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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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（由申请人提供）：家族性和散发性阿尔茨海默氏病（FAD 和 SAD）疗法的开发需要对疾病的发生和进展有更深入的了解，一个尚未解答的关键问题是所有 FAD 和 SAD 神经不良行为是否仅仅是启动或增强过程的结果。由分泌的淀粉样前体蛋白 (APP) 片段如 A¿和 sAPP （因此细胞是非自主的），或者是否 A¿ - 由 APP 蛋白水解产物驱动的独立细胞内过程，例如 C 末端片段 (CTF)（因此可能是细胞自主）过程是主要的具体贡献者。AD 起始和进展的主要假设是淀粉样蛋白级联假说，该假说提出人类 APP 的 Ass 肽片段对于启动和驱动 AD 进展的所有典型下游病理（包括突触缺陷）是必要且充分的[1]。早老素或 APP 突变体/片段会在内体或溶酶体途径、轴突运输、神经营养信号传导、转录控制、细胞周期重新启动、氧化缺陷等方面产生缺陷。 [2-7] 也有 A¿是与细胞内损伤相结合的起始或增强损伤的一部分[8-11]这三种类型的模型（自主、非自主、两次打击）对使用源自人类诱导的神经元的混合细胞培养实验做出了不同的实验预测。例如，（非自主）淀粉样蛋白级联假说，以及基于 APP（或）分泌片段毒性的相关非自主假说。其他分子）预测，由于患病神经元分泌有毒产物，患病和未患病神经元的混合物应在未患病神经元中引起疾病表型，或者，在不分泌有毒产物的 AD 细胞自主模型中，患病和非患病神经元的混合物不应导致非患病神经元出现疾病表型。最后，在两个命中模型中，细胞自主过程和细胞非自主过程可能会结合起来，从而可以通过以下方式增强细胞自主启动表型。一个我们建议通过进一步开发 AD 的新平台 hIPSC 人类神经模型来开始测试这些想法，如果成功，这些研究可以为自主和非自主过程的相对贡献提供新的线索。我们将使用来自非痴呆对照患者 (NDC)、FAD APPDp 患者和 SAD 患者（称为 SAD2）的 hIPSC 系制成的神经元。我们有两个具体的实验目标： 1) 检验在含有 FAD APP 重复或来自 SAD 个体（称为 SAD2）的基因组的纯化神经元中观察到的部分或所有缺陷是细胞自主的假设目标 2) 检验携带不同 ApoE 等位基因的星形胶质细胞增强的假设。或抑制纯化神经元中的 AD 表型。