iPSC Modeling of AD Using Progerin

项目总结/摘要 年龄是阿尔茨海默病（AD）的最大风险因素。然而，AD的细胞和动物模型未能 概括了人类衰老，未能捕捉疾病病理学的关键方面。我们的长期目标是 了解衰老如何影响AD发病机制。因此，我们建议开发一个诱导 基于多能干细胞（iPSC）的AD模型，包括加速老化，目的是更多 对AD的发病和进展进行稳健建模。最近的研究结果表明，核纤层蛋白A生物学的扰动 可能导致AD。在初步研究中，我们发现LMNA的显著增加， 编码核被膜蛋白核纤层蛋白A的ZMPSTE24， 在尸检证实的AD脑组织和AD脑的激光解剖神经元中， 与年龄匹配的对照组相比。我们预测，LMNA和ZMPSTE24水平的这些变化将导致 导致法尼基化的前核纤层蛋白A增加，这已被证明会加速衰老表型， 包括获得异常的核纤层，和核细胞质区室化的损伤， 类似于LMNA同种型早老蛋白的积累的染色质组织和基因表达。我们 假设前核纤层蛋白A或早老蛋白强制表达加速了年龄相关的变化和疾病 在源自AD患者的iPSC-皮质细胞中的表型。首先，我们将确定核纤层蛋白A是否 表达和加工在AD脑和AD易感的iPSC皮层细胞中受到干扰。然后我们将 确定强迫早老蛋白表达是否会导致AD易感iPSC中的衰老相关功能障碍， 皮质神经元和星形胶质细胞。最后，我们将定义和量化强制早老蛋白表达对 与AD病理学相关的细胞表型，包括iPSC-2D中的A β和tau分泌、聚集和转换 皮质细胞和3D前脑类器官。本研究的结果将确定是否存在扰动 核纤层蛋白A生物学与AD相关，并可能导致AD发病机制， 在人AD iPSC-皮质细胞模型中并入核纤层蛋白A相关衰老参数的新模型。这些 这些发现将为研究衰老对疾病机制的贡献开辟新的途径。 潜在的AD病理学，并将推进AD体外模型的开发，以帮助设计和 验证潜在的治疗策略。

Project Summary/Abstract Age is the strongest risk factor for Alzheimer's disease (AD). However, cell and animal models of AD fail to recapitulate human aging and fail to capture key aspects of disease pathology. Our long-term goal is to understand how aging contributes to AD pathogenesis. Therefore, we propose to develop an induced pluripotent stem cell (iPSC)-based model of AD that incorporates accelerated aging, with the objective of more robustly modeling AD onset and progression. Recent findings indicate that perturbations in lamin A biology may contribute to AD. In preliminary studies, we have found a significant increase in LMNA, the gene that encodes the nuclear envelope protein lamin A, and a significant decrease in ZMPSTE24, a prelamin A processing enzyme, in autopsy-confirmed AD brain tissue and in laser-dissected neurons from AD brains compared to age-matched controls. We predict that these changes in LMNA and ZMPSTE24 levels would cause an increase in farnesylated prelamin A, which has been shown to drive accelerated aging phenotypes, including acquisition of an abnormal nuclear lamina, and impairments in nucleocytoplasic compartmentation, chromatin organization and gene expression similar to the accumulation of the LMNA isoform progerin. We hypothesize that forced expression of prelamin A or progerin accelerates age-associated changes and disease phenotypes in iPSC-cortical cells derived from AD patients. First, we will determine whether lamin A expression and processing are perturbed in AD brains and AD-predisposed iPSC-cortical cells. We will then determine whether forced progerin expression causes aging-related dysfunction in AD-predisposed iPSC- cortical neurons and astrocytes. Finally, we will define and quantify the effects of forced progerin expression on cell phenotypes related to AD pathology, including Aβ and tau secretion, aggregation and turnover, in iPSC-2D cortical cells and in 3D forebrain organoids. The results from this study will determine whether perturbations in lamin A biology are associated with AD, and potentially contribute to AD pathogenesis, and will establish a novel model incorporating lamin A-related aging parameters in a human AD iPSC-cortical cell model. These findings will open novel avenues for investigating the contribution of aging to the disease mechanisms underlying AD pathology and will advance the development of in vitro models of AD to aid in the design and validation of potential therapeutic strategies.