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）是由神经递质传递缺陷引起的。SZ中的aminamatergic通路的分析已经解决了使用各种不同的实验方法，其中之一是使用阵列杂交进行死后脑组织或外周血白细胞的基因表达谱。这些研究为SZ的发病机制提供了许多令人兴奋的线索。然而，当白细胞被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来自尸检标本的数据。此外，这些生物材料来源不能用于电生理分析。因此，在培养的患者特异性神经元中研究分子和电生理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，以及驱使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患者获得的发根角质形成细胞产生iPS细胞。患者队列将包括具有22 q11缺失的SZ个体，这是一个遗传同质亚组。诱导的多能干细胞将使用在小鼠ES细胞中建立的方案被驱动分化成神经元能锥体神经元，并且表达谱将使用大规模平行深度测序（RNA-seq）进行，其比基于阵列杂交的方法更敏感。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前体锥体神经元，因此也将进行电生理学分析。使用分化的患者特异性iPS细胞的拟议实验将导致开发一种研究精神分裂症潜在分子基础的新方法。 公共卫生相关性：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。神经元将由所谓的诱导多能干细胞产生，这些干细胞来自皮肤或发根。诱导多能细胞具有发育成许多不同细胞类型的能力，包括神经元。研究精神分裂症患者的神经元将有助于我们理解这种神秘的疾病，并可能导致新的治疗方法。