Exploring the Pathophysiology of AD and ADRDs with 3D Asteroid Models

使用 3D 小行星模型探索 AD 和 ADRD 的病理生理学

• 批准号: 10459727
• 负责人: Christine Cheng
• 金额: $ 91.62万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10459727
• 关键词: 3-Dimensional; Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease patient; Amyloid beta-Protein; Animal Model; Applications Grants; Astrocytes; Biological; Biology; Brain; Cell Culture Techniques; Cell Death; Cells; Complex; Data; Disease; Disease model; Dissociation; Electrophysiology (science); Elements; Exhibits; Exposure to; Functional disorder; Gene Expression; Genetic; Genetic Transcription; Glial Fibrillary Acidic Protein; Gliosis; Goals; Growth; HSF1; Heat shock proteins; Heat-Shock Proteins 90; Human; Immunohistochemistry; Label; Lead; Left; Link; Mediating; Microglia; Minor Planets; Modeling; Modification; Mus; Mutation; Nerve Degeneration; Neuroglia; Neuronal Differentiation; Neurons; Organoids; Pathologic; Pathology; Pathway interactions; Pattern; Physiological; Physiology; Process; Proteomics; Publications; RNA; Reporting; Research; Ribosomal Proteins; Ribosomes; Role; Seeds; Site; TP53 gene; Tauopathies; Technology; Transcript; Translations; Work; cell type; fluoro jade; induced pluripotent stem cell; inhibitor/antagonist; innovation; knock-down; neuropathology; novel; novel strategies; overexpression; protein complex; protein expression; resilience; response; single-cell RNA sequencing; stress granule; tau Proteins; tau aggregation; tau conformation; tau phosphorylation; tau-1; therapeutic target; thioflavine; three dimensional cell culture; transcription factor; transcriptome

This proposal presents a powerful new approach for modeling Alzheimer's Disease (AD) and AD-related diseases (ADRD), which enables discovery of important conceptual advances in our understanding of disease mechanisms. We use mixed cultures of human neurons and astrocytes developed from iPSCs from APP717V and isogenic control subjects. These are combined together into in 3D organoids, termed “asteroids”. Exposing the asteroids to propagated tau oligomers (oTau) causes the neurons develop tau pathology and neurodegeneration, creating a novel model for AD and ADRD, termed AstAD. The AstADs rapidly and progressively develop many components tauopathy, including oligomeric tau, phospho-tau, misfolded tau, fibrillar tau and neurodegeneration. Our studies with the AstAD model reveal that oTau disrupts ribosomal biology, selectively changing levels of key ribosomal proteins known to exert non-ribosomal functions directed at regulating the transcription factor HSF1, stress granules and the p53 pathway. These actions of oTau appear to induce of a harmful dysfunctional network of heat shock proteins (HSPs) and p53 mediated activation of cell death. However, selective dissociation of dysfunctional HSP complexes with the HSP90 inhibitor PU-H71, elicits a surprising, powerful reduction oTau pathology and neurodegeneration. Dysregulation of ribosomal protein levels has been observed in proteomic studies of AD, but the significance of ribosomal dysregulation has never been questioned. Our observations lead to the hypothesis that oTau dysregulates ribosomal biology in AstAD, producing deleterious patterns of ribosomal protein and HSP expression We will use the AstAD model to determine how ribosomal dysregulation induced by Aβ, oTau or Aβ/oTau affects neurons and astrocytes. We will also use the AstAD cultures to explore functions of neuron/glial interactions in disease, and determine how protective strategies directed against neurons, glia or both provide resilience. Aim 1 will determine the pattern of physiology, pathology and gene expression produced by Aβ, oTau, fibrillar tau or Aβ/oTau. We will examine the biology of ribosomes, HSPs and RNA translation in the AstAD model, as well as characterize key pathologies associated with AD and ADRD. Aim 2 will elucidate the complex biology of tauopathies among different types of human AD and ADRD cases by examining effects on the physiological, pathological and transcriptional responses in the AstAD cultures. We will induce neuropathology in AstAD by exposure to brain lysate, oTau or fibrillar tau produced from different types of tauopathies (AD, FTD-tau, PSP, CBD and PART) as well as comparison to FTD-TDP, which does not exhibit Aβ or tau pathology. Aim 3 will determine the roles of key ribosomal and HSPs in neurons or glia during the pathological response to Aβ and oTau. We will knockdown or over-express oTau-responsive ribosomal or HSP transcripts in neurons and/or astrocytes. We will then determine the patterns of physiological, pathological and transcriptional responses to these genetic modifications. This work will lead to novel mechanisms and therapeutic targets in AD and ADRD.

该提案提出了一种强大的新方法，用于建模阿尔茨海默病（AD）和AD相关的 疾病（ADRD），这使得我们能够在疾病的理解中发现重要的概念性进展 机制等我们使用从APP 717 V的iPSC开发的人神经元和星形胶质细胞的混合培养物， 和同基因对照组。这些结合在一起成为3D类器官，称为“小行星”。暴露 小行星到增殖的tau寡聚体（oTau）导致神经元发展tau病理学， 神经退行性疾病，创建了一种新的AD和ADRD模型，称为AstAD。AstAD迅速和 逐渐发展许多组分tau蛋白病，包括寡聚tau蛋白、磷酸化tau蛋白、错误折叠tau蛋白， 纤维状tau蛋白和神经变性。我们对AstAD模型的研究表明，oTau破坏了核糖体， 生物学，选择性地改变已知发挥非核糖体功能的关键核糖体蛋白质的水平， 在调节转录因子HSF 1、应激颗粒和p53通路方面。oTau的这些行为似乎 诱导热休克蛋白（HSPs）和p53介导的细胞活化的有害的功能失调网络 死亡然而，功能失调的HSP复合物与HSP 90抑制剂PU-H71的选择性解离， 这表明Tau病理和神经变性的令人惊讶的、有力的减少。核糖体调节异常 在AD的蛋白质组学研究中已经观察到蛋白质水平，但核糖体失调的意义 从未被质疑过我们的观察导致oTau失调核糖体生物学的假设 在AstAD中，产生核糖体蛋白和HSP表达的有害模式 模型，以确定Aβ、oTau或Aβ/oTau诱导的核糖体失调如何影响神经元， 星形胶质细胞我们还将使用AstAD培养物来探索神经元/神经胶质相互作用在疾病中的功能， 确定针对神经元、神经胶质或两者的保护策略如何提供弹性。目标1将 确定由Aβ、oTau、纤维状tau或 Aβ/aTau。我们还将研究AstAD模型中核糖体、热休克蛋白和RNA翻译的生物学特性 与AD和ADRD相关的主要病理特征。目的2将阐明复杂的生物学 不同类型的人AD和ADRD病例中的tau蛋白病， AstAD培养物中的病理和转录反应。我们将在AstAD中诱导神经病理学， 暴露于由不同类型的tau蛋白病（AD，FTD-tau，PSP， CBD和PART）以及与不表现出Aβ或tau病理的FTD-TDP的比较。目标3将 确定关键核糖体和热休克蛋白在神经元或神经胶质细胞对Aβ的病理反应中的作用， oTau。我们将敲低或过表达神经元中的oTau应答性核糖体或HSP转录物，和/或 星形胶质细胞。然后，我们将确定生理，病理和转录反应的模式， 这些基因改造。这项工作将导致新的机制和治疗AD和ADRD的目标。