Genetic Studies of Cortex Structure and Development

皮层结构和发育的遗传学研究

• 批准号: 9360012
• 负责人: JOHN L. R. RUBENSTEIN
• 金额: $ 63.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9360012
• 关键词: Autistic Disorder; Automobile Driving; Binding; Binding Proteins; Biological Assay; CRISPR/Cas technology; Cell Separation; Cell physiology; Cells; Cerebral Palsy; Cerebral cortex; Code; DNA sequencing; Development; Embryo; Enhancers; Epilepsy; Fibroblast Growth Factor; Foundations; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genetic study; Histones; Human; Human Genetics; Mental Retardation; Mental deficiency; Mitotic; Molecular; Mutation; Nature; Neurons; Pathway interactions; Pattern; Phenotype; Regulator Genes; Regulatory Element; Research; Role; Schizophrenia; Signal Transduction; Structure; Testing; Transcriptional Regulation; Transgenic Mice; Translating; Ventricular; Work; chromatin immunoprecipitation; developmental disease; emx2 protein; epigenomics; experimental study; genetic information; genetic variant; in vivo; insight; loss of function; loss of function mutation; mutant; neuropsychiatric disorder; progenitor; promoter; public health relevance; response; subventricular zone; transcription factor; transcriptome sequencing

PROJECT SUMMARY Dysregulation of the cerebral cortex is central to human developmental disorders such as epilepsy, mental deficiency, autism and schizophrenia. During development, cortical progenitors generate the projection neurons of the different cortical subdivisions. Understanding the genetic circuitry controlling the development and function of these neurons provides an essential foundation for interpreting human allele variants that are enriched in people who have neuropsychiatric disorders. To elucidate this genetic circuitry, we must define the transcription factors (TF), and regulatory elements and of the coding regions that they control. The proposed research, which concentrates on cortical regionalization, involves the systematic identification of TFs, and the regulatory elements and genes downstream of TFs. Currently, the regional-specification functions of a few TFs in embryonic cortical progenitors are known, and little is known about their direct transcription targets, the nature of the regulatory elements that these TFs control, and the transcriptional circuitry that integrates development and function of these cells. Here we propose to make inroads into each of these components of the TF hierarchy regulating cortical development. Furthermore, we aim to elucidate transcriptional mechanisms through which patterning of cortical progenitors is transmitted to, and maintained in, cortical neurons. We hypothesize that enhancers active in the ventricular zone, subventricular zone and the cortical plate are differentially bound by TFs that drive expression of region/layer-specific genes in post-mitotic cortical neurons. The enhancers serve as protein-binding modules that translate rostrocaudal gradients of TFs in cortical progenitors into region-specific expression in cortical neurons. Herein we focus on the transcriptional mechanisms controlling the generation of different regions of the cerebral cortex (cortical regionalization). The Five Specific Aims extend upon our earlier work on regionalization of cortical progenitors by FGF-signaling, TFs and enhancer elements. Here we investigate transcriptional regulation of cortical patterning by defining the TFs, and other genes, that are regulated by COUPTF1, EMX2, and PAX6 (Aim 1). We then use chromatin immunoprecipitation-DNA sequencing (ChIP-Seq) to define the regulatory element (RE) and gene targets of COUPTF1, EMX2, and PAX6 (Aim 2). Next, we use fluorescent activated cell sorting (FACS) to purify cells from the VZ, SVZ, CP and layers 5&6 to elucidate the epigenomic states of REs (and genes) using Histone ChIP-Seq. This will help us understand the molecular mechanisms that transmit regional patterning information from cortical progenitors to neurons (Aim 3). Finally, we define the function of REs related to cortical patterning using transgenic mice to assess RE activity (Aims 4) and REs deletions to define their role in gene regulation (Aim 5). Once integrated with human genetic information, this will enable us to gain powerful insights into how abnormalities in specific gene networks cause human neuropsychiatric disorders.

项目摘要 大脑皮层的失调是人类发育障碍的核心，例如癫痫，心理 缺乏症，自闭症和精神分裂症。在开发过程中，皮质祖细胞产生投影 不同皮质细分的神经元。了解控制发展的遗传回路 这些神经元的功能为解释人类等位基因变体提供了重要基础 丰富了患有神经精神疾病的人。为了阐明这种遗传回路，我们必须定义 转录因子（TF），监管元素以及它们控制的编码区域。提议 研究集中于皮质区域化的研究涉及TFS的系统鉴定，以及 TFS下游的调节元素和基因。目前，几个TF的区域规范功能 在胚胎皮质祖细胞中，知之甚少，对它们的直接转录靶标的知之甚少 这些TFS控制的调节元素的性质以及集成的转录电路 这些细胞的开发和功能。在这里，我们建议将这些组成部分侵入 调节皮质发展的TF层次结构。此外，我们旨在阐明转录机制 皮质祖细胞的模式传递到皮质神经元并将其维持。我们 假设在心室区域，室内区和皮质板中活跃的增强剂是 通过TF的差异结合，这些TF驱动有丝分裂后皮质神经元中区域/层特异性基因的表达。 增强剂用作蛋白质结合模块，可翻译皮质中TF的tfs梯度 祖细胞在皮质神经元中特异性表达。在此，我们专注于转录 控制大脑皮层不同区域的产生的机制（皮质区域化）。这 五个特定的目标扩展了我们较早通过FGF信号来进行皮质祖细胞区域化的工作， TFS和增强子元素。在这里，我们通过定义了皮质图案的转录调控 由couptf1，emx2和pax6调节的TF和其他基因（AIM 1）。然后我们使用染色质 免疫沉淀-DNA测序（CHIP-SEQ）定义调节元件（RE）和基因靶标 COUPTF1，EMX2和PAX6（AIM 2）。接下来，我们使用荧光激活的细胞分选（FACS）净化细胞 从VZ，SVZ，CP和层5和6，使用组蛋白阐明RES（和基因）的表观基因组状态 chip-seq。这将有助于我们了解传输区域模式信息的分子机制 从皮质祖细胞到神经元（AIM 3）。最后，我们定义与皮质图案相关的RES的功能 使用转基因小鼠评估RE活性（目标4）并进行删除以定义其在基因调节中的作用 （目标5）。一旦与人类遗传信息融为一体，这将使我们能够对 特定基因网络的异常引起人类神经精神疾病。