Testing cell autonomy of AD phenotypes using human IPS cells

使用人类 IPS 细胞测试 AD 表型的细胞自主性

• 批准号: 8461546
• 负责人: Lawrence S. Goldstein
• 金额: $ 21.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461546
• 关键词: Alleles; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein Precursor; Animal Model; Animals; Apolipoprotein E; Asses; Astrocytes; Axonal Transport; Biological Assay; C-terminal; Cell Communication; Cell Culture Techniques; Cell Cycle; Cell Line; Cell model; Cell physiology; Cells; Defect; Development; Diagnostic; Disease; Disease model; Failure; Genome; Human; Individual; Investigation; Lead; Light; Mediator of activation protein; Modeling; Neuroglia; Neurons; Older Population; Pathology; Pathway interactions; Patients; Peptide Fragments; Phenotype; Preclinical Drug Evaluation; Process; Protein Fragment; Proteins; Public Health; Relative (related person); Reporting; Signal Transduction; Staging; Synapses; System; Testing; Therapeutic; Toxic effect; Transcriptional Regulation; Vaccines; Work; apolipoprotein E-3; apolipoprotein E-4; base; disease phenotype; human disease; induced pluripotent stem cell; mixed cell culture; mutant; novel therapeutics; presenilin; relating to nervous system; research study; stem cell technology; Alleles; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein Precursor; Animal Model; Animals; Apolipoprotein E; Asses; Astrocytes; Axonal Transport; Biological Assay; C-terminal; Cell Communication; Cell Culture Techniques; Cell Cycle; Cell Line; Cell model; Cell physiology; Cells; Defect; Development; Diagnostic; Disease; Disease model; Failure; Genome; Human; Individual; Investigation; Lead; Light; Mediator of activation protein; Modeling; Neuroglia; Neurons; Older Population; Pathology; Pathway interactions; Patients; Peptide Fragments; Phenotype; Preclinical Drug Evaluation; Process; Protein Fragment; Proteins; Public Health; Relative (related person); Reporting; Signal Transduction; Staging; Synapses; System; Testing; Therapeutic; Toxic effect; Transcriptional Regulation; Vaccines; Work; apolipoprotein E-3; apolipoprotein E-4; base; disease phenotype; human disease; induced pluripotent stem cell; mixed cell culture; mutant; novel therapeutics; presenilin; relating to nervous system; research study; stem cell technology

DESCRIPTION (provided by applicant): Development of familial and sporadic Alzheimer's Disease (FAD and SAD) therapies requires deeper understanding of disease initiation and progression. A key unanswered question is whether all FAD and SAD neuronal misbehavior is solely the result of processes initiated or enhanced by secreted Amyloid Precursor Protein (APP) fragments such as A¿ and sAPP (and therefore cell non-autonomous), or whether A¿- independent intracellular processes driven by APP proteolytic products, e.g., the C-terminal fragment (CTF) (and therefore potentially cell autonomous) processes are a major contributor. Specifically, the major hypothesis for AD initiation and progression is the amyloid cascade hypothesis, which proposes that Ass peptide fragments of human APP are necessary and sufficient to initiate and to drive all downstream pathologies typical of AD progression including synaptic defects [1]. Alternative ideas include intracellular defects caused by presenilin or APP mutants/fragments that generate defects in endosomal or lysosomal pathways, axonal transport, neurotrophic signaling, transcriptional control, cell cycle reinitiation, oxidative defets, etc. [2-7]. There are also two-hit models in which A¿ is part of an initiating or enhancing insult n combination with intracellular insults [8-11]. These three types of models (autonomous, non-autonomous, two- hit) make distinct experimental predictions for mixed cell culture experiments using neurons derived from human induced pluripotent stem cells (hIPSC). For example, the (non-autonomous) amyloid cascade hypothesis, and related non-autonomous hypotheses based on toxicity of secreted fragments of APP (or other molecules) predict that mixtures of diseased and non-diseased neurons should cause disease phenotypes in the non-diseased neurons owing to secretion of toxic products by diseased neurons. Alternatively, in cell autonomous models of AD that do not posit secretion of toxic products, mixtures of diseased and non-diseased neurons should not lead to disease phenotypes in non-diseased neurons. Finally in two hit models, cell autonomous processes and cell nonautonomous processes might combine such that cell autonomous initiation of phenotypes might be enhanced by A¿ or other secreted toxic mediators. We propose to begin testing these ideas by further developing a new platform hIPSC human neuronal model of AD. These investigations, if successful, can shed new light on the relative contributions of autonomous and non-autonomous processes and two-hit models. We will use neurons made from hIPSC lines derived from a non-demented control patient (NDC), an FAD APPDp patient, and an SAD patient (called SAD2). We have two specific experimental aims: Aim 1) To test the hypothesis that some or all defects observed in purified neurons containing either an FAD APP duplication or a genome from an individual with SAD called SAD2 are cell-autonomous. Aim 2) To test the hypothesis that astrocytes carrying different ApoE alleles enhance or suppress AD phenotypes in purified neurons.

描述（由适用提供）：家庭和零星阿尔茨海默氏病的发展（FAD和SAD）疗法需要更深入地了解疾病的启动和进展。一个关键的未解决的问题是，所有时尚和悲伤的神经元不当行为是否仅仅是由分泌的淀粉样蛋白（APP）片段（如A和Sapp）（以及单元非自治）（因此是A€）启动或增强的过程的结果自主）过程是主要贡献者。具体而言，AD主动性和进展的主要假设是淀粉样蛋白级联假设，该假设提出，人类应用程序的屁股肽片段是必需的，足以启动并驱动所有典型的AD进展的下游病理，包括突触缺陷，包括突触缺陷[1]。另类思想包括由寄生虫或APP突变体/碎片引起的细胞内缺陷，这些缺陷会在内体或溶酶体途径中产生缺陷，轴突运输，神经营养信号传导，转录控制，细胞周期重新定性，氧化缺陷等[2-7]。还有两次打击的模型，其中A是与细胞内侮辱的侮辱或增强的一部分[8-11]。这三种类型的模型（自主，非自主，两次打击）对混合细胞培养实验进行了不同的实验预测，该模型使用源自人类诱导的多能干细胞（HIPSC）的神经元进行了混合细胞培养实验。例如，基于APP（或其他分子）分泌片段的毒性（或其他分子）的毒性，（非自主）淀粉样蛋白级联假设以及相关的非自治假设预测，由于伴有不良毒性的毒性神经元的毒性神经元的毒性混合物应引起疾病表型的疾病表型，从而导致疾病的疾病表型，从而使毒性不成熟的神经元伴有毒性的分泌。另外，在不阳性分泌有毒产品的AD细胞自主模型中，患病和未切除的神经元的混合物不应导致非疾病神经元的疾病表型。最后，在两个热门模型中，细胞自主过程和细胞非自主过程可能会结合，以使表型的细胞自主倡议可以通过A或其他分泌的有毒介质来增强。我们建议通过进一步开发一个新的平台HIPSC人类神经元模型来开始测试这些想法。这些研究，如果成功的话，可以新阐明自主和非自主过程和两次打击模型的相对贡献。我们将使用由源自非饮食对照患者（NDC），FAD APPDP患者和SAD患者（称为SAD2）的HIPSC系列的神经元。我们有两个具体的实验目的：目标1）检验以下假设：在纯化的神经元中观察到的某些或所有缺陷，其中包含fad应用程序复制或来自SAD的个体SAD2个体的基因组，称为SAD2是细胞自主。目的2）检验以下假设：携带不同APOE等位基因的星形胶质细胞增强或抑制纯化神经元中的AD表型。