Integrated, cell type specific functional genomics analyses of regulatory sequence elements and their dynamic interaction networks in neuropsychiatric brain tissues

项目概要/摘要 经过一个世纪关于神经精神疾病基本性质的争论，我们知道遗传学会撒谎 他们的核心，但不完全了解其致病病理学的关键基本机制。 我们建议的总体目标是建立全面和综合的染色质图谱 可及性、染色体折叠和转录模式，描绘基因组中的调控区域 成人和胎儿大脑的关键疾病相关解剖区域，精神分裂症患者的大脑， 自闭症谱系障碍，双相情感障碍和匹配的对照，以及那些已知的CNVs （拷贝数变异），可能揭示表观基因组调控相互作用的区域或远程靶标 这在具有相同临床表型的患者中也可能是非常相关的。我们将全面 以及我们最近开发的高分辨率表观基因组学分析，以及整合这些基因的新方法。 这是第一次在神经精神病学相关的脑组织中获得数据。我们将生成全面的地图 通过整合互补技术，调整区域的组织和功能范围： 单细胞ATAC-seq（scATAC-seq）表征染色质开放性，HiChIP表征长 分选的神经元和非神经元细胞的范围折叠相互作用，两者都偶联到单细胞 RNA-seq和长距离RNA-seq用于表达信息，进一步补充了关于 转录因子通过蛋白质组学分析核组分。这些地图将与 在候选区域中的基因组序列中的编码或非编码/调节变体，并整合到 整个PsychENCODE数据库，这将使我们能够创建和验证表观基因组的参考图谱， 标记和相互作用，确定患者组织中与参考状态的偏差，并将这些 遗传疾病基因座的畸变以及将这些基因座组装成疾病途径。这项工程计划不会 这只会大大扩展我们对人类基因组中编码的调控信息及其影响的理解， 研究人类大脑发育和神经精神疾病，还产生了生物信息学工具， 分析PsychENCODE中生成的复杂数据所必需的。

Project Summary/Abstract After a century of debate about the fundamental nature of neuropsychiatric disorders, we know that genetics lie at their core, yet do not fully understand the critical underlying mechanisms of their disease-causing pathology. The overall goal of our proposal is the creation of comprehensive and integrated maps of chromatin accessibility, chromosome folding and transcriptional patterns, delineating regulatory regions in the genomes of key disease relevant anatomical regions of adult and fetal brains, in brains from patients with Schizophrenia, Autism Spectrum Disorder, Bipolar Affective Disorder and matched controls, and those with known CNVs (Copy-Number Variants) that may unmask regional or long-range targets of epigenomic regulatory interactions that may also be of great relevance in patients with the same clinical phenotype. We will use comprehensive and highly-resolving epigenomics assays, that were recently developed by us, and novel ways to integrate the data for the first time in neuropsychiatrically relevant brain tissues. We will generate comprehensive maps of the spectrum of organization and function of regulatory regions by integrating complementary techniques: single-cell ATAC-seq (scATAC-seq) to characterize chromatin openness and HiChIP to characterize long- range folding interactions of sorted neuronal and non-neuronal cells, both of which are coupled to single-cell RNA-seq and long-range RNA-seq for expression information, further complimented by information about transcription factors through proteomic analysis of nuclear fractions. These maps will then be combined with coding or non-coding/regulatory variants in the genomic sequence in the candidate regions and integrated into the overall PsychENCODE database, which will allow us to create and validate reference maps for epigenomic marks and interactions, determine aberrations to the reference state in patient tissue, and connect such aberrations to genetic disease loci as well as assemble such loci into disease pathways. This project will not only greatly expand our understanding of regulatory information encoded in the human genome and its impact on human brain development and neuropsychiatric disorders, but also produce the bioinformatics tools necessary to analyze the complex data being generated in PsychENCODE.