Prenatal Traffic-Related Air Pollutants, Placental Epitranscriptomics, and Child Cognition

项目摘要 空气污染（TRAP）是一种普遍存在的环境暴露， 在动物和人类研究中对神经发育的不良影响。然而，这些的确定机制 影响目前尚不清楚，限制了我们的生物学理解，并延迟了介入和治疗 努力保护儿童免受这种广泛的接触。胎盘负责产前神经发育 通过调节胎儿生长及其神经内分泌功能，一直被认为是一种 TRAP对发育中的大脑的影响的主要介质。RNA的转录后修饰， 也就是说，表转录组对环境敏感且对胎盘发育至关重要， 功能，因此提供了一个尚未探索的途径，以确定新的机制的TRAP神经毒性。N6- 甲基腺苷（m6 A）是信使RNA（mRNA）上最普遍的表位转录组修饰， mRNA剪接、稳定性和翻译的调节因子。m6 A阅读器，写入器和擦除器（RWE）蛋白， 从mRNA中解释、添加和去除m6 A标记也对毒物高度敏感。我们团队最近 表明TRAP到达人胎盘，从而表明局部TRAP积累可能直接影响胎盘的功能。 改变胎盘的表型和功能。然而，迄今为止还没有研究调查过 胎盘暴露于TRAP和m6 A表观转录组学之间的关系，也没有任何以前的研究 研究TRAP诱导的m6 A表转录组学改变与未来儿童神经发育的关系。 我们假设胎盘TRAP负荷通过胎盘中TRAP的变化对认知产生不利影响。 m6 A表位转录组。为了研究这一假设，我们提出了一系列协调的人类和体外 问题研究对于人体研究，我们将利用位于纽约的哥伦比亚中心的两个队列， -Hermanos队列，作为发现和复制 集.在目的1中，我们将鉴定人胎盘中与产前诊断相关的m6 A表位转录组改变， 陷阱我们将使用一种创新的方法来量化直接在胎盘上的BC和环境空气污染 整个孕期的评估。我们将使用m6 A测序来分析胎盘m6 A并测量蛋白质 18例胎盘m6 A RWEs的mRNA表达。我们将使用数据驱动的方法来识别生物 TRAP相关胎盘功能障碍的相关通路。在目标2中，我们将鉴定m6 A表转录组， 人类胎盘的改变与5-6岁儿童的认知能力有关， 标准的神经心理测试在目标3中，我们将进行滋养层细胞的体外研究以鉴定TRAP-1。 诱导m6 A表位转录组的改变及其对mRNA稳定性、剪接和翻译的影响。 总之，这些目标将揭示TRAP对胎盘epitranscriptome的影响，以及随之而来的 对儿童神经发育的影响这项研究将导致新的见解的机制，连接 产前环境对儿童神经发育的影响

PROJECT SUMMARY Traffic-related air pollution (TRAP) is a ubiquitous environmental exposure that has been consistently linked to adverse neurodevelopmental effects in animal and human studies. However, definitive mechanism(s) for these effects is currently unknown, limiting our biological understanding and delaying interventional and therapeutic efforts to protect children from this widespread exposure. The placenta oversees prenatal neurodevelopment through regulation of fetal growth and its neuroendocrine functions and has been consistently indicated as a primary mediator of the effects of TRAP on the developing brain. Post-transcriptional modifications of RNA, i.e., the epitranscriptome, are both environmentally sensitive and critical to placental development and functions, hence providing a yet unexplored avenue to identify new mechanisms of TRAP neurotoxicity. N6- methyladenosine (m6A) is the most prevalent epitranscriptomic modification on messenger RNA (mRNA) and a regulator of mRNA splicing, stability, and translation. The m6A reader, writer, and eraser (RWE) proteins that interpret, add, and remove m6A marks from mRNA are also highly sensitive to toxicants. Our team recently showed that TRAP reaches the human placenta, thereby indicating that local TRAP accumulation may directly alter the placental epitranscriptome and function. However, no research to date has investigated the relationships between placental exposure to TRAP and m6A epitranscriptomics, nor has any previous study investigated TRAP-induced m6A epitranscriptomic alterations in relation to future child neurodevelopment. We hypothesize that placental TRAP load adversely impacts cognition via changes in the placental m6A epitranscriptome. To investigate this hypothesis, we propose a coordinated series of human and in vitro studies. For the human studies, we will leverage two cohorts within the NYC-based Columbia Center for -Hermanos cohorts, as discovery and replication sets. In Aim 1, we will identify m6A epitranscriptome alterations in human placenta associated with prenatal TRAP. We will use an innovative method to quantify BC directly on the placenta and ambient air pollution assessment throughout pregnancy. We will use m6A-sequencing to profile placental m6A and measure protein and mRNA expression of 18 placental m6A RWEs. We will use data-driven approaches to identify biological pathways implicated in TRAP-related placental dysfunction. In Aim 2, we will identify m6A epitranscriptome alterations in human placenta associated with child cognition at ages 5-6 measured through a battery of gold- standard neuropsychological tests. In Aim 3, we will conduct in vitro studies of trophoblasts to identify TRAP- induced alterations of the m6A epitranscriptome and their impact on mRNA stability, splicing, and translation. Together, these aims will uncover the impacts of TRAP on the placental epitranscriptome and the ensuing effects on child neurodevelopment. This research will lead to novel insight into the mechanisms linking the prenatal environment to child neurodevelopment.