Programming age in iPS models of Alzheimer's disease

阿尔茨海默病 iPS 模型中的编程年龄

• 批准号: 9360837
• 负责人: LORENZ P. STUDER
• 金额: $ 43.2万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9360837
• 关键词: Address; Age; Aging; Alzheimer disease screening; Alzheimer' s Disease; Alzheimer' s disease model; Back; Biochemical; Biological Assay; CRISPR library; CRISPR screen; Cell Line; Cell model; Cells; Characteristics; Chronic; Dendrites; Disease; Ectopic Expression; Embryo; Epigenetic Process; Exhibits; Fetal Development; Fibroblasts; Genes; Genetic; Genetic Predisposition to Disease; Genetic Transcription; Goals; Herpes Simplex Infections; Heterochromatin; Hip region structure; Human; In Vitro; Late-Onset Disorder; Measures; Modeling; Mutation; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear; Nuclear Lamina; Parkinson Disease; Pathologic; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Population; Premature aging syndrome; Process; Progeria; Property; Proteins; Publishing; Reporting; Research; Risk Factors; Side; Syndrome; Tacrolimus Binding Proteins; Technology; Telomere Shortening; Testing; Tyrosine 3-Monooxygenase; age related; amyloid precursor protein processing; base; cell type; disease phenotype; dopaminergic neuron; early onset disorder; effective therapy; familial Alzheimer disease; fetal; genome-wide; human disease; human pluripotent stem cell; improved; induced pluripotent stem cell; innovation; insight; interest; knock-down; loss of function; mutant; neurodegenerative phenotype; neuromelanin; novel; novel strategies; pluripotency; presenilin-1; relating to nervous system; screening; tool

Project Summary Alzheimer’s disease (AD) is the most common neurodegenerative disorder in humans. Despite several decades of intense research there are currently still no effective treatments for AD. There has been considerable interest in exploring the use of induced pluripotent stem cells (iPSCs) as a novel tool to study AD, as the technology can capture the precise genetic background of a given AD patient. Furthermore, it is now possible to generate large numbers of such patient-specific, human iPSC-derived neurons on demand. This represents a powerful tool to study AD disease mechanisms in vitro, and to develop and test novel candidate therapies using iPSC-based screening assays. However, a problem that has plagued the iPSC field is the immature, fetal-like nature of the resulting neurons that does not match the age-related characteristics of AD patients. Furthermore, any age- related cellular markers that are present in primary cells from AD patients appear to be rejuvenated after reprogramming back to pluripotency. To address those limitations, we have recently reported on strategies to artificially trigger age-related marker expression in iPSC-derived neurons by manipulating pathways known to cause premature aging. Furthermore, we have provided proof-of-concept for using such induced aging strategies in iPSC models of Parkinson’s disease. Those “induced aging” strategies include the ectopic expression of progerin, a mutant form of the nuclear lamina protein LMNA, and the shortening of telomeres prior to and during neural differentiation. Additional candidate strategies reported by other labs include knockdown of the RanBP17 gene, a factor involved in nuclear/cytoplasmic transport and global loss of heterochromatin, an epigenetic cellular change that triggers premature aging-like feature in fibroblast. The goal of the current study is to test current and to develop novel induced aging strategies that may be particularly suitable to model AD. First, we aim to compare several current induced aging strategies in AD-iPSC-derived cortical neurons to assess, side-by-side, their ability to trigger age-related marker expression and AD-related biochemical and degenerative changes. To model the AD-specific effects, we will use isogenic AD-iPSC lines, recently established, carrying mutations in APP(Swe) and presenilin 1 (PSEN1(M146V). Second, we will identify and validate novel candidate induced aging strategies of particular relevance in triggering degenerative phenotypes in AD- but not in control iPSC-derived neurons. Third, we will use the most promising strategies from Aim 1 and Aim 2 to determine their ability to trigger disease phenotypes in iPSC-derived cortical neurons derived from patients with sporadic AD. Sporadic AD patients represent the most common form of the disease and have been particularly difficult to model using conventional iPSC technology. The proposed study will address whether current or novel induced aging strategies can contribute to improved iPSC models of AD. Similar approaches may apply to many other late-onset disorders and could offer fundamental insights into the mechanism of neuronal aging.

项目摘要 阿尔茨海默氏病（AD）是人类最常见的神经退行性疾病。尽管几十年 在激烈的研究中，目前仍然没有有效的AD治疗方法。有很大的兴趣 在探索诱导的多能干细胞（IPSC）作为研究AD的新工具时，技术可以 捕获给定的AD患者的精确遗传背景。此外，现在可以生成大的 这种特定于患者的人IPSC衍生的神经元的数量按需。这代表了一个强大的工具 在体外研究AD疾病机制，并使用基于IPSC的新候选疗法开发和测试新型候选疗法 筛选分析。但是，困扰IPSC领域的问题是未成熟的胎儿样本 导致的神经元与AD患者的年龄相关特征不符。此外，任何年龄 - AD患者的原代细胞中存在的相关细胞标记似乎在 重新编程回多能。为了解决这些限制，我们最近报告了 通过操纵已知的途径，在IPSC衍生的神经元中人为触发与年龄相关的标记表达 导致过早衰老。此外，我们提供了使用此类诱发衰老策略的概念验证 在帕金森氏病的IPSC模型中。那些“诱发的衰老”策略包括 Progerin，一种核薄片蛋白LMNA的突变形式，以及端粒的缩短之前和期间 神经分化。其他实验室报告的其他候选策略包括敲除RANBP17 基因，涉及核/细胞质转运的因素和异染色质的全球丧失，一种表观遗传细胞 更改该触发的成纤维细胞中衰老的过早特征。当前研究的目的是测试电流和 开发新型诱导的衰老策略，这些策略可能特别适合于建模AD。首先，我们的目的是比较 AD-IPSC衍生的皮质神经元中的几种当前诱导的衰老策略，以并排评估其能力 触发与年龄相关的标记表达以及与广告相关的生化和退化性变化。建模 AD特异性效应，我们将使用最近确定的ISEOGENIC AD-IPSC线，在App（SWE）中携带突变（SWE） 和呈现1（PSEN1（M146V））。其次，我们将识别并验证新型候选人诱导的衰老策略 与触发AD型的退化表型特别相关，而不是控制IPSC衍生的神经元。 第三，我们将使用AIM 1中最有前途的策略并确定其触发疾病的能力 IPSC衍生的皮质神经元的表型来自零星AD患者。零星的广告患者 代表了疾病的最常见形式，并且很难使用常规 IPSC技术。 拟议的研究将解决当前或新型诱导的衰老策略是否有助于 改进了AD的IPSC模型。类似的方法可能适用于许多其他晚期疾病，可以提供 对神经元衰老机制的基本见解。