Novel strategies for induction of aging in human iPSC-derived lineages towards improved models of late-onset diseases

诱导人类 iPSC 衍生谱系衰老的新策略，以改进迟发性疾病模型

• 批准号: 9924425
• 负责人: Doron Betel
• 金额: $ 56.85万
• 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: 2017-09-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9924425
• 关键词: Age; Aging; Astrocytes; Biochemical; Biological Assay; Brain; Cell Aging; Cell Line; Cell Lineage; Cell model; Cell physiology; Cells; Characteristics; Chromatin; Coculture Techniques; DNA Methylation; DNA Modification Methylases; DNA analysis; Data; Dendrites; Disease; Disease model; Dopamine; Ectopic Expression; Embryo; Epigenetic Process; Fibroblasts; Gene Expression Profiling; Genetic; Genomics; Goals; Herpes Simplex Infections; Human; In Vitro; Knowledge; LRRK2 gene; Lamin Type A; Late-Onset Disorder; Lead; Link; Malignant Neoplasms; Methodology; Midbrain structure; Modeling; Molecular; Molecular Genetics; Molecular Profiling; Monitor; Morphologic artifacts; Morphology; Mus; Mutation; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurologic; Neurons; Nuclear Lamina; Parkinson Disease; Pathologic; Pathology; Pathway interactions; Patients; Pharmacology; Phenotype; Physiological; Pluripotent Stem Cells; Premature aging syndrome; Process; Progeria; Proliferating; Proteins; Protocols documentation; Publishing; Rejuvenation; Reporter; Risk Factors; Sampling; Site-Directed Mutagenesis; Syndrome; Tacrolimus Binding Proteins; Technology; Transplantation; Tyrosine 3-Monooxygenase; Xenograft procedure; age related; aged; alpha synuclein; base; brain tissue; cell age; cell behavior; cell type; design; disease phenotype; dopaminergic neuron; drug discovery; early onset disorder; epigenetic profiling; experimental study; fascinate; gene therapy; human disease; human model; improved; in vivo; in vivo Model; induced pluripotent stem cell; insight; mouse model; mutant; nerve stem cell; neurodegenerative phenotype; neuromelanin; novel; novel strategies; stem cell model; tool; transcriptome sequencing

Project Summary The primary risk factor for most neurodegenerative diseases and many other human ailments such as cancer is old age. A major challenge in studying late-onset diseases is the accurate representation of the aging context in both in vitro and in vivo models of the disease. This is due in part to a lack of understanding of the cellular and molecular characteristics of the aging process. We previously defined a set of cellular changes associated with aging in cells from old donors and demonstrated that these age-related hallmarks are restored to a youthful state through reprogramming into induced pluripotent stem cells (iPSC) and maintained in such “rejuvenated” state upon re-differentiation into iPSC-derived cells. This phenomenon, while fascinating from a scientific perspective, also represents a concrete barrier for the use of iPSC for studying age-dependent disorders. Our group demonstrated that these aging characteristics can be reintroduced into iPSC-derived cells through simple expression of progerin, a mutant form of the lamin A (LMNA) protein that is responsible for the premature aging disorder Hutchinson-Gilford Progeria. However, inducing cellular age using a disease-causing factor may not be a faithful representation of physiological aging and potentially lead to pathological artifacts in modeling aging-related diseases. It is therefore necessary to develop a cellular model that will accurately reproduce the physiological aging context. Our hypothesis, supported by promising preliminary results, is that cellular aging is promoted by specific genetic and epigenetic changes that can be utilized to trigger an aged cellular state in models of late-onset disease. Here, we propose to develop a new approach for modeling age in iPSC-derived neuronal lineages in three specific steps. First, we aim to generate a comprehensive characterization of genetic and epigenetic features of aged cells using primary fibroblasts and brain tissues from young and old donors, which will provide genomic aging signatures of different cell lineages. We will then monitor those signatures during reprogramming and identify candidate determinants of cellular age. We will validate these signatures by targeted experiments as well as on independent samples. Second, using a combination of our previously identified cellular hallmarks of aging in conjunction with the newly identified genetic and epigenetic aging markers we will design optimal strategies to induce cellular aging. Third, we will use these strategies to study the impact of cellular age on the progression and pathology of Parkinson disease (PD) using iPSC-derived dopamine neurons from patients with genetic forms of PD. These will both be used for in vitro studies characterizing cellular PD manifestations as well as in vivo upon transplantation into PD mouse models to assess the impact of induced aging on cellular behavior and function in vivo. The results from this study will provide a more comprehensive understanding of the genetic and epigenetic changes that underlie human aging and importantly, provide a methodology to induce aging context in iPSC models of human diseases.

项目摘要 大多数神经退行性疾病和许多其他人类疾病（如癌症）的主要风险因素 就是年老研究迟发性疾病的一个主要挑战是准确地表示衰老 在疾病的体外和体内模型中的背景。这部分是由于缺乏对 衰老过程的细胞和分子特征。我们之前定义了一组细胞变化 与来自老年供体的细胞老化相关，并证明这些与年龄相关的标志可以恢复， 通过重新编程为诱导多能干细胞（iPSC）并维持在这种状态， 在再分化成iPSC衍生的细胞时，细胞处于“再生”状态。这种现象，虽然从一个迷人的 从科学的角度来看，也代表了使用iPSC研究年龄依赖性的具体障碍。 紊乱我们的研究小组证明，这些衰老特征可以重新引入iPSC衍生的细胞， 通过简单表达早老蛋白，一种突变形式的核纤层蛋白A（LMNA）蛋白，负责 早老症Hutchinson-Gilford早老症然而，使用致病的 因素可能不是生理老化的忠实代表，并可能导致病理性伪影， 模拟衰老相关疾病。因此，有必要开发一种细胞模型， 重现生理老化背景。我们的假设得到了初步结果的支持，即 细胞衰老是由特定的遗传和表观遗传变化促进的，这些变化可以用来触发衰老。 迟发性疾病模型中的细胞状态。在这里，我们建议开发一种新的方法来建模年龄 在iPSC衍生的神经元谱系中的三个具体步骤。首先，我们的目标是产生一个全面的 使用原代成纤维细胞和脑组织表征老化细胞的遗传和表观遗传特征 这将提供不同细胞谱系的基因组衰老特征。然后我们将 在重编程过程中监测这些特征，并确定细胞年龄的候选决定因素。我们将 通过有针对性的实验以及独立样本验证这些特征。第二，使用 结合我们以前发现的衰老细胞标志和新发现的 遗传和表观遗传衰老标记，我们将设计最佳策略来诱导细胞衰老。三是 使用这些策略来研究细胞年龄对帕金森病的进展和病理学的影响 (PD)使用来自遗传性PD患者的iPSC衍生的多巴胺神经元。这两个都将用于 用于表征细胞PD表现的体外研究以及移植到PD中后的体内研究 小鼠模型，以评估诱导衰老对体内细胞行为和功能的影响。 这项研究的结果将提供一个更全面的了解遗传和表观遗传 这些变化是人类衰老的基础，重要的是，提供了一种方法来诱导iPSC中的衰老背景。 人类疾病的模型。