Using induced pluripotent stem cells to identify cellular phenotypes of autism

精神疾病是毁灭性的，人们对此知之甚少。该项目的目标是 确定自闭症和其他精神疾病的细胞生物学表型 利用人类干细胞技术的新发展，我 我建议从自闭症患者身上收集皮肤成纤维细胞，重新编程这些细胞， 产生诱导多能干（iPS）细胞，并使iPS细胞分化为 神经元然后，我将使用一套半自动化的体外试验， 每种细胞系的指纹图谱，侧重于发育和功能表型 很可能导致自闭症。我们将使用自动显微镜，细胞分选仪， 半自动膜片钳测量细胞的分化、迁移、存活 形态学和源自患者的神经元的兴奋性。最后，我们将 一些自闭症患者可以利用遗传信息来确定 是否特定基因的缺失、重复或突变导致表型 在病人的神经元中观察到的。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。它能让我们把 具有引起以下疾病的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检测方法， 疾病的病因，并为这些无法治愈的疾病找到新的治疗方法。

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.