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Phosphorylation-dependent human tau release

磷酸化依赖性人类 tau 释放

基本信息

批准号：
10046878
负责人：
DAEWOO LEE
金额：
$ 45.3万
依托单位：
OHIO UNIVERSITY ATHENS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10046878
关键词：
Affect Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease patient Alzheimer&apos s disease related dementia Alzheimer&apos s disease therapy Amino Acids Appearance Biological Assay Brain C-terminal CRISPR/Cas technology Cell Line Cells Data Disease Disease Progression Drosophila genus Enzyme-Linked Immunosorbent Assay Experimental Designs Foundations Goals Grant Human Human Cell Line Individual Interneurons Investigation Laboratories MAPT gene Mediating Mentors Methods Modeling Molecular Nature Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neurofibrillary Tangles Neurons Ohio Pathogenesis Pathogenicity Pathology Pattern Phosphorylation Phosphorylation Site Phosphotransferases Play Point Mutation Proline-Rich Domain Protein Kinase Proteins Protocols documentation Reproducibility Research Research Support Resources Role Seminal Site Small Interfering RNA Students Tauopathies Testing Transfection Transgenic Organisms Universities Western Blotting base experience experimental study graduate student hyperphosphorylated tau innovation insight interest knock-down mutant nerve stem cell neuronal excitability novel optogenetics prion-like tau Proteins tau phosphorylation tau-protein kinase therapeutic target training opportunity transmission process undergraduate student uptake

项目摘要

Neurofibrillary tangles (NFT) are a hallmark of Alzheimer's disease (AD) and related dementias. NFTs are of growing biomedical interest as the temporal and spatial pattern of NFT appearance in human brain correlates well with AD progression. The tangle is composed of microtubule associated protein tau, which can be hyper-phosphorylated and aggregated. Indeed, tau protein purified from the brains of AD patients is hyper-phosphorylated, which has led to investigations of the role of tau phosphorylation in mediating neurodegeneration. Alzheimer's pathogenic mechanisms still remain elusive. Recently, an intriguing concept of prion-like spreading of tau has emerged, which has the potential to transform AD research. A prion-like mechanism involving the transfer of hyper-phosphorylated tau between synaptically connected neurons underlies the seeding and spread of tau pathology. New insights into the molecular mechanisms of tau propagation will uncover potential therapeutic targets for slowing or even halting AD progression. Hyper-phosphorylation of tau is also known to be involved in tau release, causing its cell-to-cell propagation. However, there is still a significant gap in understanding how phosphorylation regulates tau release. We hypothesize that individual phosphorylation sites will differentially affect tau release and that studying this phenomena will uncover a currently unexplored network of endogenous protein kinases that act to regulate tau release by phosphorylating specific residues on tau. In particular, it is not known what phosphorylation sites of tau (pTau) are crucial for its release and kinases have yet to be identified for their role in tau release - a recognized seminal step in the prion-like spreading of tau protein. In Aim 1, we will study the role of specific phosphorylation sites of tau on its release. pTau sites to be examined will be primarily in proline-rich domain (PRD) and C-terminal region of tau as ~75% of disease associated pTau sites are found in these regions. In Aim 2, we will examine 5 kinases for their role in tau release as they are known to phosphorylate amino acids in PRD and C-terminal regions. Aim 3 will test if activity-driven endogenous hTau release is modulated by phosphorylation of tau at specific amino acids using a human neural cell line (ReNCell) endogenously expressing hTau. Our approach with Drosophila (Aims 1 & 2) and human neuronal culture models (Aim 3) will also provide excellent opportunities for training and mentoring both undergraduate and graduate students.

神经元缠结（NFT）是阿尔茨海默病（AD）和相关疾病的标志。痴呆症NFT作为NFT的时间和空间模式越来越受到生物医学的关注在人脑中的出现与AD进展密切相关。这团乱麻是由微管相关蛋白tau，其可以被过度磷酸化和聚集。事实上，从AD患者的脑中纯化的tau蛋白是高度磷酸化的，这使得导致研究tau磷酸化在介导神经变性中的作用。阿尔茨海默病的致病机制仍然是难以捉摸的。最近，一个有趣的概念， tau蛋白的朊病毒样扩散已经出现，这有可能改变AD研究。一朊病毒样机制涉及突触间过度磷酸化tau蛋白的转移连接的神经元是tau病理学的播种和传播的基础。新的见解的 tau蛋白传播的分子机制将揭示潜在的治疗靶点，甚至阻止AD的发展还已知tau的过度磷酸化参与tau释放，导致其细胞间传播。然而，在理解磷酸化是如何调节tau释放。我们假设，个别磷酸化位点将差异影响tau蛋白的释放，研究这种现象将揭示一个目前尚未探索的内源性蛋白激酶网络，其通过磷酸化调节tau蛋白释放 tau上的特定残基。特别是，尚不清楚tau蛋白（pTau）的磷酸化位点对于tau蛋白的释放至关重要，激酶在tau蛋白释放中的作用还有待鉴定，这是tau蛋白朊病毒样扩散中公认的开创性步骤。在目标1中，我们将研究tau蛋白的特异性磷酸化位点对其释放的作用。pTau 待检查的位点将主要在tau蛋白的富含脯氨酸的结构域（PRD）和C末端区域中因为约75%的疾病相关pTau位点发现于这些区域中。在目标2中，我们将检查5种激酶在tau释放中的作用，因为已知它们磷酸化氨基酸在PRD和C-末端区域。目的3将测试活性驱动的内源性hTau释放是否是通过使用人神经细胞系在特定氨基酸处磷酸化tau来调节（ReNCell）内源性表达hTau。我们对果蝇的方法（目标1和2），人类神经元培养模型（目标3）也将提供极好的培训机会并指导本科生和研究生。