PHYLOGENOMIC, TRANSCRIPTOMIC, VIROMIC, AND IMMUNOPROTEOMIC DETERMINANTS OF NECROTIZING ENTEROCOLITIS

项目摘要 极低出生体重儿的坏死性小肠结肠炎仍然是一个主要的挑战。我们的反应性临床 40年来，坏死性小肠结肠炎的治疗方法几乎没有变化。因以下原因引起的发病率和死亡率 坏死性小肠结肠炎的发病率仍然高得令人无法接受，预防策略依赖于有限的、 如果有的话，就是疗效。我们收集了大量的标本，并附有丰富的临床元数据， 以此来确定早产儿坏死性小肠结肠炎发生的机制 婴儿。我们的建议扩展了我们最近发表的发现，即肠道细菌失调，其特征是 伽马蛋白细菌的过度表达和专性厌氧细菌的表达不足， 在坏死性小肠结肠炎的发展之前，并看似有助于其发展。 我们的主要假设是，可识别的微生物和/或宿主机制意味着婴儿在 有坏死性小肠结肠炎的风险。我们的长期目标是利用我们对微生物驱动的知识 坏死性小肠结肠炎的病理生理学研究 细菌种群，以在突然出现坏死性病变之前表征宿主生物学 小肠结肠炎。通过生成这些数据，我们希望减轻坏死性小肠结肠炎的严重程度，或预防 总而言之。 我们的具体目标是：(1)在对肠道细菌进行全面的基因组分析之前 坏死性小肠结肠炎的发病，以及(2)定义坏死化前的肠道生物学和宿主反应 小肠结肠炎发作。所有病例的样本都将是在此事件发生之前获得的样本。控制 材料将与病例相匹配，根据出生时的胎龄和生命天数分别采集标本 制作。 为了实现这些目标，我们将使用我们的会前粪便和血清/血浆档案。 来自三个新生儿重症监护室的低出生体重儿。其中46名婴儿经历了NEC， 在排除了那些也患有先天性心脏病的人之后。我们将利用基因组特征来描述 协同分析中的分离、转录组、代谢组学、病毒组和免疫蛋白质组学 坏死性小肠结肠炎背后的微生物和宿主生物学。我们还将审问细菌的种群 与预防坏死性小肠结肠炎有关。所有研究都将首先分析来自 圣路易斯，然后在俄克拉何马城和路易斯维尔的标本中验证发现。这些病例对照和 时间序列比较将提供对棘手和高度影响的 早产并发症，使用一个特别相关的系统，即被殖民的人类婴儿。 到项目结束时，我们将生成机械信息数据，以指导尝试更好地 预测、治疗或理想情况下预防坏死性小肠结肠炎，这是早产的一种毁灭性并发症。

Project Summary Necrotizing enterocolitis in very low birth weight infants remains a major challenge. Our reactive clinical approach to necrotizing enterocolitis has changed little in four decades. Morbidity and mortality from necrotizing enterocolitis remain unacceptably high, and preventive strategies rely on interventions with limited, if any, efficacy. We have assembled an extensive set of specimens, accompanied by rich clinical metadata, with which to define mechanisms that underlie the development of necrotizing enterocolitis in very preterm infants. Our proposal extends our recently published findings that a gut bacterial dysbiosis, characterized by over-representation of Gammaproteobacteria and under-representation of obligate anaerobic bacteria, precede, and plausibly contribute to, the development of necrotizing enterocolitis. Our overarching hypothesis is that identifiable microbial and/or host mechanisms signify an infant at risk for necrotizing enterocolitis. Our long-term goals are to use our knowledge of microbial-driven pathophysiology in necrotizing enterocolitis to build interventions to prevent the assembly of dysbiotic gut bacterial populations, to characterize the host biology prior to the sudden appearance of necrotizing enterocolitis. By generating these data, we hope to lessen the severity of necrotizing enterocolitis, or to prevent it altogether. Our Specific Aims are: (1) Conduct a comprehensive genomic analysis of gut bacteria prior to necrotizing enterocolitis onset, and (2) Define the gut biology and host response prior to necrotizing enterocolitis onset. All specimens from cases will be those obtained before this event occurred. Control materials will be matched to cases, according to gestational age at birth, and day of life specimens were produced. To accomplish these Aims, we will use our archive of pre-event stools and sera/plasmas from 977 very low birthweight infants from three neonatal intensive care units. Forty-six of these infants experienced NEC, after excluding those who also had congenital heart disease. We will employ genomic characterization of isolates, transcriptomics, metabolomics, viromics, and immunoproteomics in coordinated analyses to illuminate microbial and host biology underlying necrotizing enterocolitis. We will also interrogate populations of bacteria associated with protection from necrotizing enterocolitis. All studies will initially analyze a primary cohort from St. Louis, and then validate findings in specimens from Oklahoma City and Louisville. These case-control and time-series comparisons will provide mechanistic insight into an intractable and highly consequential complication of preterm birth, using an exceptionally relevant system, i.e., the colonized human infant. By project conclusion, we will generate mechanistically-informed data to guide attempts to better anticipate, treat, or, ideally, prevent, necrotizing enterocolitis, a devastating complication of preterm birth.