Human induced neuronal stem cell models of familial Alzheimer's disease

描述（由申请人提供）：我们最近描述了将来自未受影响个体和阿尔茨海默病（AD）患者的人皮肤成纤维细胞定向转化为CNS神经元表型，称为人诱导神经元细胞（hiN）1。在此，我们建议进一步开发hiN细胞作为AD建模的工具，并随后在更详细的AD细胞机制分析中验证该方法。拟议研究的重点是家族性AD（FAD）相关疾病与早老素-1（PSEN 1）或早老素-2（PSEN 2）的明确遗传病变。对具有确定突变的FAD进行这种分析的明显优点是，这有利于遗传“拯救”研究以及功能的遗传解剖。然而，最终，hiN细胞技术最令人兴奋的方面可能是它可以允许对“散发性”疾病进行细胞分析。在这项工作中要测试的首要假设是，FAD病理生理学的细胞方面是：（1）细胞自主神经元，并通过表观遗传重编程维持，因此适合hiN细胞建模。(2)包括在索马和突触处改变的胞内囊泡运输。本研究的主要成果是：（1）建立新型的人类神经元细胞模型，用于AD机制的研究和药物筛选。(2)定向重编程工具，可广泛应用于神经系统疾病的研究。 公共卫生相关性：我们描述了一种通过人类皮肤细胞的定向转化来产生人类神经元的新方法。当这些神经元来源于家族性阿尔茨海默氏症患者时，会显示出典型的疾病病理变化。在这里，我们建议调查人类阿尔茨海默病患者神经元细胞和分子缺陷的机制。

DESCRIPTION (provided by applicant): We recently described the directed conversion of human skin fibroblasts from unaffected individuals and Alzheimer's disease (AD) patients to a CNS neuron phenotype, termed human induced neuronal cells (hiN) 1. Herein we propose to further develop hiN cells as tools for AD modeling, and subsequently to validate the approach in a more detailed analysis of cellular mechanisms of AD. The focus of the proposed studies is on familial AD (FAD)-associated disease with defined genetic lesions in presenilin-1 (PSEN1) or presenilin-2 (PSEN2). A clear advantage of such an analysis of FAD with defined mutations is that this facilitates genetic 'rescue' studies, as well as genetic dissection of function. Yet ultimately, perhaps the most exciting aspect of the hiN cell technology is that it may permit a cellular analysis of 'sporadic' disease. The overarching hypotheses to be tested in this work are that cellular aspects of FAD pathophysiology are: (1) Cell-autonomous to neurons and maintained through epigenetic reprogramming, and therefore amenable to hiN cell modeling. (2) Include altered intracellular vesicular trafficking at the soma and the synapse. The proposed deliverables for this proposed work are: (1) Novel human neuronal cell models for dissection of AD mechanisms and drug screening. (2) Directed reprogramming tools that may be broadly applied to the study of neurological disease. PUBLIC HEALTH RELEVANCE: We describe a novel approach to the generation of human neurons by the directed conversion of human skin cells. Such neurons, when derived from patients with familial forms of Alzheimer's, show pathological changes that typify the disease. Here we propose to investigate the mechanism by which human Alzheimer patient neurons develop cellular and molecular deficits.