请专利的超敏感DNA 扩增反应(UDAR)化学，使用简单的引物设计快速、直接地检测个体 目前低于检测限值的低浓度miRNAs。在第一阶段，FYR将展示 这个新的定量工具的可行性通过1)测试预测miRNA的专利UDAR化学 使用UNM/TAMU以前使用的脐带血浆样本的生物标记物；以及2)比较UDAR- 以传统RT-qPCR为基础的检测NOWS miRNA生物标记物的方法验证分析 性能，并证明了基于UDAR的检测显示出同等或更好的特异性和 敏感度。在第二阶段，FYR、UNM和TAMU将进行一项大规模的临床研究，以验证NOWS 早期检测工具。临床验证研究(N=150)将评估分析和临床可行性 基于UDAR的检测，表明其表现出与NOWS相当的性能和预测效用 OUD妇女所生婴儿的临床变异性的严重程度。该项目将生成Pivotal FDA批准多中心试验所需的数据，并为商业化努力奠定基础。

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.