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的同工型改变了患阿尔茨海默氏病（AD）或心血管疾病的风险，并且是 也与特殊的寿命有关。具体而言，E2/E2基因型与异常 E4等位基因与寿命负相关的寿命。 APOE基因的E4变体也是一个 AD的主要危险因素，与大脑中AB沉积较高有关。相应地， APOE E2等位基因与AD相关神经变性的风险较低有关。机制调制 与E3和E4基因型相比，E2介导的延长寿命尚不清楚。一个假设是 APOE E2等位基因是神经保护性的，并补偿了被折叠率诱导的神经元功能障碍 蛋白质表达在衰老和疾病中。此外，APOE E2等位基因的发展计划 可能与ApoE E4等位基因不同。这是基于暗示apoe的文献的基础 基因型影响大脑结构和基因表达开始产前发育并继续延迟 生活。我们提出的研究将利用由ISEOGEN性诱导的多能干细胞（IPSC）。 CRISPR/CAS9表达APOE蛋白的三种同工型（E2，E3和E4）。使用基因工程 初步研究，我们在对照IPSC和 亨廷顿疾病-HD-IPSC。干细胞研究的最新进展表明IPSC可能会提供新颖 衰老和疾病的模型。我们将使用干细胞模型调查特殊寿命的作用 衰老和疾病中的因子APOE2具有以下特定目的：特定目的1。我们将表征 使用系统的E2/E2，E3/E3和E4/E4基因型的等源IPSC的细胞和功能差异 生物学方法。特定目的2：确定促进寿命的APOE变体是否会增强压力 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。 特定目标3。我们将确定APOE2的表达或APOE细胞产生的因子是否增加了 老年小鼠的健康跨度。总体而言，我们的方法协同确定了许多独特的方法来确定 与阿尔茨海默氏病相关的自然发生的遗传变异如何调节多个的健康跨度 组织和潜在的寿命。成功完成我们提出的研究可能会揭示与 与年龄相关疾病和衰老的潜在治疗剥削。