Linking tau proteostasis with neuronal activity in FTD

将 tau 蛋白稳态与 FTD 中的神经元活动联系起来

• 批准号: 9524981
• 负责人: Li Gan
• 金额: $ 1.45万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9524981
• 关键词: Address; Advisory Committees; Affect; Alzheimer' s Disease; Animal Model; Autophagocytosis; Brain; CRISPR interference; Candidate Disease Gene; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Communities; Complex; Cultured Cells; Data; Data Analyses; Development; Disease; Ensure; Experimental Designs; Failure; Feedback; Frontotemporal Lobar Degenerations; Gene Expression; Generations; Heterogeneity; Human; In Vitro; Libraries; Link; Mediating; Mutation; Nerve Degeneration; Neuronal Dysfunction; Neurons; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Post-Translational Protein Processing; Process; Proteins; Proteomics; Reagent; Research; Research Infrastructure; Resources; Role; Sampling; Shapes; Structure; Tauopathies; Technology; Testing; Toxic effect; United States National Institutes of Health; Validation; Vision; Work; data integration; data sharing; experience; extracellular; functional outcomes; genome-wide; human data; human tissue; hyperphosphorylated tau; in vivo; induced pluripotent stem cell; innovative technologies; insight; member; multi-electrode arrays; mutant; novel; novel therapeutics; patch clamp; proteostasis; screening; synaptic function; synergism; tau Proteins; tau mutation; transcriptome; uptake

ABSTRACT — OVERALL COMPONENT Neurodegenerative tauopathies, including Alzheimer's disease (AD) and frontotemporal lobar degeneration with tau inclusions (FTLD-tau), are characterized by tau inclusions of hyperphosphorylated tau. However, which tau species are pathogenic and how they induce neuronal dysfunction remain elusive. We hypothesize that there is a crosstalk between tau proteostasis imbalance, in the form of intracellular accumulation and intercellular spread, and neuronal dysfunction. Our vision statement is: Linking tau proteostasis with neuronal activity in FTD. We propose to use a combination of unbiased discovery and focused hypothesis- driven approaches to dissect the mechanistic connection. The Center, composed of three projects (P1–P3) and five cores (MS, CRISPR, Human, Data and Admin), aims at addressing the following fundamental questions in tau pathogenesis. In Specific Aim 1, we will address what causes tau to accumulate and spread in FTD. P1 and P3 will work together with MS core to dissect whether aberrant post-translational modifications (PTMs) are critically involved in modulating tau's failure to be degraded and subsequent release. P1, P2 and P3 will work together with CRISPR core to dissect the uptake and seeding mechanisms, using a combination of hypothesis-driven candidate approaches and genome-wide CRISPRi/a library screening. P1, P2 and P3 will work together with the Human core to validate the findings from iPSC neurons in human tissues. In Specific Aim 2, we will determine how tau proteostasis imbalance induces neuronal dysfunction. Despite large amount of evidence supporting the accumulation and spread of tau pathology in animal models and in cultured cells, little is known about the mechanisms behind this toxicity and their direct impact on neuronal/synaptic function in human neurons. P1 and P2 will work together to define the effects of FTD mutations on human neuronal activity both at the single-cell and network levels using whole-cell patch clamp and multi-electrode array (MEA), respectively. P3 will work with P1 and P2 to examine if altered autophagy affects neuronal activity. P1, P2 and the CRISPR core will work together to examine the functional effects of tau oligomerization. In Specific Aim 3, we will examine how neuronal activity modulates tau proteostasis. Both in vitro and in vivo studies showed that the release of tau is activity-dependent, supporting a feedback mechanism by which aberrant neuronal activity further alters tau proteostasis. P1 will work with the MS and CRISPR cores to dissect the mechanisms underlying the activity-dependent tau release. P1 and P2 will work synergistically to address if and how neuronal activity could affect the uptake (P2) or seeding (P1) of pathogenic tau. P3 will work with P1 and P2 to determine how altering neuronal activity modulates different autophagic pathways. In summary, the Center is strategically structured to maximize conceptual and technological synergies, full data integration and cross-validation. We anticipate discovery of novel insights into mechanisms underlying tau toxicity, generation of novel resources and reagents, and development of innovative technology platforms.

摘要-整体组件 神经退行性tau蛋白病，包括阿尔茨海默病（AD）和额颞叶变性 具有tau内含物（FTLD-tau），其特征在于过度磷酸化tau的tau内含物。然而，在这方面， 哪些tau种类是致病性的以及它们如何诱导神经元功能障碍仍然是难以捉摸的。我们假设 在tau蛋白抑制失衡之间存在串扰，以细胞内积累的形式， 细胞间扩散和神经元功能障碍。我们的愿景声明是：将tau蛋白酶抑制与 FTD中的神经元活性。我们建议结合无偏见的发现和集中的假设- 驱动的方法来剖析机械连接。该中心由三个项目组成（P1-P3） 和五个核心（MS，CRISPR，Human，Data和Admin），旨在解决以下基本问题 tau发病机制的问题。在具体目标1中，我们将讨论导致tau积累和扩散的原因 在FTD中。P1和P3将与MS核心一起工作，以分析是否存在异常的翻译后修饰 （PTM）关键地参与调节tau的降解失败和随后的释放。P1、P2和 P3将与CRISPR核心一起工作，使用组合来剖析吸收和播种机制， 假设驱动的候选方法和全基因组CRISPRi/a文库筛选。P1、P2和P3将 与Human core合作，验证人类组织中iPSC神经元的发现。在特定 目的2，我们将确定tau蛋白稳态失衡如何诱导神经元功能障碍。尽管数额巨大 支持tau病理学在动物模型和培养细胞中积累和传播的证据， 关于这种毒性背后的机制及其对神经元/突触功能的直接影响知之甚少 在人类神经元中。P1和P2将共同确定FTD突变对人神经元的影响 使用全细胞膜片钳和多电极阵列在单细胞和网络水平上的活性 (MEA)，分别。P3将与P1和P2一起检查改变的自噬是否影响神经元活动。P1， P2和CRISPR核心将共同研究tau寡聚化的功能效应。在特定 目的3，我们将研究神经元活动如何调节tau蛋白稳态。体外和体内研究 表明tau蛋白的释放是活性依赖性的，支持一种反馈机制， 神经元活性进一步改变tau蛋白质稳态。P1将与MS和CRISPR核心合作， 活性依赖性tau释放的潜在机制。P1和P2将协同工作， 以及神经元活动如何影响致病性tau的摄取（P2）或接种（P1）。P3将与P1一起工作 和P2来确定改变神经元活性如何调节不同的自噬途径。总而言之， 中心的战略结构是为了最大限度地发挥概念和技术协同作用，全面的数据集成， 交叉验证我们期待着发现新的见解的机制，潜在的tau毒性，产生 新资源和试剂，以及创新技术平台的开发。