Resilience pathways modeling human longevity-promoting ApoE variants in induced pluripotent stem cells

项目总结 ApoE的亚型会降低患阿尔茨海默病(AD)或心血管疾病的风险，而且 也与非凡的长寿有关。具体地说，e2/e2基因与特殊的 而e4等位基因与长寿呈负相关。载脂蛋白E基因的e4变体也是一种 阿尔茨海默病的主要危险因素，与大脑中较高水平的抗体沉积有关。相应地， APOE e2等位基因与AD相关神经变性的风险较低。调节机制 E2与e3和e4基因相比是否延长寿命尚不清楚。一种假设是， ApoE e2等位基因对错误折叠所致神经元功能障碍的神经保护作用 蛋白质在衰老和疾病中的表达。此外，ApoE e2等位基因的发育程序 可能与载脂蛋白E e4等位基因不同。这是基于一系列文献，这些文献表明载脂蛋白E 基因分型影响大脑结构和基因表达，从产前发育开始一直持续到晚期 生活。我们建议的研究将利用同基因诱导多能干细胞(IPSCs)， CRISPR/Cas9表达载脂蛋白E的三种异构体(E2、E3和E4)。利用基因工程在 初步研究，我们在对照IPSCs中获得了携带e2/e2、e3/e3和e4/e4基因的系 亨廷顿病-HD-iPSCs。干细胞研究的最新进展表明，IPSCs可能提供新的 衰老和疾病的模型。我们将使用干细胞模型研究异常长寿的作用 因子APOE2在衰老和疾病中具有以下特定目的：特定目的1。我们将表征 应用A系统研究e2/e2、e3/e3和e4/e4三种等位基因IPSCs的细胞和功能差异 生物学方法。具体目标2：确定促进长寿的载脂蛋白E变体是否会增加压力 并确定与各种变异体的神经保护作用相关的途径。 具体目的3.我们将确定APOE2的表达或ApoE细胞产生的因子是否增加 老龄小鼠的健康寿命。总体而言，我们的方法结合了许多独特的方法来确定 与阿尔茨海默病相关的自然发生的基因变异如何在多个 组织和潜在的寿命。我们建议的研究的成功完成可能揭示 对年龄相关疾病和衰老的潜在治疗开发。

PROJECT SUMMARY Isoforms of ApoE modify the risk for developing Alzheimer’s disease (AD) or cardiovascular disease, and are also associated with exceptional longevity. Specifically, the e2/e2 genotype is associated with exceptional longevity while the e4 allele is negatively associated with longevity. The e4 variant of the ApoE gene is also a major risk factor for AD and is associated with higher levels of Ab deposition in the brain. Correspondingly, the ApoE e2 allele is associated with a lower risk of AD-related neurodegeneration. The mechanisms modulating extended lifespan mediated by e2 compared to e3 and e4 genotypes are not clear. One hypothesis is that the ApoE e2 allele is neuroprotective and compensates for neuronal dysfunction induced by misfolded protein expression in aging and disease. In addition, the developmental program for the ApoE e2 allele may be distinct from the ApoE e4 allele. This is based on a body of literature that suggests the ApoE genotypes effect brain structure and gene expression beginning prenatal development and continuing into late life. Our proposed studies will utilize isogenic induced pluripotent stem cells (iPSCs) engineered with CRISPR/Cas9 to express the three isoforms of the ApoE protein (E2, E3 and E4). Using genetic engineering in preliminary studies, we generated lines carrying e2/e2, e3/e3 and e4/e4 genotypes in control iPSCs and Huntington’s Disease-HD-iPSCs. Recent advances in stem cell research suggest that iPSCs may provide novel models of aging and diseases. We will investigate using stem cell models the role of the exceptional longevity factor ApoE2 in aging and disease with the following Specific Aims: Specific Aim 1. We will characterize the cellular and functional differences in isogenic iPSCs with e2/e2, e3/e3 and e4/e4 genotypes using a systems biology approach. Specific Aim 2: To determine whether longevity-promoting ApoE variants enhance stress resistance and survival and identify the pathways relevant to the neuroprotective effects of the various variants. Specific Aim 3. We will determine if expression of ApoE2 or factors produced by ApoE cells provide increased health span in aged mice. Overall, our approach synergizes a number of unique methodologies to determine how naturally occurring genetic variants associated with Alzheimer’s disease modulate health span in multiple tissues and potentially lifespan. Successful completion of our proposed studies may reveal mechanisms with potential therapeutic exploitation for age-related disease and aging.