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对长期结果的影响

• 批准号: 10658020
• 负责人: JAY S SCHNEIDER
• 金额: $ 51.54万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10658020
• 关键词: Address; Adenosine; Adverse effects; Affect; Animal Model; Animals; Antibodies; Auditory; Basic Science; Behavior; Behavioral; Brain; Brain region; Chemical Dynamics; Child; Cognitive; Cognitive deficits; Cues; DNA; DNA Methylation; Data; Development; Dimensions; Environment; Environmental Exposure; Environmental Impact; Female; Functional disorder; Gene Expression; Genes; Genetic Transcription; Hippocampus; Immunoprecipitation; Impaired cognition; Lead; Learning; Life; Link; Literature; Medial; Mediating; Memory; Memory impairment; Messenger RNA; Methylation; Modification; Molecular; Mus; Neurodevelopmental Disorder; Neuronal Plasticity; Neurotoxins; Outcome; Pathologic; Pathway interactions; Performance; Play; Post-Transcriptional Regulation; Predisposition; Prefrontal Cortex; Process; Promoter Regions; Public Health; Publishing; RNA; RNA methylation; Rattus; Regulatory Pathway; Reporting; Role; Testing; Time; Training; Transcript; Transcriptional Regulation; Translations; Untranslated RNA; Viral; Work; adverse outcome; behavioral outcome; behavioral plasticity; behavioral response; conditioned fear; crosslink; design; epitranscriptome; epitranscriptomics; fat mass and obesity-associated protein; gene environment interaction; histone modification; improved; knock-down; lead exposure; male; memory consolidation; memory process; memory recall; nervous system development; neurotoxicity; novel; posttranscriptional; remediation; response; sex; stressor; transcriptome; transcriptomics; virtual

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 often results in significant and persistent cognitive and behavioral deficits. We have previously reported Pb-induced methylation changes at gene promoter regions and effects of Pb on various post-translational histone modifications (PTHMs) in hippo- campus (HIPP) and medial prefrontal cortex (mPFC) in animals with Pb-induced cognitive dysfunction, with sex, amount of Pb exposure, and developmental window of exposure as effect modifiers. While prior work has focused on effects of Pb on DNA-related mechanisms, an additional molecular transcriptional control mechanism that has not been studied in this regard is RNA modification. Preliminary data from our lab support the hypothesis that direct RNA methylation via N6-methyladenosine (m6A/m), may be an important mechanism contributing to the functionally altered transcriptome after developmental Pb exposure. Direct modifications of RNA appear to provide an additional layer of control over RNA function for the fine-tuning of transcriptomic responses to the environment, critical for normal plasticity and memory function. Considering the negative impact that Pb exposure has on neural plasticity and memory, we suggest that there is a potentially important role of m6A/m in post-transcriptional regulation of gene-specific responses associated with Pb-induced plasticity/ memory impairments. Our hypothesis is that this abundant epitranscriptomic mark, m6A/m, not only plays an important role in modulating memory processes under normal circumstances but is altered by early Pb exposure with an adverse influence on transcriptional and behavioral responses to Pb. We will test this hypothesis in the following aims: Aim1: Examine the extent to which transcriptome-wide m6A/m profiles in mPFC and HIPP CA1 and are altered by developmental Pb exposure in males and females and associated with expression of memory deficits. Hypothesis: There will be brain region-distinct transcriptome-wide enrichment profiles of m6A/m (detected by m6A-eCLIP (enhanced UV-crosslinking and immunoprecipitation), particularly associated with memory/plasticity-related genes and pathways, that are aberrantly modulated in Pb-exposed, memory-impaired animals; Aim2: Examine the functional significance of m6A/m modification in mPFC and CA1 in Pb-exposed animals through manipulation of m6A/m levels. As preliminary data indicate decreased m6A levels and increased FTO expression in Pb-exposed rats with memory deficits, we will explore the potential efficacy of viral-mediated targeted knockdown of FTO on Pb-induced memory impairments and m6A/m levels and memory/plasticity-related gene expression in mPFC and CA1. We hypothesize that by increasing m6A/m levels we will at least partially ameliorate the Pb-induced memory impairment.

转录调节对于神经系统的正常发育和功能至关重要。失调 转录和转录后调节途径与各种神经发育相关 疾病和失调，包括发育性铅 (Pb) 暴露，这通常会导致严重和严重的后果 持续的认知和行为缺陷。我们之前曾报道过 Pb 诱导的甲基化变化 河马基因启动子区域和 Pb 对各种翻译后组蛋白修饰 (PTHM) 的影响 铅诱发认知功能障碍动物的大脑皮质 (HIPP) 和内侧前额叶皮层 (mPFC) 性别、铅暴露量和发育暴露窗口作为影响调节剂。虽然之前的工作有 重点关注 Pb 对 DNA 相关机制的影响，这是一种额外的分子转录控制 这方面尚未研究的机制是RNA修饰。来自我们实验室支持的初步数据 通过 N6-甲基腺苷 (m6A/m) 直接进行 RNA 甲基化的假设可能是一种重要机制 导致发育铅暴露后转录组功能发生改变。直接修改 RNA 似乎为 RNA 功能提供了额外的控制层，用于转录组的微调 对环境的反应，对于正常的可塑性和记忆功能至关重要。考虑到负面影响 铅暴露对神经可塑性和记忆力的影响，我们认为有一个潜在的重要影响 m6A/m 在与 Pb 诱导相关的基因特异性反应的转录后调节中的作用 可塑性/记忆障碍。我们的假设是，这种丰富的表观转录组标记 m6A/m 不仅 在正常情况下，它在调节记忆过程中起着重要作用，但会被早期改变 铅暴露会对铅的转录和行为反应产生不利影响。我们将测试这个 假设的目标如下： 目标 1：检查转录组范围内 m6A/m 分布的程度 mPFC 和 HIPP CA1 会因男性和女性发育过程中的铅暴露以及相关的变化而改变 并伴有记忆缺陷的表现。假设：整个大脑区域都会存在不同的转录组 m6A/m 的富集曲线（通过 m6A-eCLIP（增强型 UV 交联和免疫沉淀）检测）， 特别与记忆/可塑性相关的基因和途径相关，这些基因和途径在 接触铅、记忆受损的动物；目标 2：检查 m6A/m 修饰的功能意义 通过操纵 m6A/m 水平来观察铅暴露动物的 mPFC 和 CA1。初步数据表明 在患有记忆缺陷的铅暴露大鼠中，m6A 水平降低，FTO 表达增加，我们将探讨 病毒介导的 FTO 靶向敲低对 Pb 诱导的记忆障碍的潜在功效 mPFC 和 CA1 中的 m6A/m 水平和记忆/可塑性相关基因表达。我们假设通过 增加 m6A/m 水平，我们将至少部分改善 Pb 引起的记忆障碍。