Define the effect of CLU SNP on the risk to Alzheimer's disease

项目摘要 阿尔茨海默病(AD)是老年人中最常见的痴呆症，目前还没有治愈方法 疾病。阿尔茨海默病发病的分子和细胞机制尚待阐明 为这种疾病开发有效的治疗方法。已经为AD研究和研究生成了许多小鼠模型 这些模型提供了重要的见解，以帮助我们了解疾病的病理基础。 然而，由于老鼠和人类神经细胞之间存在显著的物种差异，建立 需要人类疾病建模平台来补充动物模型中的研究，以更好地了解AD。 人类诱导多能干细胞(HiPSCs)已被广泛用于疾病模型的研究。 IPSC技术的发展。HiPSCs已被用于模拟AD的各个方面。因为HiPSCs 他们的衍生物被认为是年轻的表型，hPSC来源的细胞被用来模拟 公元早期事件。直接重编程是另一种类型的重编程，它将一种类型的体细胞 细胞转化为另一个细胞，而不需要经过涉及广泛的表观遗传修饰的IPSC阶段，因此 能够产生具有细胞衰老关键要素的人类细胞。因此，直接 来自患者体细胞的重新编程细胞将使我们能够对AD的年龄相关病理进行建模。 载脂蛋白E4是最强的遗传危险因素，CLU rs11136000 SNP的C等位基因是第三强的遗传危险因素 对于AD。这项建议的目的是定义CLU rs11136000 SNP单独或与 载脂蛋白E4对阿尔茨海默病的风险并揭示其潜在的分子和细胞机制 通过HiPSCs或通过直接重新编程生成的细胞模型。我们将使用基因编辑的同源细胞 明确CLU单核苷酸多态与载脂蛋白E亚型的联合作用。此外，由于HiPSC来源的细胞和 来自相同捐赠者的直接重编程细胞具有相同的遗传背景，但不同 细胞老化状态，它们代表了同基因的细胞平台，使我们能够专门研究这种影响 细胞老化的影响。这些等基因模型将使我们能够概括与年龄相关的表型并揭示 阿尔茨海默病的新病理机制。由于CLU和ApoE在星形胶质细胞中都有高表达， 我们建议使用星形胶质细胞单独或与载脂蛋白E一起定义CLU SNP的作用。我们假设 该CLU以ApoE异构体和年龄依赖的方式调节AD病理。因此，我们建议 具体目标：目标1：获得不同APOE/CLU基因和细胞衰老的星形胶质细胞 来自HiPSC或通过直接重新编程的状态。目标2：确定APOE和CLU变体对 应用星形胶质细胞-神经元或星形胶质细胞-OPC共培养研究AD的发病机制。目标3：确定 人脑APOE/CLU基因分型与增龄及基因表达的变化拟议的研究将会有所帮助。 为了确定CLU SNP在年龄相关AD病理发展中的作用，揭示 阿尔茨海默病的发病机制，并设计新的治疗策略。

Project Summary Alzheimer’s disease (AD) is the most common form of dementia in the elderly and there is no cure for this disease. The molecular and cellular mechanisms underlying AD pathogenesis remains to be elucidated to develop effective therapies for this disease. Many mouse models have been generated for AD research and these models provide important insights to aid our understanding of the pathological basis of the disease. However, because there are significant species differences between mouse and human neural cells, establishing human disease modeling platforms is needed to complement studies in animal models to better understand AD. Human induced pluripotent stem cells (hiPSCs) have been widely used for disease modeling since the development of the iPSC technology. hiPSCs have been used to model various aspects of AD. Because hiPSCs and their derivatives have been considered phenotypically young, hiPSC-derived cells have been used to model early events of AD. Direct reprogramming is another type of reprogramming that converts one type of somatic cells into another without going through the iPSC stage that involves extensive epigenetic modifications, thus enabling generation of human cells that possess key elements of cellular aging. Therefore, directly reprogrammed cells derived from patient somatic cells would allow us to model age-related pathologies of AD. ApoE4 is the strongest and the C allele of the CLU rs11136000 SNP is the third strongest genetic risk factor for AD. The objective of this proposal is to define the effect of the CLU rs11136000 SNP alone or together with ApoE4 on the risk to AD and uncover molecular and cellular mechanisms underlying the effect, using human cellular models generated from hiPSCs or through direct reprogramming. We will use gene-edited isogenic cells to define the effect of CLU SNP in combination with ApoE isoform. In addition, because hiPSC-derived cells and directly reprogrammed cells derived from the same donors have the same genetic background but different cellular aging status, they represent isogenic cellular platforms that will enable us to specifically study the effect of cellular aging. These isogenic models will allow us to recapitulate age-associated phenotypes and uncover novel pathological mechanisms underlying AD. Because both CLU and ApoE are highly expressed in astrocytes, we propose to define the effect of the CLU SNP alone or together with ApoE using astrocytes. We hypothesize that CLU modulates AD pathologies in an ApoE isoform- and age-dependent manner. Therefore, we propose following Specific Aims: Aim 1: To derive astrocytes with different APOE/CLU genotypes and cellular aging status from hiPSCs or through direct reprogramming. Aim 2: To define the effect of APOE and CLU variants on AD pathogenesis using astrocyte-neuron or astrocyte-OPC co-cultures. Aim 3: To determine the relationship of APOE/CLU genotypes and aging with gene expression change in human brains. The proposed studies will help to define the roles of the CLU SNP in the development of age-associated AD pathologies, to uncover mechanisms underlying AD pathogenesis, and to design novel therapeutic strategies for AD.