Exploring a Novel Paradigm of Schizophrenia and Bipolar Disorder

探索精神分裂症和双相情感障碍的新范式

基本信息

批准号：
9981018
负责人：
GEORGE M CHURCH
金额：
$ 93.63万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-26 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9981018
关键词：
Address Antipsychotic Agents Area Behavioral Biological Biological Models Bipolar Disorder Brain Brain Diseases CRISPR/Cas technology Cell Line Cells Classification Clinical Clinical Treatment Complement Complex DNA Delusions Development Diagnostic Disease Disease Pathway Fluorescence Functional disorder Genes Genetic Transcription Genetically Engineered Mouse Genome Glutamates Goals Hallucinations Homeostasis Human In Situ Knockout Mice Lead Lithium Mediating Mental disorders Metabolic Modernization Mus Neurobehavioral Manifestations Neurology Neurons Neurosciences Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Physiological Play Population Principal Investigator Proteomics Psychiatry RNA RNA Sequences Regulation Regulatory Pathway Resolution Role Schizophrenia Stress Symptoms System Technology Therapeutic Tissues Transcription Repressor Transgenic Mice base biological adaptation to stress bipolar patients brain cell cell type conditional knockout data streams druggable target genetic risk factor genetic variant in situ sequencing innovation insight interdisciplinary approach mouse model neural circuit neurotransmission new technology novel overexpression progenitor relating to nervous system single molecule stem cells symptomatology therapeutic target transcription factor transcriptome transcriptome sequencing

项目摘要

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.

概括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疾病之一失调。我们发现了一种由大师介导的大脑压力调节的新途径转录抑制因子 REST/NRSF。我们的初步研究表明，REST 途径可能是精神分裂症和双相情感障碍患者的大脑失调以及治疗双相情感障碍患者锂是一种激活休息、恢复体内平衡的药物。该项目将提出一个新颖的概念可能是涉及大脑应激反应系统失调的两种疾病的基础的范例由 REST 监管。首先，我们将确定源自神经元的 REST 功能是否发生改变患者的诱导祖干细胞和同基因 iPS 细胞系中，我们在其中引入了使用 CRISPR-Cas9 系统研究导致精神分裂症的遗传变异。然后我们将确定是否有条件特定神经元亚群中 REST 功能障碍的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，我们将以单神经元分辨率定义精神分裂症和双相情感障碍患者大脑中的转录组变化确定 REST 和其他转录调节因子在特定神经元细胞类型中的作用。为了完成为了这个目标，我们将推进我们最近开发的一项新技术，称为原位荧光 RNA 测序 (FISSEQ)。与传统测序不同的是，传统测序需要分离 DNA 或 RNA、FISSEQ 对完整组织中的 RNA 进行测序，汇集了全转录组 RNA 的深度具有单分子原位RNA定位分辨率的测序。 FISSEQ 也可以进行复用与其他数据流，特别是蛋白质组学，实现单细胞的多维询问解决。 FISSEQ 将用于衍生已识别的人类神经细胞类型的参考转录组和老鼠的大脑。参考转录组和患者转录组之间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，并提供系统- 中央监管途径的水平视图。这将通过小鼠转录组分析得到补充通过基因工程删除或过度表达特定神经元群体中的 REST 基因。虽然这该方法将用于探索精神疾病，一旦开发出来，它可以迅速用于阐明任何脑部疾病中基因组调控的改变。这些研究将汇集两个主要具有互补专业知识领域的研究人员，采用多学科方法来了解精神病学疾病和先进的大脑单细胞转录组分析。