Using induced pluripotent stem cells to identify cellular phenotypes of autism

使用诱导多能干细胞识别自闭症的细胞表型

• 批准号: 8314027
• 负责人: Ricardo E. Dolmetsch
• 金额: $ 79.2万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8314027
• 关键词: Autistic Disorder; Biological; Biological Assay; Cell Line; Cells; Defect; Development; Disease; Etiology; Fibroblasts; Fingerprint; Genes; Genetic; Genotype; Goals; Harvest; Lead; Link; Measures; Mental disorders; Microscope; Morphology; Mutation; Neurons; Patients; Phenotype; Skin; base; human stem cells; in vitro Assay; induced pluripotent stem cell; migration; patch clamp; stem cell technology

Psychiatric diseases are devastating and poorly understood. The goal of this project is to identify the cell biological phenotypes that underlie autism and other psychiatric disorders, taking advantage of new developments in human stem cell technology. I propose to harvest skin fibroblasts from patients with autism, to reprogram these cells to generate induced pluripotent stem (iPS) cells, and to differentiate the iPS cells into neurons. I will then use a set of semi-automated in vitro assays to develop a phenotypic fingerprint for each cell line focusing on the developmental and functional phenotypes that are likely to lead to autism. We will use automated microscopes, cell sorters and semi-automated patch clamps to measure the differentiation, migration, survival, morphology, and excitability of neurons derived from patients. Finally, we will take advantage of the genetic information available for some autistic patients to determine whether deletion, duplication, or mutation of specific genes leads the phenotypes observed in the neurons from that patient. This approach has the potential to revolutionize our study of autism and other psychiatric disorders. It will allow us to link the phenotype and genotype of patients with the cell biological defects that give rise to disease. In addition, it will allow us to develop cell-based assays to both investigate the etiology of the disease and to find new treatments for these untreatable disorders.

精神疾病具有毁灭性，但人们对其知之甚少。该项目的目标是 识别自闭症和其他精神疾病背后的细胞生物学表型 利用人类干细胞技术的新发展来治疗疾病。我 建议从自闭症患者身上采集皮肤成纤维细胞，对这些细胞进行重新编程 产生诱导多能干 (iPS) 细胞，并将 iPS 细胞分化为 神经元。然后，我将使用一组半自动体外测定来开发表型 每个细胞系的指纹，重点关注发育和功能表型 可能会导致自闭症。我们将使用自动显微镜、细胞分选仪和 半自动膜片钳测量分化、迁移、存活、 来自患者的神经元的形态和兴奋性。最后，我们将采取 利用一些自闭症患者可用的遗传信息来确定 特定基因的缺失、重复或突变是否会导致表型 在该患者的神经元中观察到。这种方法有可能 彻底改变我们对自闭症和其他精神疾病的研究。它将允许我们链接 具有细胞生物学缺陷的患者的表型和基因型 疾病。此外，它将使我们能够开发基于细胞的测定法来研究 该疾病的病因学并为这些无法治疗的疾病找到新的治疗方法。