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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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.

项目概要 ApoE 亚型可降低患阿尔茨海默病 (AD) 或心血管疾病的风险，并且还与超凡的长寿相关。具体而言，e2/e2 基因型与超凡的寿命相关。长寿，而 e4 等位基因与长寿呈负相关。 ApoE 基因的 e4 变体也是一个。 AD 的主要危险因素，并且与大脑中较高水平的抗体沉积相关。 ApoE e2 等位基因与 AD 相关神经变性的风险降低有关。与 e3 和 e4 基因型相比，e2 介导的寿命延长尚不清楚。 ApoE e2 等位基因具有神经保护作用，可补偿错误折叠引起的神经元功能障碍此外，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表明 ApoE 的文献。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我们提出的研究将利用经过改造的同基因诱导多能干细胞（iPSC）。使用基因工程 CRISPR/Cas9 表达 ApoE 蛋白的三种亚型（E2、E3 和 E4）。初步研究，我们在对照 iPSC 中生成了携带 e2/e2、e3/e3 和 e4/e4 基因型的系，亨廷顿病-HD-iPSC。干细胞研究的最新进展表明，iPSC 可能提供新的治疗方法。我们将利用干细胞模型研究长寿的作用。 ApoE2 在衰老和疾病中发挥着重要作用，具体目标如下：具体目标 1. 我们将描述使用系统分析具有 e2/e2、e3/e3 和 e4/e4 基因型的同基因 iPSC 的细胞和功能差异具体目标 2：确定促进长寿的 ApoE 变异是否会增强压力。抵抗力和存活率，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 ApoE2 或 ApoE 细胞产生的因子的表达是否会增加总体而言，我们的方法结合了多种独特的方法来确定。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健康寿命组织和潜在的寿命的成功完成我们提出的研究可能会揭示相关机制。年龄相关疾病和衰老的潜在治疗开发。