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: 9260728
• 负责人: DOUGLAS Frederick LEVINSON
• 金额: $ 29.64万
• 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/9260728
• 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; Computer Analysis; DNA Sequence Alteration; Defect; Derivation procedure; Development; Disease; Electrophysiology (science); Engineering; Equilibrium; Functional disorder; Future; Generations; Genes; Genetic; Genomics; Genotype; Goals; Health; 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; conditional mutant; 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; 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 项目的目标是“与学术界和工业界合作，创建多学科研究小组，利用患者来源的重编程细胞开发经过验证的平台，以识别新靶标并开发新疗法……以减轻精神疾病的负担。”在这里，我们建议使用源自诱导多能干 (iPS) 细胞的诱导神经元 (iN) 细胞来模拟突触功能缺陷，这些缺陷是由于已知会显着增加精神分裂症 (SCZ) 风险的三种工程或自然发生的突变所致：NRXN1 外显子缺失、22q11.2 缺失和 16p11.2 重复。该研究包括三个项目。项目1将分析人类iN细胞的功能特征。对于每种突变，一个组件将研究来自同一对照个体的带有工程突变的细胞与非突变细胞，而第二个组件将研究源自 5 名具有感兴趣突变的 SCZ 患者与 5 名对照个体的 iN 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，并尝试确定这些突变的突触表型在 iN 细胞、培养的原代神经元和脑切片（医学前额叶皮层）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iN细胞方案；针对大型CNV的新的靶向突变策略；纯抑制性iN细胞的衍生；开发无小鼠的iN细胞方案），并且还需要根据本研究中开发的病理生理学模型开发未来的高通量筛选测定法。这些实验将为通过重编程方法衍生的神经元特征、每种突变产生的突触表型以及最终对包括 SCZ 在内的精神病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或至少部分重叠的突触表型。这两种观察结果都对未来 SCZ 研究具有深远的影响。为了完成这项工作，我们组建了一支来自斯坦福大学的优秀科学团队（分子神经科学领域的 Sudhof 博士、干细胞生物学领域的 Wernig 博士和精神分裂症遗传学领域的 Levinson 博士）；罗格斯大学（庞博士，神经科学和干细胞生物学）；辛辛那提儿童医院医疗中心（Aronow 博士，生物信息学和高内涵成像）；礼来公司（Isaac 和 Ursu 博士负责电生理学，Merchant 博士负责药物开发，Dage 博士负责高内涵成像和分析开发，Collier 博士负责基因组学和系统生物学，Eastwood 博士负责生物统计学）； Cellular Dynamics, Inc.（Swanson 博士，代表干细胞生物学领域领先的生物技术公司）。这些项目代表了学术和工业机构的多学科努力，旨在通过研究与 SCZ 相关的三种突变来深入了解精神障碍的病理生理学。