Genomics-Guided Characterization of iPS Cells from Common Mental Illnesses

常见精神疾病 iPS 细胞的基因组学引导表征

基本信息

批准号：
8206071
负责人：
STEPHEN J HAGGARTY
金额：
$ 44.08万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8206071
关键词：
22q11 Accounting Address Adult Affect Alleles Ankyrins Antibodies Automobile Driving Axon Biological Assay Biopsy Bipolar Disorder Brain Diseases Brain-Derived Neurotrophic Factor Cell Line Cell model Cells Child Chromosomal Stability Chromosome Deletion Chromosomes Clinical Collection Complex Consent Core Facility DNA DNA Fingerprinting Databases Development Diagnosis Disease Disease Progression Disease susceptibility Documentation Ensure Equipment and supply inventories Etiology Family Fibroblasts Fingerprint Functional disorder Gene Delivery General Hospitals Generations Genes Genetic Genetic Epistasis Genetic Risk Genetic Variation Genome Genomics Genotype Glycogen Synthase Kinase 3 Goals Haplotypes Heritability Human Human Genetics Image In Vitro Individual Institutes Institutional Review Boards Ion Channel Karyotype determination procedure Libraries Lithium Massachusetts Measurement Mediating Mental Depression Mental disorders Mentally Ill Persons Methodology Methods Microscopy Modality Modeling Molecular NRG1 gene Neuroglia Neurons Nuclear Family Nucleotides Parents Pathway interactions Patients Phase Phenotype Physiology Procedures Property Proteins Protocols documentation RNA Reader Reporter Genes Reporting Research Resources Reverse Transcriptase Polymerase Chain Reaction Risk Sampling Schizophrenia Siblings Signal Pathway Signal Transduction Single Nucleotide Polymorphism Skin Societies Somatic Cell Stem Cell Research Stem cells Study models Testing Tissues Twin Multiple Birth Variant Viral Work base c-myc Genes cell type disorder risk embryonic stem cell experience fluorescence imaging genetic pedigree genome wide association study high risk imaging modality induced pluripotent stem cell inhibitor/antagonist lymphoblast member miniaturize neural circuit neurodevelopment neuron development neuropsychiatry neurotrophic factor new therapeutic target novel novel strategies pluripotency public health relevance relating to nervous system response self-renewal small molecule software systems stem stem cell technology structural genomics success

项目摘要

DESCRIPTION (provided by applicant): Bipolar disorder and schizophrenia are significant burdens to patients, families and society. Despite consistent evidence for high heritability (up to 80%), the etiology remains poorly understood. In 2008, several groups including ours observed that copy number variants, and in particular, large deletions each containing many genes accounted for some fraction of disease susceptibility. Several groups also identified a more general excess of rare CNVs in schizophrenia and more recently, common single nucleotide variation has been shown to contribute to a polygenic component that also accounts for a fraction of disease risk. Finally, several statistically compelling specific risk loci have also been reported. With what is proving to be a highly complex genetic landscape, existing approaches that model changes in single or small numbers of genes across development are not well suited to capture all the genetic factors interacting in an individual. Thus, the overall goal of this project is to develop human induced pluripotent stem (iPS) cell-based models of neuropsychiatric disorders that are genetically based. In the R21 Phase, pre-existing publicly available fibroblasts from bipolar disorder and schizophrenia patients along with healthy controls will be used to pilot human iPS cell model methods. Once generated, methods for the differentiation of these iPS cells into neural lineages will be employed to characterize potential between-subject and within-subject differences. Specific signaling pathways previously implicated in psychiatric disease will be characterized using a panel of neurotrophic factors and small-molecule probes of Wnt/GSK-3 signaling to develop automated microscopy- based imaging and pathway-selective reporter gene assays. Upon completion of these studies we will have established a framework for systematic development of a Mental Illness Stem Cell Library (MISCL) that will be expanded in the second R33 Phase to include samples obtained from patients from our ongoing genome-wide association studies (GWAS) of bipolar disorder and schizophrenia. The driving questions in the R33 phase will be to use iPS cells derived from clinical samples that reflect the entire genome of an individual to determine the effects on neurodevelopment of: 1) large multigenic deletions on chromosome 1q21.1, 22q11 or 15q13.1 and 2) multiple polygenes of small effect. Similarly, we will use iPS cells from individuals with bipolar disorder associated ankyrin-G (ANK3) haplotypes to determine the effect on basic ion channel physiology, molecular organization of the axon initial segment, and neuronal polarity. PUBLIC HEALTH RELEVANCE: The purpose of this Project is to use newly developed methods for reprogramming human somatic cells (e.g., fibroblasts) to create induced pluripotent stem (iPS) cells that can be used as genetically accurate models of bipolar disorder and schizophrenia. To do so we will work closely with the Harvard Stem Cell Institute (HSCI) iPS Core facility at Massachusetts General Hospital, which has extensive experience with human iPS cell technologies and have successfully derived iPS lines from adult fibroblasts. Methods for inducing the differentiation of iPS cells into neural and glial cells will be implemented as well as novel methods for phenotyping cells using imaging and plate reader assays.

