Microfluidic Assessment of Clinical Outcomes in Preterm Newborns

败血症对早产(早产)人群的影响最大。新生儿败血症(新生儿内败血症 生命第一个月)每年导致全球100多万人死亡，是最常见、最困难的疾病之一 和昂贵的问题来诊断、治疗和预防。早产儿患败血症的几率高达1000倍。 高于足月儿，首当其冲的是相关死亡率和终生败血症--幸存者 发病率。潜在感染新生儿的处理中存在的实质性临床问题 未回答：1)婴儿感染了吗？2)哪些婴儿有复杂临床病程的最大风险 感染？以及3)为什么早产儿(特别是极低出生体重儿)有如此高的风险 发展中的感染？对早产儿研究的缺乏在很大程度上可以归因于 用于研究的有限的血量(典型的28周的总血量为80-100毫升，1000克婴儿)，以及 先前的假设是，新生儿宿主的免疫反应类似于年龄较大的儿童和成年人。 我们认为新生儿的中性粒细胞(PMN)先天免疫功能缺陷是其潜在的原因之一。 这种宿主保护性免疫力下降的原因，其功能可以用来预测 脓毒症和旷日持久的临床病程。我们打算提供准确的方法来诊断脓毒症，识别 预后和危重疾病分层标志物，并发现免疫差异与潜在的 可能改善新生儿感染相关结局的转化性干预，所有这些都基于一种新的 微流控平台。具体地说，我们提出了一项针对300名早产儿(30周)的前瞻性、观察性研究 胎龄)和60名足月(&gt；36周)婴儿，我们将在这些婴儿中发现PMN的具体缺陷 功能，开发脓毒症的预测模型，最终，基于微流体的临床结果 中性粒细胞功能和转录组的测量，以及经典的临床测量。该项目由 几种新的、经过验证的微流控技术，它们坚固耐用，只需很少的培训就能轻松使用。这些 技术为血液PMN群体的功能提供了全面的衡量标准；关键的细胞 先天免疫的组成部分。我们还将从微流控分选的PMN中提取高质量的核酸 转录分析，并提供完整的血细胞计数和5部分差异与临床相结合 化学和炎症生物标记物。总的来说，这些技术总共需要~100微升(L) 血液，这使得它们对样本量有限的早产儿特别有用，并有助于 对PMN的关键职能进行了前所未有的时间分辨率的系列评估。这些研究，综合 利用生物信息学方法，将产生新的工具来诊断新生儿败血症并预测 临床结果。这种方法有能力极大地改变 早产儿，并有可能改善长期结果，同时降低医院成本。

Sepsis has its greatest impact in the prematurely born (preterm) population. Neonatal sepsis (sepsis within the first month of life) causes over one million deaths worldwide annually, and is one of the most common, difficult and costly problems to diagnose, treat and prevent. The preterm infant can suffer rates of sepsis up to 1000-fold higher than the full-term infant, and bears the brunt of the associated mortality and lifelong sepsis-survivor morbidity. Substantial clinical questions in the management of the potentially infected neonate remain unanswered: 1) Is the infant infected?, 2) Which infants have the greatest risk for a complicated clinical course with infection?, and 3) Why are preterm infants (especially very low birth weight infants) at such high risk of developing infections? The paucity of investigations in the preterm infant can be attributed in large part to very limited blood volume for study (80-100 milliliter total blood volume in a typical 28 week, 1000 gram infant), and prior assumptions that the neonatal host immune response is similar to that seen in older children and adults. We propose that deficiencies in innate immune function of neutrophils (PMN) in neonates are one underlying cause of this decrease in host protective immunity, and their function can be used to predict the development of sepsis and protracted clinical course. We intend to deliver accurate methods to diagnose sepsis, identify prognostic and critical illness stratification markers, and uncover immunological differences with the potential for translational interventions that may improve neonatal infection-related outcomes, all based on a novel microfluidics platform. Specifically, we propose a prospective, observational study of 300 preterm (<30 weeks gestational age) and 60 full-term (>36 weeks) infants in whom we will identify specific deficiencies in PMN function, develop prediction models for sepsis, and ultimately, clinical outcome, based on microfluidics measurements of PMN function and transcriptomics, and classical clinical measures. The project is enabled by several novel, validated, microfluidic technologies that are robust and easy to use with little training. These technologies provide comprehensive measures of the functionality of blood PMN population; a critical cellular component of innate immunity. We will also extract high-quality nucleic acids from microfluidic-sorted PMNs for transcriptomic analyses, and deliver a complete blood count with 5-part differential integrated with clinical chemistry and inflammatory biomarkers. Collectively, these techniques require a total of ~100 microliters (L) of blood, which makes them particularly useful for preterm infants where sample volume is limited, and facilitates serial assessments with unprecedented temporal resolution of key functions of PMNs. These studies, integrated with bioinformatics approaches, will generate new tools for diagnosing sepsis in the newborn and predicting clinical outcomes. Such approaches have the capability to dramatically change the clinical management of the preterm infant, and potentially improve long-term outcomes while reducing hospital costs.