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 这K 08指导临床科学家研究职业发展奖申请调查 tau蛋白翻译后修饰对参与细胞定型扩散的细胞过程的作用 随着阿尔茨海默病的进展而发生的tau病理学。这一过程背后的细胞机制 包括病理性tau从供体细胞到受体细胞的细胞间传播，从供体细胞逃逸， 受体细胞中的内体隔室，以及随后在细胞质中接种tau聚集。 先前发表在文献中的研究，以及与本项目相关的初步数据表明，tau蛋白 翻译后修饰，特别是磷酸化和乙酰化，影响这些过程。 该项目的总体假设是，在病理组织中发现的tau蛋白翻译后修饰， 阿尔茨海默病中的tau影响涉及tau病理学扩散的细胞机制。的 该项目的具体目标集中在研究疾病相关的tau蛋白翻译后的影响， 修饰1）tau的细胞间增殖，2）tau的内体逃逸，和3）接种 Tau的活动。该项目的研究策略使用人类诱导多能干细胞衍生的 神经元研究的过程中概述的具体目标，和突变模仿翻译后 修饰（假磷酸化和假乙酰化）将用于评估 特定残基的定义组合。特异性目的1使用具有野生型或 突变的tau序列，以通过流式细胞术鉴定供体和受体细胞，从而评估细胞到细胞的增殖。 具体目标2使用野生型或突变重组tau蛋白处理神经元，并评估神经元的功能。 利用半乳糖凝集素-3染色的免疫细胞化学研究内体膜完整性和tau内体逃逸 和裂解荧光素酶测定。具体目标3使用tau Förster共振能量转移（FRET） 生物传感器细胞模型，以检测野生型或突变重组tau蛋白的tau接种活性，或 具有或不具有体外脱磷酸化和/或脱乙酰化的阿尔茨海默病衍生的tau制剂。 本建议书的职业发展计划旨在为申请人的独立做好准备 作为一个科学家申请人将参加提供设计，性能， 和上述实验的分析。科学沟通技能将进一步发展， 在撰写手稿/赠款和在科学会议上介绍成果方面的课程和经验。的 申请人已经组建了一个导师团队，包括主要导师和科学顾问委员会，以协助 申请人在项目过程中的发展。结合研究经验， 职业发展活动，和指导，申请人收到的目的是准备申请人 作为痴呆症研究的医生科学家，成功的独立职业生涯。