Dissecting Cell Type Specific Functions of CHD7 in Development of the Neocortex

剖析 CHD7 在新皮质发育中的细胞类型特异性功能

• 批准号: 10732823
• 负责人: Laura Andreae
• 金额: $ 56.21万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10732823
• 关键词: ATAC-seq; Acute; Affect; Animal Model; Anterior; Behavioral; Brain; Brain Diseases; CHARGE syndrome; CHD7 gene; Cell Differentiation process; Cell physiology; Cells; Cellular biology; ChIP-seq; Chromatin; Chromatin Structure; Cognitive; Confocal Microscopy; Data; Development; Developmental Process; Disease; Electrophysiology (science); Embryo; Enhancers; Excitatory Synapse; Executive Dysfunction; Exhibits; Functional disorder; Gene Deletion; Gene Expression; Gene Mutation; Genes; Genetic; Genetic Transcription; Genomics; Growth; Heterozygote; Human; In Situ Hybridization; Inhibitory Synapse; Knowledge; Link; Methods; Molecular; Mus; Mutate; Mutation; Neocortex; Neurodevelopmental Disorder; Neurons; Pathogenicity; Pattern; Phenotype; Physiologic pulse; Physiology; Population; Prefrontal Cortex; Process; Proliferating; Proteins; Regulatory Element; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Slice; Study models; Synapses; Syndrome; Testing; Time; Variant; Visual; Visual Cortex; Visualization; Wild Type Mouse; Work; area striata; autism spectrum disorder; cell type; chromatin remodeling; developmental disease; digital; excitatory neuron; experimental study; individuals with autism spectrum disorder; inhibitory neuron; innovation; multiple omics; neocortical; nerve stem cell; neurogenesis; neuropsychiatric disorder; next generation sequencing; overexpression; patch clamp; postnatal; progenitor; sex; stem cell proliferation; synaptogenesis; targeted treatment; tool; translational model

PROJECT SUMMARY/ABSTRACT The aim of this proposal is to determine how heterozygous mutation of the chromatin remodeler CHD7 (Chromodomain Helicase DNA-binding protein 7) disrupts development of the neocortex, leading to the neurodevelopmental anomalies associated with CHARGE syndrome. CHARGE syndrome is characterized by substantial behavioral and cognitive problems, including executive dysfunction and Autism Spectrum Disorder (ASD). The proposed research team recently identified abnormalities in the neocortex of Chd7 heterozygous mice, implicating CHD7 in key, temporally distinct developmental processes. Preliminary data suggest that anterior-posterior (A-P) patterning of the neocortex is disrupted in these mice and that CHD7 directly regulates the expression of a master regulator of A-P patterning, Nr2f1 (COUP-TF1). These mice also exhibit cortical hypoplasia, implicating Chd7 in cortical growth. Furthermore, excitatory and inhibitory synapses onto deep layer principal neurons of the prefrontal cortex (PFC) are affected by Chd7 haploinsufficiency. Both executive dysfunction and ASD have been linked to PFC dysfunction. Thus, these findings provide the long-sought opportunity to identify the mechanisms whereby CHD7 haploinsufficiency disrupts neocortical development. Chd7 function is context-dependent; hence, a significant focus will be to define sex-, region- and cell-type-specific functions and mechanisms. This project's specific aims are to 1) test the hypothesis that Chd7 haploinsufficiency disrupts A-P patterning of the neocortex, 2) define the molecular and cellular functions of Chd7 in neural stem/progenitor cells of the neocortex and subpallium and 3) test the hypothesis that Chd7 has cell-type-specific functions in the development of synapses in the PFC. The team will use standard and innovative methods to visualize and quantify the expression patterns and levels of markers of A-P patterning, and quantify region-, cell- and sex-specific abnormalities in excitatory and inhibitory neurogenesis. Bulk and single cell next generation sequencing approaches will be used to identify transcriptional and chromatin changes in different regions and cell types of the Chd7-deficient developing neocortex in both sexes. The multi-institute team brings together expertise in neurodevelopmental functions of CHD proteins (Basson), translational and modeling studies of CHARGE syndrome (Martin), genomics of cortical development (Kwan), and synapse physiology (Andreae). Together, this work will provide a comprehensive understanding of the impacts of CHD7 haploinsufficiency on multiple processes in development of the neocortex and identify cell-type-specific functions for CHD7 in neural progenitor differentiation and PFC circuit assembly. Successful completion of this work will generate the knowledge and tools necessary to identify the neurodevelopmental mechanisms and circuits that underlie specific behavioral and cognitive phenotypes associated with CHARGE syndrome and other related disorders.

项目总结/摘要 本研究的目的是确定染色质重塑基因CHD 7的杂合突变如何影响 （染色体结构域解旋酶DNA结合蛋白7）破坏新皮层的发育，导致 与CHARGE综合征相关的神经发育异常。CHARGE综合征的特征是 严重的行为和认知问题，包括执行功能障碍和自闭症谱系障碍 （ASD）中指定的值。拟议的研究小组最近发现了Chd 7杂合子的新皮层异常， 小鼠，暗示CHD 7在关键的，时间上不同的发育过程。初步数据显示， 在这些小鼠中，新皮层的前-后（A-P）模式被破坏，并且CHD 7直接调节 A-P模式的主要调节因子Nr 2f 1（COUP-TF 1）的表达。这些小鼠也表现出皮质 发育不全，暗示皮质生长中的Chd 7。此外，深层的兴奋性和抑制性突触 前额叶皮层（PFC）的主要神经元受到Chd 7单倍不足的影响。均为执行 功能障碍和ASD与PFC功能障碍有关。因此，这些发现提供了长期寻求的 有机会确定CHD 7单倍不足破坏新皮质发育的机制。 Chd 7的功能依赖于环境，因此，一个重要的重点将是定义性别，区域和细胞类型特异性 功能和机制。本项目的具体目标是：1）检验Chd 7单倍不足的假设， 破坏了新皮层的A-P模式，2）定义了Chd 7在神经元中的分子和细胞功能， 新皮层和下腭的干/祖细胞和3）测试Chd 7具有细胞类型特异性的假设。 在PFC突触发育中的作用。该团队将使用标准和创新的方法， 可视化和定量A-P模式标记物的表达模式和水平，并定量区域，细胞， 以及兴奋性和抑制性神经发生的性别特异性异常。下一代散装和单电池 测序方法将用于鉴定不同区域的转录和染色质变化， 两种性别中Chd 7缺陷发育中的新皮质的细胞类型。多机构团队汇集了 CHD蛋白质的神经发育功能的专业知识（Basson）， CHARGE综合征（Martin）、皮质发育的基因组学（Kwan）和突触生理学（André）。 总之，这项工作将提供一个全面的理解的影响，CHD 7单倍不足， 新皮层发育中的多个过程，并确定神经元中CHD 7的细胞类型特异性功能。 祖细胞分化和PFC电路组装。成功完成这项工作将产生 知识和必要的工具，以确定神经发育机制和电路的基础 与CHARGE综合征和其他相关疾病相关的特定行为和认知表型。