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.

项目概要/摘要 该提案的目的是确定染色质重塑剂 CHD7 的杂合突变如何 （染色质结构域解旋酶 DNA 结合蛋白 7）破坏新皮质的发育，导致 与 CHARGE 综合征相关的神经发育异常。 CHARGE 综合征的特点是 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 （自闭症谱系障碍）。拟议的研究小组最近发现了 Chd7 杂合子的新皮质异常 小鼠，表明 CHD7 参与关键的、时间上不同的发育过程。初步数据表明 这些小鼠的新皮质的前后 (​​A-P) 模式被破坏，CHD7 直接调节 A-P 模式的主调节因子 Nr2f1 (COUP-TF1) 的表达。这些小鼠还表现出皮质 发育不全，暗示 Chd7 参与皮质生长。此外，深层的兴奋性和抑制性突触 前额皮质 (PFC) 的主要神经元受到 Chd7 单倍体不足的影响。双方都是执行官 功能障碍和 ASD 与 PFC 功能障碍有关。因此，这些发现提供了人们长期寻求的 有机会确定 CHD7 单倍体不足扰乱新皮质发育的机制。 Chd7 功能是上下文相关的；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。该项目的具体目标是 1) 检验 Chd7 单倍体不足的假设 破坏新皮质的 A-P 模式，2) 定义 Chd7 在神经中的分子和细胞功能 新皮质和皮层下层的干/祖细胞，3) 检验 Chd7 具有细胞类型特异性的假设 PFC 中突触发育的功能。该团队将使用标准和创新的方法 可视化和量化 A-P 模式标记的表达模式和水平，并量化区域、细胞 以及兴奋性和抑制性神经发生的性别特异性异常。下一代散装和单细胞 测序方法将用于识别不同区域的转录和染色质变化 两性 Chd7 缺陷的发育中新皮质的细胞类型。多机构团队汇聚一堂 CHD 蛋白的神经发育功能（Basson）、转化和建模研究方面的专业知识 CHARGE 综合征 (Martin)、皮质发育基因组学 (Kwan) 和突触生理学 (Andreae)。 总之，这项工作将全面了解 CHD7 单倍体不足对 新皮质发育的多个过程并识别神经中 CHD7 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 识别神经发育机制和回路所需的知识和工具 与 CHARGE 综合征和其他相关疾病相关的特定行为和认知表型。