The role of H3K36 methyltransferases on non-CpG methylation patterning in the mammalian brain

H3K36 甲基转移酶对哺乳动物大脑非 CpG 甲基化模式的作用

• 批准号: 10764214
• 负责人: Nicole Hamagami-Samson
• 金额: $ 3.36万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10764214
• 关键词: ASH1L gene; Adult; Affect; Affinity; Binding; Brain; Child; Chromatin; Clinical; DNA Methylation; DNA Modification Methylases; DNA Modification Process; Deposition; Development; Dinucleoside Phosphates; Disease; Enhancers; Enzymes; Epigenetic Process; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic study; Genome; Genomic approach; Genomics; Goals; Histones; Human Genetics; Impairment; In Vitro; Knockout Mice; Knowledge; Laboratories; Link; Mammalian Cell; Measures; Mediating; Methods; Methyl-CpG-Binding Protein 2; Methylation; Methyltransferase; Modeling; Modification; Molecular; Mutant Strains Mice; Mutate; Mutation; Nervous System; Neurodevelopmental Disorder; Neuroglia; Neurologist; Neurons; Outcome; PWWP Domain; Pathology; Pathway interactions; Pattern; Play; Process; Publishing; Reading; Regulation; Research; Research Training; Role; Site; Syndrome; System; Testing; Training; Transcriptional Regulation; Up-Regulation; Wild Type Mouse; Work; Writing; autism spectrum disorder; brain dysfunction; career; chromatin modification; conditional knockout; epigenomics; gene regulatory network; genome-wide; genome-wide analysis; genomic data; histone methyltransferase; histone modification; in vivo; insight; knock-down; methylation pattern; neurodevelopment; neuron development; novel; programs; recruit; transcriptome sequencing

Project Summary: Human genetic studies have linked autism and related neurodevelopmental disorders (NDD) to disruption of genes encoding epigenetic factors. While mutations in these genes can affect more than one pathway that leads to ASD, identification of a common molecular pathway across genetic causes of ASD can provide insight into broadly applicable therapies. Recent evidence from our laboratory suggests that a neuronal-specific form of DNA methylation is a shared epigenetic modification that is disrupted in ASD-associated neurodevelopmental disease. Though DNA methylation is classically considered to only occur in mammalian cells in the CpG context, neurons are uniquely enriched for methylation in non-CpG contexts established by DNMT3A. This non-CpG methylation primarily occurs at CA dinucleotides (mCA) and is critical for proper neuronal development and function. Though mCA plays an important role in regulating neuronal gene expression, it is not known how the mCA landscape is faithfully established across the neuronal genome, and whether additional NDDs involve disruption of mCA. Recent studies outside the nervous system have suggested that histone modifications may play a central role in directing DNMT3A-mediated DNA methylation across the genome, but it is not known to how these histone modifications influence DNMT3A and mCA throughout the neuronal genome. Intriguingly, mutations in H3K36 histone methyltransferases have been recently identified in ASD gene studies. Additionally, recent studies conducted outside the nervous system have suggested that H3K36 methylation recruits DNMT3A and regulates its activity. In this proposal, I will determine how disruption of H3K36 methylation-mediated interactions with DNMT3A disturbs critical patterns of mCA across the neuronal genome to drive brain dysfunction. In Aim 1, I will disrupt NDD-relevant H3K36 methyltransferases and measure how DNMT3A recruitment and mCA are affected. This will define the mechanisms underlying histone and DNA modification dynamics on regulating neuronal transcription and building our basic understanding of how disruption of mCA drives disease. In Aim 2, I will explore how loss of H3K36 dimethylase NSD1 causes dysregulation of neuronal genes via shared neuronal chromatin pathology observed across heterogenous clinical syndromes of ASD. I will use genomic approaches to examine how altered DNA methylation as a result of NSD1 loss affects enhancer activity to drive transcriptional dysregulation in nervous system dysfunction and disease. This analysis will begin to identify key downstream chromatin associated factors across a common molecular pathway involved in multiple genetic causes of ASD to provide insight into functional cellular outcomes and broadly applicable therapies.

项目概要： 人类遗传学研究已将自闭症和相关神经发育障碍 (NDD) 与智力障碍联系起来。 编码表观遗传因子的基因。虽然这些基因的突变可以影响不止一种途径，导致 对于自闭症谱系障碍（ASD），识别自闭症谱系障碍遗传原因的共同分子途径可以提供深入了解 广泛适用的疗法。我们实验室的最新证据表明，神经元特异性形式的 DNA 甲基化是一种共有的表观遗传修饰，在自闭症谱系障碍相关的神经发育疾病中被破坏。 尽管 DNA 甲基化传统上被认为仅发生在哺乳动物细胞的 CpG 环境中， 神经元在 DNMT3A 建立的非 CpG 环境中独特地富集甲基化。这种非 CpG 甲基化主要发生在 CA 二核苷酸 (mCA)，对于神经元的正常发育和发育至关重要 功能。尽管 mCA 在调节神经元基因表达中发挥着重要作用，但尚不清楚其如何发挥作用。 mCA 景观在整个神经元基因组中忠实地建立，并且额外的 NDD 是否涉及 mCA 的破坏。最近神经系统之外的研究表明组蛋白修饰可能 在引导 DNMT3A 介导的 DNA 甲基化跨基因组方面发挥核心作用，但尚不清楚 这些组蛋白修饰如何影响整个神经元基因组中的 DNMT3A 和 mCA。 有趣的是，最近在 ASD 基因中发现了 H3K36 组蛋白甲基转移酶的突变 研究。此外，最近在神经系统之外进行的研究表明，H3K36 甲基化招募 DNMT3A 并调节其活性。在这个提案中，我将确定如何中断 H3K36 甲基化介导的与 DNMT3A 的相互作用扰乱了神经元基因组中 mCA 的关键模式 来驱动大脑功能障碍。在目标 1 中，我将破坏 NDD 相关的 H3K36 甲基转移酶并测量如何 DNMT3A 募集和 mCA 受到影响。这将定义组蛋白和 DNA 的潜在机制 调节神经元转录的修饰动力学并建立我们对如何调节的基本理解 mCA 的破坏会导致疾病。在目标 2 中，我将探讨 H3K36 二甲基酶 NSD1 的缺失如何导致 通过在异质临床中观察到的共享神经元染色质病理学导致神经元基因失调 ASD 综合症。我将使用基因组方法来检查 NSD1 如何改变 DNA 甲基化 丢失会影响增强子活性，从而驱动神经系统功能障碍和疾病中的转录失调。 该分析将开始识别常见分子中关键的下游染色质相关因素 涉及 ASD 多种遗传原因的通路，可深入了解功能性细胞结果和 广泛适用的疗法。

项目成果

NSD1 deposits histone H3 lysine 36 dimethylation to pattern non-CG DNA methylation in neurons.

• DOI: 10.1016/j.molcel.2023.04.001
• 发表时间: 2023-04
• 期刊: Molecular cell
• 影响因子: 16
• 作者: Nicole Hamagami;Dennis Y. Wu;Adam W. Clemens;Sabin A. Nettles;Aidan Li;Harrison W. Gabel