Describing the Networks of Disease-associated Transcription Factors in the Brain.

项目摘要 精神分裂症是一种严重的精神疾病，影响世界人口的百分之一。虽然 已经表明，这种疾病可能与神经发育过程中的分子变化有关， 这些变化的驱动因素仍然未知。转录因子（TF）是进一步研究转录因子的有吸引力的候选者。 研究;他们控制整个基因网络的表达，广泛的TF驱动的网络控制着基因的表达。 神经元和其他脑细胞类型的发育。我们之前的工作利用了一个公开可用的GWAS， 精神分裂症和大脑中可接近的染色质图谱，以建立一个调节组范围的关联模型， 这种疾病的。该模型使用DNAse-seq“足迹”来确定TF的可能结合位点， 来预测靶基因。具有许多与GWAS风险相关的靶点的TF被认为是 与自己的风险有关。这使我们能够鉴定出可能参与糖尿病发病机制的转录因子。 精神分裂症，以及这些TF的个体靶基因。在我们的模型中识别的许多顶级TF 已知参与细胞命运决定，包括称为EMX 1的神经发育TF。我们有 独立观察到精神分裂症病例与对照组相比EMX 1结合活性增加，并显示 EMX 1靶向表达在兴奋性神经元谱系中增加。这项工作旨在验证我们的模型 并测试了精神分裂症与转录网络变化相关的假设， EMX 1推动神经前体过早分化为兴奋性神经元。计算机模拟- 使用高质量ATAC-seq和单细胞RNA-seq数据集（目标1）的方法将用于确认我们的 先前的研究结果表明，EMX 1显示出增加的结合活性，并表明其结合靶点参与了 兴奋性神经元的发育胚胎干细胞衍生的神经诱导系统（Aim 2）将在 用于验证我们的模型预测的EMX 1靶点，并证明EMX 1过表达是如何相互作用的。 发热伴皮层形成。这一目标将检验EMX 1的过度表达激活了一个净- 在皮质生成过程中，靶基因的工作导致兴奋性神经元过早形成，导致后来的 皮质结构和厚度的缺陷。这些实验将证明我们的模型在识别中的实用性。 TF网络介导了精神分裂症的风险，并证明了EMX 1网络在神经系统中的作用。 细胞类型确定。这个项目是一个更大的培训计划的一部分，旨在培养我的技能和 作为一名神经精神系统生物学领域的内科医生和科学家所必需的技术。我会执行 在我的主要赞助人Seth Ament博士和我的共同赞助人Margaret博士的指导下， 麦卡锡Ament博士是神经精神疾病系统生物学方面的专家， 超过27篇论文。McCarthy博士是神经发育领域的杰出研究人员， 博士前学生的培训经验，包括MD/博士生。这个指导小组将监督我的 随着我实现这份提案的目标和我更广泛的职业目标，我将努力发展成为一名科学家和临床医生。

PROJECT SUMMARY Schizophrenia is a serious psychiatric disorder that affects up to one percent of the world's population. While it has been shown that this disease may be associated with molecular changes during neurodevelopment, the drivers of these changes remains unknown. Transcription factors (TFs) are attractive candidates for further study; they control the expression of entire networks of genes, and extensive TF-driven networks govern the development of neurons and other brain cell types. Our previous work leverages a publicly available GWAS of schizophrenia and an atlas of accessible chromatin in the brain to establish a regulome-wide association mod- el of this disease. This model uses DNAse-seq “footprints” to determine likely binding sites of TFs and uses these sites to predict target genes. TFs with many targets associated with risk in GWAS are considered asso- ciated with risk themselves. This has allowed us to identify TFs that may be involved in the pathogenesis of schizophrenia, as well as the individual target genes of these TFs. Many of the top TFs identified in our model are known to be involved in cell-fate decisions, including a neurodevelopmental TF known as EMX1. We have independently observed increases in EMX1 binding activity in schizophrenia cases versus controls, and shown that EMX1 target expression is increased in excitatory neuron lineages. This work aims to validate our model and test the hypothesis that schizophrenia is associated with changes in a transcriptional network governed by EMX1 that pushes neural precursors towards premature differentiation into excitatory neurons. In silico ap- proaches using high-quality ATAC-seq and single cell RNA-seq datasets (Aim 1) will be used to confirm our previous findings that EMX1 shows increased binding activity and show that its binding targets are involved in the development of excitatory neurons. An embryonic stem cell-derived neural induction system (Aim 2) will be used to validate targets of EMX1 predicted by our model and demonstrate how EMX1 overexpression inter- feres with cortex formation. This aim will test the hypothesis that the overexpression of EMX1 activates a net- work of target genes that causes premature excitatory neuron formation during corticogenesis, resulting in later deficits in cortical structure and thickness. These experiments will demonstrate the utility of our model in identi- fying TF networks that mediate risk for schizophrenia, and demonstrate the role of the EMX1 network in neural cell type determination. This project is a part of a larger training plan designed to train me in the skills and techniques necessary for a career as a physician-scientist in neuropsychiatric systems biology. I will carry out this proposal under the guidance of my primary sponsor, Dr. Seth Ament, and my co-sponsor Dr. Margaret McCarthy. Dr. Ament is an expert in the systems biology of neuropsychiatric disease and has published more than 27 papers on the subject. Dr. McCarthy is a pre-eminent researcher in neurodevelopment with extensive experience training pre-doctoral students, including MD/PhD students. This mentoring team will supervise my development as a scientist and a clinician as I achieve the aims of this proposal and my broader career goals.