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

基本信息

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
项目类别：
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

项目摘要

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.

项目概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及其导致神经发育障碍（NDD）。 NDD 包括一系列高度普遍的病症在发育过程中表现出来，可能导致智力障碍、发育迟缓和自闭症频谱紊乱。最近的工作表明，许多染色质调节因子在 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，其相关具有主动转录功能。我们发现 H3K79me 在有丝分裂后高度丰富且动态调节神经元。我们的初步数据还表明 H3K79me 对于神经元功能至关重要。我们发现那个病人突变导致 Dot1L 甲基转移酶活性丧失，表明 H3K79me 的耗尽可导致 NDD。此外，我们发现 Dot1L 缺失会改变突触基因的转录并双向调节 GluA2，AMPA 受体亚基。最后，我们发现 Dot1L 条件敲除存在长期记忆缺陷（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 调节突触基因表达，部分 Dot1L 缺失会破坏这种调节，导致 NDD。在目标 1 中，我将使用杂合子定义由部分 Dot1L 丢失引起的染色质和转录破坏 Dot1L cKO 小鼠模型结合 H3K79me2 靶点切割和标记 (CUT&Tag) 以及 RNA测序。在目标 2 中，我将通过以下方式检查部分 Dot1L 丢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学研究实验。总之，这项工作将确定 Dot1L 在神经元功能和 NDD 等中的作用广泛地有助于理解染色质调节因子在大脑功能中的作用。