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

• 批准号: 8124996
• 负责人: HERBERT M LACHMAN
• 金额: $ 39.89万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8124996
• 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; Cellular biology; 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 Biology; Molecular Genetics; Molecular Profiling; Mus; N-Methyl-D-Aspartate Receptors; Neuroblastoma; Neuroglia; Neurons; Oligodendroglia; Pathogenesis; Pathway interactions; Patients; Phase; Predisposition; Protocols documentation; Publishing; RNA; 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; 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

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）是由谷氨酸能传播缺乏引起的。已经使用多种不同的实验方法来解决SZ中谷氨酸能途径的分析，其中一种是使用使用后死后脑组织或外周血白细胞进行的阵列杂交进行基因表达分析。这些研究为SZ发病机理提供了许多令人兴奋的线索。但是，当用白细胞用作大脑基因表达的替代物时，存在明显的局限性，许多混杂因素使得很难解释尸检标本中的数据。此外，这些生物材料的来源不能用于电生理分析。因此，在培养的患者特异性谷氨酸能神经元中研究分子和电生理事件的能力将大大增强我们对潜在疾病机制的理解。发现诱导多能茎（IPS）细胞的发现，以及将它们分化为胶质，少突胶质细胞和各种神经元亚型的前景，为解决与验尸组织解释数据相关的问题提供了机会。我们将从对照受试者和患者患者获得的头发角质形成细胞中产生IPS细胞。患者队列将包括具有22q11删除的SZ的个体，这是一个遗传均匀的亚组。使用小鼠ES细胞中建立的方案将诱导的多能干细胞驱动以分化为谷氨酸能型金字塔神经元，并使用大量平行的深层测序（RNA-SEQ）进行表达分析，这比基于阵列杂交方法更敏感。将比较患者和对照组之间的表达谱，以鉴定SZ特异性差异。由于所使用的分化方案会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分析。提出的使用分化，患者特异性IPS细胞的实验将导致开发一种新方法来研究精神分裂症的潜在分子基础。 公共卫生相关性：这项研究描述了我们对精神分裂症患者不断增长的神经元的兴趣。神经元将是由所谓的诱导多能干细胞产生的，这些干细胞源自皮肤或发根。诱导的多能细胞具有发展为包括神经元在内的许多不同细胞类型的能力。研究精神分裂症患者的神经元将有助于我们对这种神秘疾病的理解，并可能导致新的疗法。