Multimodal analysis of high-risk psychosis mutations in induced neuronal cells

诱导神经元细胞高危精神病突变的多模态分析

• 批准号: 8743628
• 负责人: DOUGLAS Frederick LEVINSON
• 金额: $ 209.34万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-09 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8743628
• 关键词: 16p11.2; 22q11.2; Academia; Bioinformatics; Biological Assay; Biometry; Biotechnology; Blood; Brain; Cell Maturation; Cell surface; Cells; Characteristics; Chemicals; Clinical; Coculture Techniques; Complex; Defect; Derivation procedure; Development; Disease; Electrophysiology (science); Engineering; Equilibrium; Functional disorder; Future; Gene Mutation; Generations; Genes; Genetic; Genomics; Genotype; Goals; Human; Human Characteristics; Image; Individual; Industry; Institution; Interdisciplinary Study; Measures; Mediating; Medical; Medical center; Mental disorders; Methodology; Methods; Modality; Modeling; Molecular; Mus; Mutation; National Institute of Mental Health; Neuroglia; Neurons; Neurosciences; Odds Ratio; Patients; Pediatric Hospitals; Phenotype; Predisposition; Primary Cell Cultures; Procedures; Production; Protocols documentation; Psychotic Disorders; Reproducibility; Research; Risk; Sampling; Scanning; Schizophrenia; Slice; Stem cells; Synapses; Systems Biology; T-Lymphocyte; Testing; Therapeutic; Time; Universities; Validation; Work; assay development; base; brain cell; design; drug development; frontal lobe; high risk; high throughput screening; induced pluripotent stem cell; insight; interest; mouse model; multidisciplinary; novel; novel therapeutics; portability; presynaptic; programs; public health relevance; repository; research study; signal processing; stem cell biology; synaptic function; transmission process

DESCRIPTION (provided by applicant): The goal of the NCRCRG program is "to create multidisciplinary research groups, in partnership with academia and industry, to use patient-derived reprogrammed cells to develop validated platforms for identifying novel targets and developing new therapeutics ... to reduce the burden of mental illness." Here we propose to use induced neuronal (iN) cells, derived from induced pluripotent stem (iPS) cells, to model defects in synaptic function due to three engineered or naturally-occurring mutations known to substantially increase the risk of schizophrenia (SCZ): NRXN1 exonic deletions, 22q11.2 deletions, and 16p11.2 duplications. The study includes three projects. Project 1 will analyze the functional characteristics of human iN cells. For each mutation, one component will study cells with engineered mutations vs. non-mutated cells from the same control individual, while a second component will study iN cells derived from 5 SCZ patients with the mutation of interest vs. 5 control individuals. Project 2 will analyze the same mutations in mouse models, providing both a cross-species validation of human findings, and an novel attempt to determine whether the synaptic phenotypes of these mutations are the same in iN cells, cultured primary neurons, and brain slices (medical pre-frontal cortex). Project 3 will develop and optimize stem cell methods that are required for this project (large-scale implementation of iN cell protocols; new targeted mutation strategies for large CNVs; derivation of pure inhibitory iN cells; development of a mouse-free iN cell protocol), and which will also be needed to develop future high-throughput screening assays based on the pathophysiological models developed in this study. These experiments will provide new insights into the characteristics of neurons derived by reprogramming method, into synaptic phenotypes produced by each of these mutations, and ultimately into the pathophysiology susceptibility to the risk of psychotic disorders including SCZ. We will determine whether these mutations produce distinct or at least partially overlapping synaptic phenotypes. Either observation has profound implications for future SCZ research. To accomplish this work, we have assembled an outstanding scientific team from Stanford University (Drs. Sudhof in molecular neuroscience, Wernig in stem cell biology and Levinson in genetics of schizophrenia); Rutgers University (Dr. Pang in neuroscience and stem cell biology); Cincinnati Children's Hospital Medical Center (Dr. Aronow in bioinformatics and high-content imaging); Eli Lilly and Company (Drs. Isaac and Ursu in electrophysiology, Merchant in drug development, Dage in high-content imaging and assay development, Collier in genomics and systems biology, and Eastwood in biostatistics); and Cellular Dynamics, Inc. (Dr. Swanson, representing a leading biotechnology company in stem cell biology). These projects represent a multidisciplinary effort of academic and industrial institutions to gain insight into te pathophysiology of psychotic disorders by studying three mutations that are associated with SCZ.

描述(由申请人提供)：NCRCRG计划的目标是“与学术界和工业界合作，创建多学科研究小组，利用患者来源的重新编程细胞开发有效的平台，以确定新的靶点和开发新的治疗方法……以减轻精神疾病的负担。”在这里，我们建议使用来自诱导多能干细胞(IPS)的诱导神经元(IN)细胞来模拟突触功能缺陷，这些缺陷是由于三种已知会显著增加精神分裂症(SCZ)风险的工程或自然发生的突变：NRXN1外显子缺失、22q11.2缺失和16p11.2重复。这项研究包括三个项目。项目1将分析人类在细胞中的功能特征。对于每个突变，一个组件将研究具有工程突变的细胞与来自同一对照个体的非突变细胞，而第二个组件将研究来自5名感兴趣突变的SCZ患者与5名对照个体的细胞。项目2将分析小鼠模型中的相同突变，提供对人类发现的跨物种验证，并提供一种新的尝试，以确定这些突变的突触表型在细胞、培养的初级神经元和脑片(医学前额叶皮质)中是否相同。项目3将开发和优化该项目所需的干细胞方法(大规模实施细胞内协议；针对大型CNV的新的靶向突变策略；获得细胞内的纯抑制作用；开发无鼠的细胞内协议)，并且还将需要这些方法来基于本研究中开发的病理生理模型开发未来的高通量筛选分析。这些实验将为通过重新编程方法获得的神经元的特征、由每个突变产生的突触表型以及最终对包括SCZ在内的精神障碍风险的病理生理学易感性提供新的见解。我们将确定这些突变是否产生不同的或至少部分重叠的突触表型。这两个观察结果都对未来的SCZ研究有深远的影响。为了完成这项工作，我们汇聚了一支杰出的科学团队，他们来自斯坦福大学(分子神经科学方面的苏德夫博士、干细胞生物学方面的Wernig博士和精神分裂症遗传学方面的Levinson博士)、罗格斯大学(神经科学和干细胞生物学方面的彭博士)、辛辛那提儿童医院医学中心(生物信息学和高内容成像方面的Aronow博士)、礼来公司(电生理学方面的Isaac和Ursu博士、药物开发方面的Merchant博士、高含量成像和化验开发方面的Dage博士、基因组学和系统生物学方面的Collier博士以及生物统计学方面的Eastwood博士)；和Ccell Dynamic，Inc.(代表干细胞生物学领域领先生物技术公司的Swanson博士)。这些项目代表了学术和工业机构的多学科努力，通过研究与SCZ相关的三个突变来深入了解精神障碍的病理生理学。