The Influence of Tau Post-Translational Modifications on the Propagation of Tau Pathology in Alzheimer's Disease

Tau 翻译后修饰对阿尔茨海默氏病 Tau 病理学传播的影响

• 批准号: 10676180
• 负责人: JOHN R DICKSON
• 金额: $ 17.28万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-04 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676180
• 关键词: 3-Dimensional; Acetylation; Address; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease patient; Amino Acids; Biological Assay; Biology; Biosensor; Brain region; Cell Culture Techniques; Cell model; Cell physiology; Cells; Charge; Chemicals; Clinical; Communication; Cytoplasm; Data; Deacetylation; Dementia; Development; Development Plans; Disease; Disease Progression; Endosomes; Evaluation; Flow Cytometry; Fluorescence Resonance Energy Transfer; Functional disorder; Galectin 3; Grant; Human; Impaired cognition; In Vitro; Induced pluripotent stem cell derived neurons; K-Series Research Career Programs; Literature; Luciferases; Manuscripts; Mentors; Mentorship; Modeling; Modification; Molecular; Molecular Weight; Mutation; Neuroanatomy; Neurofibrillary Tangles; Neurons; Pathologic; Pathology; Pattern; Performance; Phosphorylation; Physicians; Play; Positron-Emission Tomography; Post-Translational Protein Processing; Preparation; Process; Property; Protein Dephosphorylation; Publishing; Radiology Specialty; Recombinants; Research; Role; Scientist; Stains; Stereotyping; Techniques; Testing; Writing; abeta accumulation; brain cell; career; career development; defined contribution; design; endosome membrane; experience; experimental analysis; experimental study; extracellular; immunocytochemistry; innovation; insight; machine learning algorithm; meetings; mutant; neuronal circuitry; neuropathology; novel therapeutic intervention; protein structure prediction; skills; tau Proteins; tau aggregation; tau mutation; tau-1

Project Summary/Narrative This K08 Mentored Clinical Scientist Research Career Development Award application investigates the role of tau post-translational modifications on the cellular processes involved in the stereotyped spreading of tau pathology that occurs as Alzheimer’s disease progresses. The cellular mechanisms underlying this process include the cell-to-cell propagation of pathologic tau from a donor cell to a recipient cell, escape from the endosomal compartment in the recipient cell, and subsequent seeding of tau aggregation in the cytoplasm. Previous studies published in the literature, as well as preliminary data related to this project, suggest that tau post-translational modifications, especially phosphorylations and acetylations, influence these processes. The overall hypothesis of the project is that tau post-translational modifications found on pathologic tau in Alzheimer’s disease influence the cellular mechanisms involved in the spreading of tau pathology. The specific aims of the project focus on investigating the impact of disease-associated tau post-translational modifications on 1) the cell-to-cell propagation of tau, 2) the endosomal escape of tau, and 3) the seeding activity of tau. The research strategy for this project uses human induced pluripotent stem cell-derived neurons to study the processes outlined in the specific aims, and mutations mimicking post-translational modifications (pseudophosphorylations and pseudoacetylations) will be used to assess the contribution of defined combinations of specific residues. Specific Aim 1 uses an expression construct with the wildtype or mutant tau sequences to identify donor and recipient cells by flow cytometry to assess cell-to-cell propagation. Specific Aim 2 uses treatment of neurons with wildtype or mutant recombinant tau proteins and evaluation of endosomal membrane integrity and tau endosomal escape using immunocytochemistry with galectin-3 staining and a split luciferase assay, respectively. Specific Aim 3 uses tau Förster resonance energy transfer (FRET) biosensor cell models to detect tau seeding activity of the wildtype or mutant recombinant tau proteins or Alzheimer’s disease-derived tau preparations with or without in vitro dephosphorylation and/or deacetylation. The career development plan of this proposal is designed to prepare the applicant for independence as a scientist. The applicant will take courses that provide a conceptual framework for design, performance, and analysis of the experiments described above. Scientific communication skills will be further developed by coursework and experience in writing manuscripts/grants and presenting the results at scientific meetings. The applicant had assembled a team of mentors, both the primary mentor and Scientific Advisory Board, to assist in the applicant’s development over the course of the project. The combination of the research experience, career development activities, and mentorship the applicant receives is designed to prepare the applicant for a successful independent career as a physician-scientist in dementia research.

项目摘要/叙述 该K08指导的临床科学家研究职业发展奖应用程序调查了 TAU翻译后修饰在与刻板印象扩散有关的细胞过程中的作用 随着阿尔茨海默氏病的发展而发生的tau病理。该过程的基础机制 包括从供体细胞到受体细胞的病理tau的细胞间传播，逃脱 受体细胞中的内体隔室，以及随后在细胞质中tau聚集的播种。 先前发表在文献上的研究以及与该项目有关的初步数据表明Tau 翻译后修饰，尤其是磷酸化和乙酰化，会影响这些过程。 该项目的总体假设是TAU在病理学上发现的翻译后修饰 阿尔茨海默氏病中的tau会影响tau病理传播所涉及的细胞机制。 该项目的具体目的专注于研究与疾病相关后翻译的影响 修改1）tau的细胞间传播，2）tau的内体逃生，3）播种 tau的活动。该项目的研究策略使用人类诱导的多能干细胞衍生 神经元研究特定目的中概述的过程，并模仿翻译后的突变 修改（假磷酸化和假乙酰化）将用于评估 特定残差的定义组合。特定目标1使用野生型或 突变的tau序列通过流式细胞仪来鉴定供体和受体细胞，以评估细胞间传播。 特定的目标2使用野生型或突变重组tau蛋白的神经元处理以及评估 内体膜完整性和TAU内体逃逸，使用乳糖素3染色的免疫细胞化学 和分裂的荧光素酶测定法。特定目标3使用tauförster共振能量传递（FRET） 生物传感器细胞模型检测野生型或突变体重组tau蛋白或 阿尔茨海默氏病衍生的tau制剂，有或没有体外去磷酸化和/或脱乙酰化。 该提案的职业发展计划旨在为申请人准备独立 作为科学家。申请人将参加为设计，性能，性能提供概念框架的课程 以及上述实验的分析。科学沟通技巧将由 课程工作和写作手稿/赠款的经验，并在科学会议上介绍结果。这 申请人召集了一个主要导师和科学顾问委员会的导师团队，以协助 在该应用程序过程中的开发中。研究经验的结合， 申请人所接受的职业发展活动以及概念训练旨在为申请人做好准备 作为痴呆症研究中的身体科学家的成功独立职业。