Genetic Studies of Cortex Structure and Development

项目概要（30行） 皮质发育失调是癫痫、智力缺陷、自闭症和精神分裂症的核心。 皮质祖细胞产生大脑皮质和海马的投射神经元。理解 控制大脑皮层发育和功能的遗传电路对于解释人类等位基因是必不可少的。 神经精神疾病患者的基因变异为了阐明基因电路驱动 皮质祖细胞和神经元的发育，我们必须定义必需的转录因子（TF）， 调控元件（RE）和编码区。皮质区域化和层状研究的建议 规范，将克服这些障碍，通过识别和功能特性的RE 与皮质发育有关此外，染色质修饰剂对RE的调节对于它们的功能至关重要。 适当的时空活动8，9.在这里，我们建议对这些主题进行研究。我们的目标是 阐明转录机制，通过该机制，皮质祖细胞的图案被传递到， 维持在皮层神经元中。 在这个提议中，我们将研究转录因子和它们的靶点（假定的RE或pRE）调节皮层模式 和层压（目标1）。我们将使用纯化的pREs的区域和层状表观基因组状态进行分析。 皮质祖细胞和分化中的神经元（目的2和3）。我们将定义pRE的活性和功能 使用转基因小鼠和增强子缺失（Aim 4）。最后，为了确定染色质修饰剂在 皮层区域化和分层，我们将研究一个染色质修饰条件突变体（Kdm 6 b cKO，Aim 5）。 这些研究将我们的研究从单个基因的功能转向了解如何 转录网络协调皮质发育。我们将整合，我们的合作大规模 用我们的ChIP-Seq（染色质免疫沉淀-测序）对前脑pRES 1，43，44进行基因组学分析 分析，以确定区域和层特异性皮质pRE。这些信息可以导致关于 非编码序列中的人类疾病等位基因。此外，我们的研究结果将提供关键信息， 皮质祖细胞和神经元的遗传控制。一旦与人类遗传信息整合， 将导致对特定基因网络异常如何导致人类神经精神疾病的深入了解; 这些见解对于理解病因、诊断和治疗至关重要。比如说， Tbr 1突变增加自闭症风险40. Tbr 1编码我们发现的TF， 2，3，13，38，41，42.我们假设，本文提出的工作将确定多个新的推定RE （pRE），包括Tbr 1，控制皮质发育，将有助于人类遗传学家定义 导致疾病风险的非编码突变的功能。

PROJECT SUMMARY (30 lines) Dysregulation of cortical development is central to epilepsy, mental deficiency, autism and schizophrenia. Cortical progenitors generate the projection neurons of the cerebral cortex and hippocampus. Understanding the genetic circuitry controlling the development and function of the cortex is essential to interpret human allele variants in people who have neuropsychiatric disorders. To elucidate the genetic circuitry driving the development of cortical progenitors and neurons, we must define the essential transcription factors (TF), regulatory elements (REs) and coding regions. The proposed research on cortical regionalization and laminar specification, will overcome these barriers through the identification and functional characterization of REs implicated in cortical development. Further, the regulation of REs by chromatin modifiers is critical to their appropriate spatiotemporal activity8,9. Here we propose to make inroads into these subjects. We aim to elucidate transcriptional mechanisms through which patterning of cortical progenitors is transmitted to, and maintained in, cortical neurons. In this proposal, we will study TFs and their targets (putative REs or pREs) regulating cortical patterning and lamination (Aim 1). We will profile the regional and laminar epigenomic states of pREs using purified cortical progenitors and differentiating neurons (Aims 2 & 3). We will define the activity and function of pREs using transgenic mice and enhancer deletions (Aim 4). Finally, to define the function of chromatin modifiers in cortical regionalization and lamination, we will study a chromatin modifier conditional mutant (Kdm6b cKO, Aim 5). The proposed studies shift our research from the function of single genes towards understanding how transcriptional networks orchestrate cortical development. We will integrate, our collaborative large-scale genomics analyses of forebrain pREs1,43,44, with our ChIP-Seq (Chromatin Immunoprecipitation-Sequencing) analyses, to identify regional and laminar specific cortical pREs. This information can lead to insights about human disease alleles in non-coding sequences. Moreover, our results will provide critical information about the genetic control of cortical progenitors and neurons. Once integrated with human genetic information, this will lead to insights into how abnormalities in specific gene networks cause human neuropsychiatric disorders; insights that are essential for understanding etiology, diagnosis and perhaps treatment. For instance, mutations in Tbr1 increase autism risk40. Tbr1 encodes a TF that we discovered and functionally characterized2,3,13,38,41,42. We hypothesize that the work proposed herein will identify multiple novel putative REs (pREs), including for Tbr1, that control cortical development, and will be helpful to human geneticists to define the function of non-coding mutations that contribute to disease risk.