Maternal circulating miRNA function in Fetal Alcohol Spectrum Disorders

项目摘要 产前酒精暴露（PAE）是智力和其他大脑残疾的主要非遗传原因。 然而，胎儿酒精谱系障碍（FASD）估计影响美国约2 - 5%的学龄儿童， 仍然难以诊断和预防。我们最近的工作（PMCID 5102408）确定了几个循环 重度饮酒孕妇中的microRNAs（miRNAs），其在第2和第3天的表达水平 孕三个月可预测婴儿的不良结局，包括颅面畸形、神经行为和生长 赤字1.具体而言，方差分析（ANOVA）模型鉴定了母亲中11种升高的血浆miRNA 他们的婴儿受到酒精消费的严重影响。使用随机森林分析（RFA），我们 进一步能够使用一组不同的miRNA对明显未受乙醇暴露影响的婴儿进行分类， 受影响的婴儿与未接触酒精的婴儿相比。除了它们的诊断价值，还不知道这些 与不良婴儿结局相关的母体循环miRNA（M-circmiRNA-aAIO）有助于 FASD的发展病理学。生物信息学分析表明，这些循环的miRNA可能 调节STAT3和ephrin信号通路的重要枢纽基因，这些基因反过来又控制周期 上皮间质转化（EMT）对正常胚胎发生、胎盘和胎儿至关重要 发展因此，我的总体假设是，PAE通过干扰 循环母体miRNAs对胎盘和胎儿的内分泌作用。在我的研究计划中， 使用人类细胞系和体内小鼠模型，我将测试的假设，致病水平的 循环miRNA介导乙醇对胎盘和胎儿的影响，它们控制胚胎生长， 通过调节关键胎盘和胎儿细胞的EMT样行为来维持存活。该项目的长期目标将 开发干预策略，利用内分泌循环miRNAs的生物学， 由于PAE和其他致畸剂导致的不良结果。我的近期目标将是定义母亲的角色， 预测FASD结果的miRNA对胎盘和胚胎生长和发育的影响。 目的1：确定M-circmiRNA-aAIO对滋养层细胞生长、存活、迁移和侵袭的影响。 基础和乙醇暴露条件。 目的2：确定M-circmiRNA-aAIO对胚胎生长、死亡和细胞凋亡的影响。 在基础和乙醇暴露条件下的成熟/分化。 目的3：确定M-circmiRNA-aAIO对体内胎儿发育和新生儿结局的影响。 我的研究有望揭示FASD相关的新的内分泌和妊娠相关功能， 母体系统性miRNAs，并可能提供新的治疗靶点和非侵入性方式，以减轻 PAE的影响。考虑到它的转化潜力，这个项目将进一步培养我作为一个医生科学家 对从事临床工作和早期发育障碍研究感兴趣。

Project Summary Prenatal alcohol exposure (PAE) is the leading non-genetic cause of intellectual and other brain disabilities. However, fetal alcohol spectrum disorders (FASD), estimated to affect ~2-5% of school-aged children in the US, remain difficult to diagnose and to prevent. Our recent work (PMCID 5102408) identified several circulating microRNAs (miRNAs) in heavy alcohol-consuming pregnant women whose expression levels in the 2nd and 3rd trimester predicted adverse infant outcomes including craniofacial anomalies and neurobehavioral and growth deficits1. Specifically, Analysis of Variance (ANOVA) models identified 11 elevated plasma miRNAs in mothers whose infants were severely affected by alcohol consumption. Using Random Forest Analysis (RFA), we were further able to use a distinct group of miRNAs to classify infants apparently unaffected by ethanol exposure with affected infants as opposed to alcohol unexposed infants. Aside from their diagnostic value, it is unknown if these Maternal circulating miRNAs associated with Adverse Infant Outcomes (M-circmiRNA-aAIO) contribute to the developmental pathologies of FASD. Bioinformatic analysis suggests these circulating miRNAs potentially regulate important hub genes for STAT3 and ephrin signaling pathways, which are in turn known to control cycles of epithelial mesenchymal transition (EMT) crucial for normal embryogenesis and placental and fetal development. Therefore, my overarching hypothesis is that PAE impairs early development by interfering with the endocrine action of circulating maternal miRNAs on both the placenta and fetus. In my research proposal, using both human cell lines and in vivo mouse models, I will test the hypothesis that pathogenic levels of circulating miRNAs mediate effects of ethanol on placenta and fetus and that they control embryo growth and survival by regulating EMT-like behaviors of key placental and fetal cells. The long-term goals of this project will be to develop interventional strategies that exploit the biology of endocrine circulating miRNAs in mitigating negative outcomes due to PAE and other teratogens. My immediate goals will be to define the role of maternal miRNAs, which predict FASD outcomes, on placental and embryo growth and development. Aim 1: Determine the impact of M-circmiRNA-aAIOs on trophoblast growth, survival, migration and invasion under basal and ethanol exposed conditions. Aim 2: Determine the impact of M-circmiRNA-aAIOs on embryonic growth, death, and cellular maturation/differentiation under basal and ethanol exposed conditions. Aim 3: Determine the impact of M-circmiRNA-aAIOs on in vivo fetal development and neonatal outcomes. My studies are expected to uncover novel endocrine and pregnancy related functions of FASD-associated maternal systemic miRNAs and may provide novel therapeutic targets and non-invasive modalities to mitigate effects of PAE. Given its translational potential, this project will further my training as a physician-scientist interested in pursuing both clinical duties and research on early developmental disorders.