The Role of H3K79 Methylation and Dot1L in Neuronal Function and Neurodevelopmental Disorders

H3K79 甲基化和 Dot1L 在神经元功能和神经发育障碍中的作用

• 批准号: 10750689
• 负责人: Marissa Maroni
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750689
• 关键词: AMPA Receptors; Address; Affect; Behavior; Behavioral; Brain; Cell Nucleus; Cells; Cerebellum; Child; Chromatin; Cognition; Coupled; DNA; Data; Deposition; Development; Developmental Delay Disorders; Electrophysiology (science); Gene Expression; Genes; Genetic Transcription; Genomic Segment; Heterozygote; Hippocampus; Histone-Lysine N-Methyltransferase; Histones; Individual; Intellectual functioning disability; Knockout Mice; Label; Language Development; Lead; Loss of Heterozygosity; Lysine; Measures; Memory impairment; Methylation; Methyltransferase; Modeling; Mus; Mutate; Mutation; Neurodevelopmental Disorder; Neurons; Output; Patients; Phenotype; Play; Post-Translational Protein Processing; Proliferating; Prosencephalon; Protein Analysis; Proteins; Regulation; Reversal Learning; Role; Running; Synapses; Synaptic Receptors; Time; Tissues; Transcriptional Regulation; Ultrasonics; Western Blotting; Work; autism spectrum disorder; behavior test; causal variant; cell type; conditional knockout; de novo mutation; epigenetic regulation; exome sequencing; experimental study; histone methylation; histone methyltransferase; insight; knock-down; long term memory; loss of function; loss of function mutation; mouse model; nerve stem cell; neuron development; object recognition; patch clamp; postmitotic; protein function; receptor; transcriptome sequencing; vocalization; wound

PROJECT SUMMARY This proposal aims to identify the role of the histone lysine methyltransferase Dot1L in neuronal function and its contribution to neurodevelopmental disorders (NDDs). NDDs include a spectrum of highly prevalent conditions that manifest during development that can cause intellectual disability, developmental delays, and autism spectrum disorder. Recent work demonstrated that many chromatin regulators are mutated in NDDs, including the histone methyltransferase Dot1L. Dot1L methylates histone 3 of lysine 79 (H3K79me) which is associated with active transcription. We found that H3K79me is highly abundant and dynamically regulated in postmitotic neurons. Our preliminary data also indicate that H3K79me is critical for neuronal function. We found that patient mutations result in a loss of Dot1L methyltransferase activity indicating that depletion of H3K79me can cause NDDs. Further, we found that Dot1L depletion alters transcription of synaptic genes and bidirectionally regulates GluA2, an AMPA receptor subunit. Finally, we found long-term memory deficits in Dot1L conditional knockout (cKO) mice. However, the role of Dot1L in neuronal function and cognition remain unclear. I hypothesize that Dot1L regulates synaptic gene expression and that partial Dot1L loss disrupts this regulation leading to NDDs. In Aim 1, I will define chromatin and transcriptional disruptions caused by partial Dot1L loss using a heterozygous Dot1L cKO mouse model coupled with H3K79me2 cleavage under targets and tagmentation (CUT&Tag) and RNA-sequencing. In Aim 2, I will examine the impact of partial Dot1L loss on neuronal function and cognition by using the heterozygous Dot1L cKO mouse model and controls to perform electrophysiology and behavioral experiments. Cumulatively, this work will establish a role for Dot1L in neuronal function and NDDs and more broadly will contribute to understanding of the role of chromatin regulators in brain function.

项目摘要 该提案旨在确定组蛋白赖氨酸甲基转移酶Dot1L在神经元功能中的作用， 神经发育障碍（NDD）。NDD包括一系列高度流行的疾病 在发育过程中表现出来，可能导致智力残疾，发育迟缓和自闭症 谱系障碍最近的研究表明，许多染色质调节基因在NDD中发生突变，包括 组蛋白甲基转移酶Dot1L。Dot1L甲基化赖氨酸79（H3K79me）的组蛋白3， 活跃的转录。我们发现H3K79me在有丝分裂后细胞中是高度丰富的，并且是动态调节的。 神经元我们的初步数据还表明H3K79me对神经元功能至关重要。我们发现那个病人 突变导致Dot1L甲基转移酶活性的丧失，表明H3K79me的缺失可导致 NDD。此外，我们发现Dot1L缺失改变了突触基因的转录，并双向调节 GluA2，AMPA受体亚单位。最后，我们发现Dot1L条件性基因敲除的小鼠存在长时程记忆缺陷 (cKO)小鼠然而，Dot1L在神经元功能和认知中的作用仍不清楚。我假设 Dot1L调节突触基因的表达，部分Dot1L的缺失破坏了这种调节，导致NDD。 在目标1中，我将使用杂合的Dot1L缺失来定义由Dot1L部分缺失引起的染色质和转录破坏。 Dot1L cKO小鼠模型与靶下的H3K79me2切割和标签化（CUT&Tag）偶联， RNA测序。在目标2中，我将通过以下方法研究部分Dot1L缺失对神经元功能和认知的影响： 使用杂合Dot1L cKO小鼠模型和对照进行电生理学和行为 实验累积起来，这项工作将建立Dot1L在神经元功能和NDD等方面的作用 广泛地将有助于理解染色质调节剂在脑功能中的作用。