Functional Characterization of Tau Mutation and Post-translational Modifications

Tau 突变和翻译后修饰的功能表征

基本信息

批准号：
10572436
负责人：
Avi Jacob Samelson
金额：
$ 12.29万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-15 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10572436
关键词：
Alzheimer&apos s Disease American Animal Model Antibodies Behavior Biological Biological Process Biophysics Brain CRISPR interference Categories Cellular biology Characteristics Collaborations Complement Complex Deposition Development Disease Disease Progression Electron Transport Environment Etiology Event Functional disorder Genes Goals Human In Vitro Induced pluripotent stem cell derived neurons Kinetics Licensing Link Mass Spectrum Analysis Mentors Mitochondria Molecular Molecular Conformation Mutation Neurodegenerative Disorders Neurons Onset of illness Pathologic Peptide Hydrolases Pharmaceutical Preparations Phase Phosphorylation Positioning Attribute Post-Translational Protein Processing Postdoctoral Fellow Property Proteins Proteolysis Proteomics Research Research Personnel Scientist Spectrum Analysis Structure Symptoms Tauopathies Techniques Therapeutic Training Work career crosslink effective therapy functional genomics genome-wide in vivo innovation insight mutation screening new technology new therapeutic target novel novel therapeutic intervention protein aggregation protein misfolding skills tau Proteins tau aggregation tau interaction tau mutation tau-1 therapeutically effective

项目摘要

PROJECT SUMMARY/ ABSTRACT Protein aggregation is a hallmark of neurodegenerative diseases, including Alzheimer's Disease, and these diseases lack effective therapeutics. We currently lack an understanding of the molecular and cellular mechanisms controlling protein aggregation in the human brain, which would enable new therapeutic strategies. The protein tau aggregates in the brain in a number of neurodegenerative diseases called tauopathies, including Alzheimer's Disease. In early stages of disease, tau aggregates only in specific neurons despite being expressed in every neuron in the brain, implying that specific factors in the cellular environment predispose tau to aggregation. Similarly, tau mutations are associated with the onset of only specific tauopathies. Together, these disease features imply that tau is exquisitely sensitive to both its sequence properties and its cellular environment. Post-translational modifications (PTM) are a mechanism by which the cellular environment can act on a protein similarly to mutation. Tau is heavily post-translationally modified and changes to tau PTMs are correlated with progression of disease. Indeed, tau phosphorylation and proteolysis have been proposed to be central events in the onset and progression of tauopathies. Similarly, mutations are correlated with early disease onset and are known to hasten in vitro tau aggregation. Mutations can also cause changes in PTMs by changing tau interaction partners. The function and causality of these changes to tau sequence, however, is unknown. I hypothesize that PTMs and mutations license tau to access specific conformations to form aggregates. The goal of this proposal is to comprehensively identify (1) the biological basis of tau PTM changes and (2) how tau mutation and PTMs cause aggregation. I have shown that mitochondrial electron transport chain dysfunction causes a remodeling of tau PTMs, including the accumulation of a tau proteolytic fragment. In Aim 1, I will acquire new training in mass spectrometry (MS)-based proteomics to determine the tau PTM changes that occur due to ETC dysfunction and how those control tau aggregation. In Aim 2, I will use deep mutational scanning (DMS) to comprehensively probe tau's sequence-structure relationship. As part of Aim 2, I will use cross-link MS to directly compare in vitro and in vivo tau states to will reveal the structural mechanisms for the identified PTM and sequence changes. I am ideally positioned to complete the proposed research, as this proposal complements my training in protein biophysics and functional genomics in iPSC-derived neurons with novel MS-based techniques. Completion of this proposal will identify which PTMs and mutations control tau aggregation, as well as the development of new technologies to probe tau structure in vitro and in vivo. Completion of this proposal will provide me with essential skills and training to be a successful independent investigator.

项目摘要/摘要蛋白质聚集是神经退行性疾病的标志，包括阿尔茨海默氏病和这些疾病缺乏有效的治疗剂。我们目前对分子和控制人脑中蛋白质聚集的细胞机制，这将使新的治疗策略。大脑中蛋白质tau聚集在许多神经退行性疾病中，称为tauopathies，包括阿尔茨海默氏病。在疾病的早期，尽管在大脑的每个神经元中表达，这意味着细胞环境中的特定因素倾向于tau的聚集。同样，tau突变仅与特定的发作有关 tauopathies。这些疾病的特征共同表示Tau对其序列非常敏感性质及其细胞环境。翻译后修饰（PTM）是一种机制，细胞环境可以作用于蛋白质类似于突变。 tau经过重大翻译后修改，对tau PTM的变化是相关的随着疾病的进展。实际上，已经提出tau磷酸化和蛋白水解为中心 auopathies的发作和进展中的事件。同样，突变与早期疾病发作相关并已知会加速体外tau聚集。突变也会通过更改tau引起PTM的变化互动伙伴。但是，这些变化对tau序列的功能和因果关系尚不清楚。我假设PTM和突变许可TAU可以访问特定构象以形成聚集体。该提议的目的是全面识别（1）Tau PTM的生物学基础变化和（2）tau突变和PTM如何引起聚集。我已经证明了线粒体电子运输链功能障碍会导致tau PTM的重塑，包括tau蛋白水解的积累分段。在AIM 1中，我将获得基于质谱（MS）的蛋白质组学的新培训，以确定TAU PTM发生了由于ETC功能障碍而发生的变化以及那些如何控制Tau聚集。在AIM 2中，我将深入突变扫描（DMS），以全面探测Tau的序列关系。作为目标2的一部分，我将使用交联MS直接比较体外和体内tau状态将揭示结构机制对于已识别的PTM和序列变化。理想情况下，我的位置可以完成拟议的研究，因为该建议补充了我在IPSC衍生的神经元中蛋白质生物物理学和功能基因组学方面的培训采用新颖的基于MS的技术。该提案的完成将确定哪些PTM和突变控制 tau聚集以及在体外和体内探测tau结构的新技术的发展。该建议的完成将为我提供基本的技能和培训，以成为成功的独立研究者。