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: 9904310
• 负责人: LORENZ P. STUDER
• 金额: $ 44.9万
• 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/9904310
• 关键词: Address; Age; Aging; Alzheimer disease screening; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Back; Biochemical; Biological Assay; CRISPR library; CRISPR screen; Cell Line; 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 model; 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; stem cell model; stem cell technology; 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.

项目概要 尽管已有几十年的历史，阿尔茨海默病（AD）仍然是人类最常见的神经退行性疾病。 经过深入的研究，目前仍然没有有效的治疗 AD 的方法。 探索使用诱导多能干细胞 (iPSC) 作为研究 AD 的新工具，因为该技术可以 此外，现在可以生成大量的 AD 患者的精确遗传背景。 按需提供此类患者特异性、人类 iPSC 衍生神经元的数量，这是一种强大的工具。 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 然而，困扰 iPSC 领域的一个问题是其不成熟、类似胎儿的性质。 由此产生的神经元与 AD 患者的年龄相关特征不匹配。 AD 患者原代细胞中存在的相关细胞标记物似乎在治疗后恢复了活力。 为了解决这些限制，我们最近报告了一些策略。 通过操纵已知的途径，在 iPSC 衍生的神经元中人为地表达与年龄触发相关的标记物 此外，我们还提供了使用这种诱导衰老策略的概念验证。 在帕金森病的 iPSC 模型中，这些“诱导衰老”策略包括异位表达。 早老蛋白（核纤层蛋白 LMNA 的一种突变形式）以及端粒在手术前和手术中的缩短 其他实验室报告的其他候选策略包括敲除 RanBP17。 基因，参与核/细胞质运输和异染色质整体丢失的因素，异染色质是一种表观遗传细胞 触发成纤维细胞过早衰老特征的变化当前研究的目标是测试当前和 开发可能特别适合 AD 模型的新型诱导衰老策略 首先，我们的目标是进行比较。 AD-iPSC 衍生的皮质神经元中目前的几种诱导衰老策略，以并行评估它们的能力 触发与年龄相关的标记物表达以及与 AD 相关的生化和退行性变化。 AD 特异性效应，我们将使用最近建立的同基因 AD-iPSC 系，其携带 APP(Swe) 突变 和早老素 1 (PSEN1(M146V))。其次，我们将确定并验证新的候选诱导衰老策略 与触发 AD 退行性表型特别相关，但与对照 iPSC 衍生神经元无关。 第三，我们将使用目标 1 和目标 2 中最有前途的策略来确定它们引发疾病的能力 来自散发性 AD 患者的 iPSC 来源的皮质神经元的表型。 代表了该疾病最常见的形式，并且使用传统方法建模特别困难 iPSC 技术。 拟议的研究将解决当前或新型诱导衰老策略是否有助于 改进的 AD iPSC 模型可能适用于许多其他迟发性疾病，并且可以提供类似的方法。 对神经衰老机制的基本见解。