Neurons from pluripotent stem cells derived from schizophrenia patient fibroblast

描述（由申请人提供）：精神分裂症是影响约0.5%人口的主要公共卫生负担。该病的病因、发病机制和病理生理尚不清楚。目前的治疗方法只能帮助部分患者，而且有严重的副作用。在此，我们建议利用干细胞技术的重大突破，即从成纤维细胞等体细胞中产生多能干细胞的能力，来开发新的细胞工具，以帮助找到更好的治疗精神分裂症的方法。我们假设，从精神分裂症患者的成纤维细胞中分化出来的多能干细胞的神经细胞将表现出与精神分裂症死后大脑中检测到的病理和免疫病理结果相似的异常。为了验证这一假设，我们组建了一个在精神分裂症、干细胞、神经元发育和分化以及神经精神疾病细胞模型方面具有专业知识的团队。在目标1中，我们将仔细选择8名精神分裂症患者和8名匹配的对照组，并对他们进行皮肤活检。这些人都同意参加广泛的诊断访谈，以及其他涉及神经影像学、基因分析和神经心理学测试的协议。在Aim 2中，我们将从皮肤活检中生成成纤维细胞系，然后使用含有人类Oc3/4、Sox2、Klf4和c-Myc的逆转录病毒从成纤维细胞中生成多能干细胞。细胞的多能性将得到明确的确立，细胞将分化为神经元。在Aim 3中，我们将描述神经元的特征，并比较来自患者和对照组的细胞的特性。我们将强调神经炎形态和突触标记，特别是谷氨酸能和gaba能功能的指标。我们从这项研究中获得的试点数据将使我们能够竞争资金来扩大这项事业，有更多的患者和更系统的分化神经元检查。总之，概括精神分裂症病理方面的细胞系将作为开发精神分裂症治疗新靶点的有价值的工具。这些细胞系也可能被证明是筛选新治疗药物有效性的平台。公众健康启示：我们的提案旨在从精神分裂症患者的皮肤细胞中产生干细胞和随后的神经元，并确定这些细胞与从健康个体获得的细胞有何不同。这些细胞将提供一个有价值的工具来提高我们对精神分裂症生物学的理解，从而发现新的治疗靶点，从而可以设计药物或其他形式的治疗。这些细胞也可能被证明是直接有用的平台，用于筛选现有的化合物，作为可能的精神分裂症治疗剂。

DESCRIPTION (provided by applicant): Schizophrenia is a major public health burden affecting about 0.5% of the population. The etiology, pathogenesis, and pathophysiology of the disease is poorly understood. Current treatments help only some affected individuals and have serious side effects.Here we propose to take advantage of a major breakthrough in stem cell technology, the capacity to generate pluripotent stem cells from somatic cells such as fibroblasts, to develop novel cellular tools to help find better treatments for schizophrenia. We hypothesize that neuronal cells differentiated from pluripotent stem cells that have been derived from fibroblasts of schizophrenia patients will demonstrate abnormalities similar to the pathological and immunopathological findings detected in schizophrenia postmortem brain. To test this hypothesis, we have assembled a team with expertise in schizophrenia, stem cells, neuronal development and differentiation, and cell models of neuropsychiatric disease. In aim 1, we will carefully select and characterize 8 patients with schizophrenia and 8 matched controls, and obtain skin biopsies from them. These individuals will all have agreed to participate in extensive diagnostic interviews, as well as other protocols involving neuroimaging, genetic analysis, and neuropsychological testing. In Aim 2, we will generate fibroblast cell lines form the skin biopsies, and then use retroviruses containing human Oc3/4, Sox2, Klf4, and c-Myc to generate pluripotent stem cells from the fibroblasts. The pluripotency of the cells will be definitively established, and the cells will be differentiated into neurons. In Aim 3, we will characterize the neurons and compare the properties of cells derived from patients and from controls. We will emphasize neuritic morphology and synaptic markers, and particularly indicators of glutamatergic and GABAergic functions. The pilot data we obtain from this study will enable us to compete for funds to enlarge this enterprise, with more patients and a more systematic examination of differentiated neurons. Overall, cell lines that recapitulate aspects of the pathology of schizophrenia will serve as valuable tools in the development of new therapeutic targets for schizophrenia treatment. The cell lines may also prove of value as platforms for screening the effectiveness of new therapeutic agen. PUBLICE HEALTH REVELANCE: Our proposal is designed to generate stem cells, and subsequently neurons, from skin cells of individuals with schizophrenia, and to determine how these cells are different from cells obtained from healthy individuals. The cells will provide a valuable tool to improve our understanding of the biology of schizophrenia, and thereby of detecting new therapeutic targets for which drugs or other forms of treatment can be designed. The cells may also prove directly useful as platforms for screening existing compounds as possible therapeutic agents for schizophrenia.