描述（由申请人提供）：双相情感障碍和精神分裂症对患者，家庭和社会来说是巨大的负担。尽管始终如一地证明了高遗传力（多达80％），但病因仍然知之甚少。在2008年，包括我们的几个小组观察到拷贝数变异，尤其是大的缺失，每个缺失都包含许多基因，这些基因占疾病易感性的一定程度。几个小组还确定了精神分裂症中罕见的CNV的过量过量，最近，常见的单核苷酸变异已被证明有助于多基因成分，该成分也占疾病风险的一部分。最后，还报道了一些统计上令人信服的特定风险基因座。由于事实证明是一种高度复杂的遗传景观，现有的方法在整个发育中的单个或少数基因中变化的现有方法并不适合捕获个体中的所有遗传因素。因此，该项目的总体目标是开发基于基于遗传的神经精神疾病的基于人类诱导的多能茎（IPS）细胞模型。在R21阶段，来自双相情感障碍和精神分裂症患者以及健康对照组的预先存在的公开可用的成纤维细胞将用于驾驶人IPS细胞模型方法。一旦生成，将采用将这些IPS细胞分化为神经谱系的方法来表征受试者间和受试者内部差异。先前与精神病有关的特定信号通路将使用一组神经营养因子和Wnt/GSK-3信号的小分子探针来表征，以开发基于显微镜的成像和途径选择性潜伏基因基因。这些研究完成后，我们将建立一个用于系统发展的精神疾病干细胞文库（MISCL）的框架，该框架将在第二R33阶段扩展，其中包括从我们正在进行的全基因组关联研究（GWAS）的患者中获得的样本（GWAS）和精神分裂症。 R33阶段的驱动问题将是使用源自临床样本的IPS细胞，这些临床样品反映了一个个体的整个基因组来确定以下方面的神经发育的影响：1）大型多基因缺失对1q21.1、22q11或15q113.1和15q13.1和2）的大型多基因缺失。同样，我们将使用来自与躁郁症的个体相关的Ankyrin-G（ANK3）单倍型的IPS细胞来确定对碱性离子通道生理学的影响，轴突初始段的分子组织和神经元极性。公共卫生相关性：该项目的目的是使用新开发的方法来重编程人类体细胞（例如成纤维细胞）来创建诱导的多能茎（IPS）细胞，这些细胞（IPS）可以用作双相情感障碍和精神分裂症的遗传准确模型。为此，我们将与马萨诸塞州综合医院的哈佛干细胞研究所（HSCI）IPS核心设施紧密合作，该医院在人类IPS细胞技术方面拥有丰富的经验，并成功地从成人成纤维细胞中得出了IPS系列。将实施用于诱导IPS细胞分化为神经细胞和神经胶质细胞的方法，以及使用成像和读取器分析的新方法来进行表型细胞。