Analysis of Glutamatergic Neurons Derived from Patient-Specific iPS Cells

患者特异性 iPS 细胞衍生的谷氨酸能神经元的分析

基本信息

批准号：
8121264
负责人：
HERBERT M LACHMAN
金额：
$ 41.5万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
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 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8121264
关键词：
22q11 Acute Erythroblastic Leukemia Address Agonist Automobile Driving Autopsy Biochemical Biocompatible Materials Bioinformatics Biological Assay Biological Psychiatry Bipolar Disorder Brain Budgets Cell Culture Techniques Cell Line Cells Chromosome abnormality Clinical Trials Commit Complementary DNA Complex Data Data Set Development Diagnosis Disease Electrophysiology (science)Enrollment Epigenetic Process Event Fibroblasts Functional Magnetic Resonance Imaging Functional disorder Funding Gender Gene Expression Gene Expression Profiling Genetic Heterogeneity Genome Glare Glutamates Goals Growth Guidelines Hair Root Heterogeneity Human Hybridization Array Image Impaired cognition In Vitro Individual Intervention Knockout Mice Lead Letters Leukocytes Libraries Life Mental disorders Methodology Methods Modeling Molecular Molecular Genetics Molecular Profiling Molecular and Cellular Biology Mus N-Methyl-D-Aspartate Receptors Neuroblastoma Neuroglia Neurons Oligodendroglia Pathogenesis Pathway interactions Patients Phase Predisposition Protocols documentation Publishing RNA Sequences Race Recruitment Activity Research Personnel Resistance Resources Sample Size Scheme Schizophrenia Serine Skin Source Specialist Specimen Stem cells Subgroup Supervision Symptoms Synapses System Time Tissues Work base brain tissue cell type clinically relevant cognitive function cohort disability embryonic stem cell experience genetic analysis genome wide association study genome-wide genome-wide analysis hippocampal pyramidal neuron induced pluripotent stem cell interest investigator training keratinocyte peripheral blood pluripotency public health relevance research study stem transmission process treatment trial

项目摘要

DESCRIPTION (provided by applicant): A number of studies suggest that schizophrenia (SZ) is caused by a deficiency in glutamatergic transmission. The analysis of glutamatergic pathways in SZ has been addressed using a variety of different experimental approaches, one of which is gene expression profiling using array hybridization carried out with postmortem brain tissue or peripheral blood leukocytes. These studies have provided many exciting clues to SZ pathogenesis. However, there are glaring limitations when leukocytes are used as a surrogate for gene expression in the brain, and many confounding factors make it difficult to interpret data from autopsy specimens. In addition, these sources of biological material cannot be used for electrophysiological analysis. Consequently, the ability to study molecular and electrophysiological events in cultivated patient-specific glutamatergic neurons would greatly enhance our understanding of underlying disease mechanisms. The discovery of induced pluripotent stem (iPS) cells, and the prospect of driving them to differentiate into glia, oligodendrocytes, and various neuronal subtypes, provides the opportunity to address the problems associated with interpreting data from postmortem tissue. We will generate iPS cells from hair root keratinocytes obtained from control subjects and patients with SZ. The patient cohort will include individuals with SZ who have 22q11 deletions, a genetically homogenous subgroup. Induced pluripotent stem cells will be driven to differentiate into glutamatergic pyramidal neurons using a protocol established in mouse ES cells, and expression profiling will be carried out using massively parallel deep sequencing (RNA-seq), which is more sensitive than array hybridization-based methods. Expression profiles between patients and controls will be compared to identify SZ-specific differences. Since the differentiation protocol being used generates glutamatertic pyramidal neurons capable of forming synaptic connections, electrophysiological analyses will be carried out as well. The proposed experiments using differentiating, patient-specific iPS cells will result in the development of a new method for studying the underlying molecular basis of schizophrenia. PUBLIC HEALTH RELEVANCE: This study describes our interest in growing neurons from patients with schizophrenia. The neurons will be generated from so-called induced pluripotent stem cells, which are derived from skin or hair roots. Induced pluripotent cells have the capacity to develop into many different cell types, including neurons. Studying neurons from patients with schizophrenia will contribute to our understanding of this enigmatic illness and possibly lead to new therapies.

描述（由申请人提供）：许多研究表明精神分裂症（SZ）是由谷氨酸能传递缺陷引起的。 SZ 中谷氨酸通路的分析已通过多种不同的实验方法进行了解决，其中之一是使用死后脑组织或外周血白细胞进行阵列杂交进行基因表达谱分析。这些研究为 SZ 发病机制提供了许多令人兴奋的线索。然而，当白细胞用作大脑中基因表达的替代物时，存在明显的局限性，并且许多混杂因素使得难以解释尸检标本的数据。此外，这些生物材料来源不能用于电生理学分析。因此，研究培养的患者特异性谷氨酸能神经元的分子和电生理学事件的能力将极大地增强我们对潜在疾病机制的理解。诱导多能干细胞 (iPS) 的发现，以及驱动它们分化为神经胶质细胞、少突胶质细胞和各种神经元亚型的前景，为解决与解释死后组织数据相关的问题提供了机会。我们将从对照受试者和 SZ 患者的发根角质形成细胞中产生 iPS 细胞。患者队列将包括 22q11 缺失的 SZ 个体，这是一个遗传同质的亚组。使用在小鼠 ES 细胞中建立的方案，诱导多能干细胞将被驱动分化为谷氨酸能锥体神经元，并且将使用大规模并行深度测序 (RNA-seq) 进行表达谱分析，这比基于阵列杂交的方法更灵敏。将比较患者和对照之间的表达谱，以确定 SZ 特异性差异。由于所使用的分化方案产生能够形成突触连接的谷氨酸锥体神经元，因此也将进行电生理学分析。拟议的使用分化的、患者特异性 iPS 细胞的实验将开发出一种新方法来研究精神分裂症的分子基础。公共健康相关性：这项研究描述了我们对精神分裂症患者神经元生长的兴趣。这些神经元将由所谓的诱导多能干细胞产生，这些干细胞源自皮肤或发根。诱导多能细胞有能力发育成许多不同的细胞类型，包括神经元。研究精神分裂症患者的神经元将有助于我们了解这种神秘的疾病，并可能带来新的治疗方法。