The Role of m6A-RNA Methylation in Memory Formation and Recall and Its Modulation and Influence on Long-Term Outcomes as a Consequence of Early Life Lead Exposure.

m6A-RNA 甲基化在记忆形成和回忆中的作用及其对早期铅暴露后果的长期结果的调节和影响。

• 批准号: 9927737
• 负责人: JAY S SCHNEIDER
• 金额: $ 16.41万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-15 至 2021-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9927737
• 关键词: Address; Adenosine; Adverse effects; Affect; Animal Model; Animals; Auditory; Basic Science; Behavioral; Brain; Brain region; Bypass; Chemicals; Child; Cognitive; Cues; DNA; DNA Methylation; Data; Development; Dimensions; Disease; Epigenetic Process; Fatty acid glycerol esters; Female; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Health; Hippocampus (Brain); Impaired cognition; Impairment; Lead; Life; Maintenance; Maps; Memory; Memory impairment; Messenger RNA; Methylation; Methyltransferase; Modification; Molecular; Mus; Neuronal Plasticity; Neurotoxins; Obesity; Outcome; Pathologic; Pathway interactions; Performance; Play; Post-Transcriptional Regulation; Prefrontal Cortex; Promoter Regions; Public Health; Publishing; RNA; RNA immunoprecipitation sequencing; RNA methylation; Rattus; Regulatory Pathway; Reporting; Research; Role; Site; Testing; Training; Transcript; Transcriptional Regulation; Translations; Work; adverse outcome; base; behavioral plasticity; behavioral response; brain behavior; conditioned fear; epitranscriptome; epitranscriptomics; experience; gene environment interaction; genetic regulatory protein; genome wide methylation; histone modification; improved; insight; knock-down; lead exposure; lentiviral-mediated; male; memory encoding; memory process; nervous system development; neurotoxicology; novel; promoter; response; sex; transcriptome

Regulation of transcription is central to proper nervous system development and functioning. Dysregulation of transcriptional and post-transcriptional regulatory pathways are associated with various neurodevelopmental diseases and disorders including developmental lead (Pb) exposure, which is associated with significant and persistent cognitive and behavioral deficits. Recent data from our lab have shown Pb-induced methylation changes at gene promoter regions and effects of Pb on various post-translational histone modifications (PTHMs), however, gene expression changes have not always corresponded to promoter methylation status and mismatches between PTHM levels and transcriptional regulation were also observed, suggesting that other factors may be influencing transcriptional changes in response to Pb exposures. Thus, mechanisms un- derlying altered transcriptional regulation in specific brain regions following Pb exposure cannot be completely explained by alterations in DNA methylation and PTHM levels. Preliminary data from our lab support the hy- pothesis that another mechanism, direct RNA methylation via N6-methyladenosine (m6A), may be an important post-transcriptional mechanism contributing to the functionally altered transcriptome after developmental Pb exposure. Like DNA, RNA is susceptible to dynamic, reversible chemical modification and this epitranscrip- tomic mechanism may play an important role in gene transcription and behavioral plasticity. This has not been investigated as a potential molecular mechanism involved in environmental neurotoxicology. The goal of this proposal is to test the novel hypothesis that the most abundant epitranscriptomic mark, m6A, plays an im- portant role in modulating RNA and influencing memory processes in male and female rats and, based on preliminary data, in regulating transcriptional and behavioral responses to developmental Pb exposure. The following aims will examine the contribution of m6A to memory processing in normal and Pb-exposed animals: Aim1: Examine transcriptome-wide profile of the m6A modification (using methylated RNA immunoprecipitation sequencing (MeRIP-seq)) in mPFC and hippocampus (CA1) in behaviorally naïve and behaviorally trained ani- mals of both sexes to assess its potential role as a dynamic epitranscriptomic mechanism involved in encoding and maintenance of memory, and, examine the extent to which the transcriptome-wide m6A enrichment profile is altered in animals with developmental Pb exposure; Aim2: Assess the functional significance of m6A in PFC and CA1 in memory processes in Pb-exposed animals using site-directed knockdown of the RNA demethylase gene, Fto, which will increase m6A levels in mPFC and CA1 and allow us to evaluate the effects of brain region -dependent reversal of decreased m6A levels on Pb-induced impaired memory processes. This work, the first to examine the dynamic modulation of RNA by m6A in PFC and CA1 in male and female normal and Pb- exposed rats, will result in new information regarding mRNA methylation as an effector mechanism for gene expression and behavioral plasticity in response to an environmental neurotoxicant.

转录的调节对于适当的神经系统发展和功能至关重要。失调的 转录和转录后调节途径与各种神经发育有关 疾病和疾病，包括发育铅（PB）暴露，这与重要和重要有关 持续的认知和行为定义。我们实验室的最新数据显示了PB诱导的甲基化 基因启动子区域的变化以及PB对各种翻译后Hisstone修饰的影响 （PTHM），但是，基因表达变化并不总是对应于启动子甲基化状态 还观察到PTHM水平和转录调节之间的不匹配，表明 其他因素可能会影响针对PB暴露的转录变化。那，机制不 PB暴露后特定大脑区域的转录调节变化改变不能完全是完全 解释了DNA甲基化和PTHM水平的改变。来自我们实验室的初步数据支持 通过N6-甲基读二氨酸（M6A）直接RNA甲基化的另一种机制可能是重要的 发育PB后功能变化的转录组的转录后机制 接触。像DNA一样，RNA易于动态，可逆的化学修饰，并且这种表观转录 - Tomic机制可能在基因转录和行为可塑性中起重要作用。这不是 被研究为参与环境神经毒理学的潜在分子机制。目标的目标 提案是检验新的假设，即最丰富的表演标记M6A扮演着一个不可思议的假设。 便携式在调节RNA和影响男性和雌性大鼠的记忆过程中的作用，并基于 初步数据，控制对发育PB暴露的转录和行为响应。这 以下目标将检查M6A对正常和PB暴露动物的记忆加工的贡献： AIM1：检查M6A修饰的整个转录组轮廓（使用甲基化RNA免疫沉淀 MPFC和海马（CA1）中的测序（Merip-Seq））在行为幼稚和行为训练的ani-中 两性的玛尔（Mal）评估其作为编码的动态表演机制的潜在作用 并维护记忆，并检查整个转录组M6A富集的程度 发育性PB暴露的动物会改变； AIM2：评估M6A在PFC中的功能意义 和CA1在PB暴露动物的记忆过程中使用RNA脱甲基酶的位定向敲低 基因，FTO，它将增加MPFC和CA1中的M6A水平，使我们能够评估大脑区域的影响 - 依赖性逆转在PB诱导的记忆过程中，M6A水平降低。这项工作，第一个 检查男性和女性正常和pb-中M6A对RNA的动态调节 暴露的大鼠将导致有关mRNA甲基化作为基因效应机制的新信息 响应环境神经毒性的表达和行为可塑性。