Genomic and Epigenomic Mechanisms of Maternal E-Cigarette induced Abnormal Brain Development

项目摘要/摘要 母亲吸烟是一个公认的公共卫生问题，与神经发育增加有关 子代的疾病和其他健康风险。大量证据表明，母婴的各种不良影响 吸烟或产前尼古丁暴露对新生儿大脑发育的影响，例如，多动，认知能力下降 表现，增加焦虑和抑郁，增加对脑损伤的易感性。电子烟 (e-cig)在孕妇和青少年中很受欢迎。越来越多的证据表明 母亲使用电子烟会影响大脑发育，导致皮质神经元形态异常和异常。 子代的神经行为。使用大鼠出生前暴露于e-cig的模型和单核测序，我们 最近发现，产前接触电子烟破坏了兴奋性-抑制性(E/I)平衡，兴奋性/抑制性(E/I)比值 新生儿大脑中的抑制性神经元。E/I平衡对于正常的大脑发育和功能至关重要，而且 它的破坏被认为是许多神经发育障碍的基础，包括 自闭症谱系障碍、多动症和其他精神障碍。然而，潜在的基因组和 表观基因组学机制尚不清楚。这项调查旨在填补这一知识空白。我们的假设 产前暴露e-cig可诱导发育过程中的表观基因组重编程和基因组改变 这会导致E/I失衡，从而增加神经发育障碍的风险。 利用我们最近开发的产前e-CIG暴露模型，我们提出了三个具体目标。目标1将 确定产前电子烟暴露对大脑发育和E/I进展的时空影响 使用空间基因组学和单细胞测序技术的不平衡。目标2将确定表观基因组 胎儿期E-CIG的调节机制导致E/I失衡。目标3将调查产前电子烟是否 导致涉及E/I失衡的基因组改变(即SNV和CNV)。许多尖端技术 在我们的研究中利用了基因组学和表观基因组学技术，包括空间基因组学、单核 Rna-seq和染色质可获得性(snatac-seq)，以及最先进的生物信息学工具。结果：我们的 研究将：1)确定由产前电子烟暴露引起的E/I失衡的大脑区域；2)确定 产前e-CIG诱发E/I的关键基因、基因组变化及表观基因组调控 3)产生了大量关于产前暴露于电子烟的大鼠大脑的前所未有的组学数据。 影响：使用成熟的宫内e-CIG暴露模型，结合尖端基因组学和 表观基因组学方法，我们建议的研究将阐明基因组和表观基因组学机制 潜在的母体电子烟导致大脑发育异常，为研究其影响提供了有价值的新见解 E-cig对早期中枢神经系统发育的影响，这将有助于探索有前途的分子和 治疗电子烟蒸发所致脑损伤的细胞治疗靶点。

PROJECT SUMMARY/ABSTRACT Maternal smoking is a well-recognized public health concern associated with increased neurodevelopmental disorders and other health risks in offspring. Substantial evidence indicates various adverse effects of maternal smoking or prenatal nicotine exposure on neonatal brain development, e.g., hyperactivity, reduced cognitive performance, increased anxiety and depression, and increased susceptibility to brain injury. Electronic cigarettes (e-cig) have become popular in pregnant women and young adolescents. Growing evidence suggests that maternal e-cig use affects brain development, resulting in abnormal cortical neuronal morphology and aberrant neuro-behaviors in offspring. Using a rat prenatal e-cig exposure model and single-nucleus sequencing, we recently found that prenatal e-cig exposure disrupted excitatory-inhibitory (E/I) balance, ratio of excitatory/ inhibitory neurons, in the neonatal brain. E/I balance is crucial for normal brain development and functions, and its disruption has been postulated to underlie the pathogenesis of many neurodevelopmental disorders, including autism spectrum disorders, ADHD, and other psychiatric disorders. However, the underlying genomic and epigenomic mechanisms are still not known. This investigation seeks to fill this knowledge gap. Our hypothesis is that prenatal e-cig exposure induces epigenomic reprogramming and genomic alterations in the developing brain, which cause an E/I imbalance and consequently an increased risk for neurodevelopmental disorders. Exploiting the prenatal e-cig exposure model we developed recently, we propose three specific aims. Aim 1 will determine the spatial-temporal effects of prenatal e-cig exposure on brain development and progression of E/I imbalance using spatial genomics and single-cell sequencing techniques. Aim 2 will determine the epigenomic mechanisms regulating the prenatal e-cig induced E/I imbalance. Aim 3 will investigate whether prenatal e-cig causes genomic alterations (i.e., SNVs and CNVs) that are involved in E/I imbalance. Many cutting-edge genomic and epigenomic technologies are exploited in our studies, including spatial genomics, single-nucleus RNA-seq and chromatin accessibility (snATAC-seq), and state-of-the-art bioinformatics tools. OUTCOMES: Our study will: 1) identify the brain regions that show the E/I imbalance caused by prenatal e-cig exposure; 2) identify the key genes, genomic changes and epigenomic regulations responsible for the prenatal e-cig induced E/I imbalance; 3) produce a large amount of unprecedented omics data on the rat brain prenatally exposed to e-cig. IMPACT: Using a well-established intrauterine e-cig exposure model coupled with cutting-edge genomics and epigenomics approaches, our proposed studies will elucidate the genomic and epigenomic mechanisms underlying maternal e-cig induced abnormal brain development, providing valuable new insights into the effects of e-cig on the early central nervous system development, which will help explore promising molecular and cellular therapeutic targets for treating e-cig vaping-induced brain damage.