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

• 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
• 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

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.

总结 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。 紊乱我们已经发现了一种新的压力调节途径， 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大脑中的失调，以及治疗双相情感障碍患者的方法 与锂，一种激活休息的药物，恢复体内平衡。该项目将解决一个新的概念 这两种疾病的基础可能都涉及大脑中应激反应系统的失调 这是由REST管理的。最初，我们将确定是否REST功能在来自于神经元的神经元中发生改变。 患者的诱导祖干（iPS）细胞和我们引入了诱导祖干细胞的同基因iPS细胞系 使用CRISPR-Cas9系统，研究导致精神分裂症的遗传变体。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 在单神经元分辨率下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。完成 为了实现这一目标，我们将推进一项我们最近开发的新技术，称为原位荧光技术。 RNA测序（FISSEQ）。与需要分离DNA或DNA片段的常规测序相比， RNA，FISSEQ序列完整组织中的RNA，汇集了转录组范围内RNA的深度 具有单分子原位RNA定位分辨率的测序。FISSEQ也可以多路复用 与其他数据流，特别是蛋白质组学，使单细胞多维询问， 分辨率FISSEQ将用于推导人类中已鉴定神经细胞类型的参考转录组， 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，并提供了一个系统- 中央调控通路的水平视图。这将通过小鼠中的转录组分析来补充 基因工程化以在特定神经元群体中缺失或过表达REST基因。虽然这 这种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的 研究人员与互补领域的专业知识，在一个多学科的方法，以了解精神病 疾病和先进的大脑单细胞转录组分析。

项目成果

FACS-Based Sequencing Approach to Evaluate Cell Type to Genotype Associations Using Cerebral Organoids.

基于 FACS 的测序方法使用脑类器官评估细胞类型与基因型关联。

• DOI: 10.1007/978-1-0716-3287-1_15
• 发表时间: 2023
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Murray,Liam;Olson,MeaganN;Barton,Nathaniel;Dawes,Pepper;Chan,Yingleong;Lim,ElaineT
• 通讯作者: Lim,ElaineT