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.

转录调控是神经系统正常发育和功能的核心。失调 转录和转录后调节途径与各种神经发育相关， 疾病和紊乱，包括发育期铅（Pb）暴露， 持续的认知和行为缺陷。我们实验室的最新数据显示， 基因启动子区的变化和铅对各种翻译后组蛋白修饰的影响 然而，基因表达变化并不总是与启动子甲基化状态相对应 PTHM水平和转录调控之间的错配也被观察到，这表明， 其他因素可能会影响转录变化响应铅暴露。因此，机制不- 铅暴露后，在特定的脑区域中发生转录调节的改变， 这可以通过DNA甲基化和PTHM水平的改变来解释。我们实验室的初步数据支持- 假设另一种机制，通过N6-甲基腺苷（m6 A）直接RNA甲基化，可能是一个重要的机制。 转录后机制有助于发育铅后转录组功能改变 exposure.像DNA一样，RNA对动态的、可逆的化学修饰敏感，这种表观转录- 体细胞机制可能在基因转录和行为可塑性中起重要作用。这还没有得到 研究作为一个潜在的分子机制参与环境神经毒理学。这个目标 建议是测试新的假设，即最丰富的epitranscriptomic标记，m6 A，发挥免疫调节作用。 在调节RNA和影响雄性和雌性大鼠的记忆过程中起重要作用， 初步数据，在调节转录和行为反应发育铅暴露。的 以下目标将检查m6 A对正常和铅暴露动物记忆处理的贡献： 目的1：检查m6 A修饰的全转录组概况（使用甲基化RNA免疫沉淀 MeRIP-seq））在mPFC和海马（CA 1）的行为幼稚和行为训练的动物。 评估其作为参与编码的动态表观转录组机制的潜在作用 和记忆的维持，并检查转录组范围内m6 A富集概况的程度， 在发育期铅暴露的动物中改变;目的2：评估m6 A在PFC中的功能意义 利用RNA去甲基化酶的定点敲低研究铅暴露动物记忆过程中的CA 1和CA 2的表达 Fto基因，这将增加mPFC和CA 1中的m6 A水平，并使我们能够评估大脑区域的影响。 铅诱导的受损记忆过程中m6 A水平降低的依赖性逆转。这项工作，第一 检测正常男性和女性以及铅中毒患者PFC和CA 1中m6 A对RNA的动态调节。 暴露的大鼠，将导致新的信息，mRNA甲基化作为基因的效应机制， 表达和行为可塑性的反应环境神经毒物。