Exploring a Novel Paradigm of Schizophrenia and Bipolar Disorder

摘要 精神分裂症和双相情感障碍是临床上定义最多但了解最少的精神病学 精神错乱。我们已经在大脑中发现了一种新的压力调节途径，这种调节是由大师调节的。 转录抑制因子REST/NRSF。我们的初步研究表明，休息途径可能是 精神分裂症和双相情感障碍患者的大脑调节失调以及对双相情感障碍患者的治疗 锂是一种激活休息的药物，可以恢复体内平衡。这个项目将解决一个新的概念 可能导致这两种疾病的范例，涉及大脑中应激反应系统的失调 这是由休息来调节的。最初，我们将确定是否改变了来自 患者的诱导性祖细胞(IPS)以及我们已经在其中引入了 使用CRISPR-CAS9系统的导致精神分裂症的基因变异。然后我们将确定是否有条件 特定神经元亚群中静息功能障碍的基因敲除小鼠模型概括了行为， 与精神分裂症和双相情感障碍相关的代谢和生理变化。最后，我们会 定义精神分裂症和双相情感障碍患者大脑在单个神经元分辨率下的转录组变化 确定REST和其他转录调节因子在特定神经细胞类型中的作用。要完成 为了实现这一目标，我们将推进我们最近开发的一项新技术--原位荧光 RNA测序(FISSEQ)。与传统测序相反，传统测序需要分离DNA或 RNA，FISSEQ序列完整组织中的RNA，汇集了整个转录组的深度RNA 单分子原位RNA定位的分辨率测序。FISSEQ也可以被多路传输 利用其他数据流，特别是蛋白质组学，实现了单细胞的多维询问 决议。FISSEQ将用于获得人类已识别神经细胞类型的参考转录本 和老鼠的大脑。慢性粒细胞白血病患者参考转录本与转录本的差异 精神分裂症和双相情感障碍可能涉及REST或其他转录因子，并提供一种系统-- 中央调控途径的水平视图。这将得到小鼠转录组分析的补充。 通过基因工程在特定的神经元群体中删除或过度表达REST基因。虽然这件事 这种方法将被用来探索精神障碍，一旦开发出来，它可以迅速应用于 阐明在任何大脑疾病中改变的基因组调控。这些研究将汇集两个主要因素 在多学科方法方面具有互补专业知识以了解精神病学的调查人员 疾病和先进的大脑单细胞转录组分析。

Summary Schizophrenia and bipolar disorder are among the most clinically defined but least understood psychiatric disorders. We have uncovered a novel pathway of stress regulation in the brain mediated by the master transcriptional repressor REST/NRSF. Our preliminary studies suggest that the REST pathway may be dysregulated in the brains of patients with schizophrenia and bipolar disorder, and that treating bipolar patients with lithium, a drug that activates REST, restores homeostasis. This project will address a novel conceptual paradigm that might underlie both disorders involving dysregulation of a stress response system in the brain that is regulated by REST. Initially, we will determine if REST function is altered in neurons derived from induced progenitor stem (iPS) cells of patients and in isogenic iPS cell lines in which we have introduced a schizophrenia-causing genetic variant using the CRISPR-Cas9 system. We will then determine if conditional knockout mouse models of REST dysfunction in specific neuronal subpopulations recapitulate behavioral, metabolic and physiological changes associated with schizophrenia and bipolar disorder. Finally, we will define transcriptome changes in the brains of schizophrenic and bipolar patients at single neuron resolution to ascertain the role of REST and other transcriptional regulators in specific neuronal cell types. To accomplish this goal, we will advance a new technology we have recently developed called fluorescence in situ sequencing of RNA (FISSEQ). In contrast to conventional sequencing, which requires isolation of DNA or RNA, FISSEQ sequences RNA in intact tissue, bringing together the depth of transcriptome-wide RNA sequencing with the resolution of single molecule in situ RNA localization. FISSEQ can also be multiplexed with other data streams, particularly proteomics, enabling multidimensional interrogation at single cell resolution. FISSEQ will be used to derive reference transcriptomes for identified neural cell types in the human and mouse brain. Differences between reference transcriptomes and transcriptomes of patients with schizophrenia and bipolar disorder may implicate REST or other transcription factors, and provide a systems- level view of the central regulatory pathways. This will be complemented by transcriptome analysis in mice genetically engineered to delete or overexpress the REST gene in specific neuronal populations. Although this approach will be used to explore psychiatric disorders, once developed, it could be rapidly employed to elucidate altered genome regulation in any brain disorder. These studies will bring together two principal investigators with complementary areas of expertise in a multidisciplinary approach to understand psychiatric disorders and advance single cell transcriptome analysis of the brain.