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 此 K08 指导临床科学家研究职业发展奖申请调查了 tau 翻译后修饰对参与定型传播的细胞过程的作用 阿尔茨海默病进展时发生的 tau 蛋白病理学。该过程背后的细胞机制 包括病理性 tau 蛋白从供体细胞到受体细胞的细胞间传播、从 受体细胞中的内体区室，随后将 tau 蛋白聚集在细胞质中。 之前发表在文献中的研究以及与该项目相关的初步数据表明，tau 翻译后修饰，尤其是磷酸化和乙酰化，影响这些过程。 该项目的总体假设是，病理学上发现的 tau 翻译后修饰 阿尔茨海默病中的 tau 蛋白会影响 tau 蛋白病理学传播的细胞机制。这 该项目的具体目标侧重于调查与疾病相关的 tau 翻译后的影响 1) tau 的细胞间传播、2) tau 的内体逃逸和 3) 接种的修改 tau 的活性。该项目的研究策略使用人类诱导多能干细胞 神经元研究特定目标中概述的过程以及模仿翻译后的突变 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。具体目标 1 使用具有野生型或 突变 tau 序列，通过流式细胞术识别供体和受体细胞，以评估细胞间的增殖。 具体目标 2 使用野生型或突变型重组 tau 蛋白治疗神经元并评估 使用半乳糖凝集素 3 染色免疫细胞化学检测内体膜完整性和 tau 内体逃逸 和分裂荧光素酶测定，分别。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，用于检测野生型或突变型重组 tau 蛋白的 tau 接种活性，或 阿尔茨海默氏病衍生的 tau 制剂，具有或不具有体外去磷酸化和/或去乙酰化作用。 本提案的职业发展计划旨在帮助申请人为独立做好准备 作为一名科学家。申请人将参加提供设计、性能、概念框架的课程。 以及上述实验的分析。科学沟通能力将得到进一步发展 撰写手稿/资助以及在科学会议上展示结果的课程作业和经验。这 申请人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， 申请人接受的职业发展活动和指导旨在帮助申请人做好准备 作为痴呆症研究领域的医师科学家，取得了成功的独立职业生涯。