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

诱导多能干细胞中模拟人类长寿 ApoE 变异的弹性途径

• 批准号: 10417069
• 负责人: Lisa M Ellerby
• 金额: $ 87.96万
• 依托单位: BUCK INSTITUTE FOR RESEARCH ON AGING
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10417069
• 关键词: Aging; Alleles; Alzheimer associated neurodegeneration; Alzheimer' s Disease; Alzheimer' s disease risk; Amino Acids; Apolipoprotein E; Arginine; Astrocytes; Behavioral Symptoms; Bioinformatics; Biological Markers; Brain; CRISPR/Cas technology; Cardiovascular Diseases; Cells; Cholesterol; Coculture Techniques; Code; Cysteine; Data Set; Deposition; Development; Disease; E protein; Elderly; Fat-Soluble Vitamin; Gene Expression; Genes; Genetic Engineering; Genotype; Human; Huntington Disease; Lipoproteins; Literature; Longevity; Maintenance; Mediating; Methodology; Microglia; Modeling; Molecular Analysis; Monitor; Mus; Neuraxis; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Pathogenesis; Pathology; Pathway interactions; Phenotype; Play; Positioning Attribute; Process; Protein Isoforms; Proteins; Proteomics; Publications; Resistance; Risk; Signal Pathway; Stem Cell Research; Stress; Structure; Synaptic Membranes; Synaptic plasticity; System; Systems Biology; Testing; Therapeutic; Tissues; Variant; Work; age related; aged; apolipoprotein E-3; apolipoprotein E-4; base; engineered stem cells; genetic variant; genome editing; healthspan; human model; induced pluripotent stem cell; knock-down; misfolded protein; mutant; neuronal growth; novel; overexpression; prenatal; programs; protein expression; proteotoxicity; repaired; resilience; role model; single cell analysis; single-cell RNA sequencing; stem cell model; therapeutic target; transcriptomics

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.

项目摘要 ApoE的同种型改变了发展阿尔茨海默病（AD）或心血管疾病的风险，并且 也与长寿有关。具体来说，e2/e2基因型与异常的 而e4等位基因与寿命呈负相关。ApoE基因的e4变异体也是一种 是AD的主要危险因素，并与大脑中较高水平的Ab沉积相关。相应地 ApoE e2等位基因与AD相关神经退行性变风险降低相关。调节机制 与e3和e4基因型相比，由e2介导的寿命延长尚不清楚。一种假设是， ApoE e2等位基因具有神经保护作用并补偿错误折叠诱导的神经元功能障碍 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ApoE e4等位基因不同。这是基于一个机构的文献表明，载脂蛋白E 基因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。 生活我们提出的研究将利用经基因工程改造的同基因诱导多能干细胞（iPSC）， CRISPR/Cas9表达ApoE蛋白的三种同种型（E2、E3和E4）。利用基因工程 在初步研究中，我们在对照iPSC中产生了携带e2/e2、e3/e3和e4/e4基因型的细胞系， 亨廷顿病-HD-iPSCs。干细胞研究的最新进展表明，iPSC可以提供新的 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ApoE 2在衰老和疾病中的作用，具体目标如下：具体目标1。我们将描述 使用系统在具有e2/e2、e3/e3和e4/e4基因型的同基因iPSC中的细胞和功能差异 生物学方法。具体目标2：确定促进长寿的ApoE变体是否会增强应激 抗性和存活，并确定与各种变体的神经保护作用相关的途径。 具体目标3。我们将确定ApoE 2或ApoE细胞产生的因子的表达是否提供了增加 老年小鼠健康寿命。总的来说，我们的方法协同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遗传变异如何调节多种疾病中的健康跨度 组织和潜在的寿命。成功完成我们提出的研究可能会揭示机制， 对年龄相关疾病和衰老的潜在治疗开发。