Monitoring alterations of proteostasis in aging and Alzheimer's disease

监测衰老和阿尔茨海默病中蛋白质稳态的变化

• 批准号: 10294947
• 负责人: DANIELLE FAIVRE
• 金额: $ 4.08万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10294947
• 关键词: Affect; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease patient; Amino Acids; Amino Acyl Transfer RNA; Biochemistry; Biological; Biological Models; Brain; Brain region; Cells; Cerebrospinal Fluid; Collaborations; Complex; Data; Diet; Disease Progression; Environment; Equilibrium; Functional disorder; Future; Half-Life; Human; Impairment; Intercellular Fluid; Interdisciplinary Study; Leucine; Location; Mass Spectrum Analysis; Measures; Mentorship; Methods; Molecular Conformation; Monitor; Mus; Neurodegenerative Disorders; Patients; Peptide Hydrolases; Peptides; Phenotype; Protein Conformation; Protein Region; Proteins; Proteolysis; Proteome; Proteomics; Sampling; Stable Isotope Labeling; Structure; System; Techniques; Training; Universities; Washington; Work; aging brain; brain tissue; genome sciences; improved; insight; misfolded protein; prevent; protein aggregation; protein degradation; protein expression; protein folding; protein structure; proteostasis; statistics; tool; treatment response

Project Summary/Abstract Aging, Alzheimer’s disease (AD), and other neurodegenerative diseases have altered or impaired proteostasis. In healthy cells, proteostasis involves mechanisms for the stabilization of correctly folded proteins and the degradation of misfolded or damaged proteins. This balance maintains proteins in a specific conformation, concentration, and location to be functional, and the coordinated mechanisms prevent the accumulation of damaged proteins while permitting the continuous renewal of intracellular proteins. In aging, Alzheimer’s disease, and other neurodegenerative diseases, misfolded proteins aggregate into inclusions, which indicate that disruption of proteostasis has occurred. Effort has been put into determining what comprises the inclusions, but the cause of this altered or impaired proteostasis has never been determined. To improve our understanding of the causes of impaired proteostasis, we will examine the dynamic changes occurring in protein structure and protein turnover. We will use different types of samples and model systems to capitalize on the advantages of each. In SPECIFIC AIM 1, we propose to target proteins involved in AD in human cerebrospinal fluid with limited proteolysis to assess differences in protease accessibility where there were minimal changes in protein levels. By measuring changes in protease accessibility, we will gain insight into protein conformational changes. In SPECIFIC AIM 2, we will extend limited proteolysis to mouse brain to investigate differences in protease accessibility due to aging. In SPECIFIC AIM 3, we will examine protein turnover in 5 regions of mouse brain to reveal proteins with abnormal renewal related to aging. Measuring protein turnover requires the use of stable isotope labeling, which precludes the use of human samples. A benefit of using proteomic methods in this project is that we can examine many features at once and return to the data with new hypotheses in the future. The feasibility of the project is supported by the preliminary data generated for SPECIFIC AIM 1 and 2 and previous work done in the lab using the method proposed for SPECIFIC AIM 3. This proposal will be the first use of limited proteolysis-mass spectrometry with cerebrospinal fluid and brain tissue and the first example of protein turnover being studied in more than 2 regions of brain tissue. The results from both Aims will improve our understanding of proteostasis and deepen our understanding of the aging brain, Alzheimer’s disease, and other neurodegenerative diseases. This project will occur in a collaborative, interdisciplinary research environment in the University of Washington Genome Sciences department under the mentorship of Dr. Michael J. MacCoss, and the project will provide training in aging, Alzheimer’s disease, neurodegenerative diseases, statistics, and quantitative proteomics.

项目概要/摘要 衰老、阿尔茨海默病 (AD) 和其他神经退行性疾病已经改变或受损 在健康细胞中，蛋白质稳态涉及稳定正确折叠蛋白质的机制。 以及错误折叠或受损蛋白质的降解，这种平衡使蛋白质保持在特定的状态。 构象、浓度和位置发挥作用，协调机制可防止 受损蛋白质的积累，同时允许细胞内蛋白质的不断更新。 阿尔茨海默病和其他神经退行性疾病，错误折叠的蛋白质聚集成内含物， 这表明已经发生了蛋白质稳态的破坏。 包括内含物，但这种蛋白质稳态改变或受损的原因尚未确定。 为了提高我们对蛋白质稳态受损原因的理解，我们将检查动态变化 我们将使用不同类型的样品和模型系统。 为了充分利用各自的优势，在 SPECIFIC AIM 1 中，我们建议针对 AD 中涉及的蛋白质。 在人脑脊液中进行有限的蛋白水解，以评估蛋白酶可及性的差异 通过测量蛋白酶可及性的变化，我们发现蛋白质水平变化最小。 将深入了解蛋白质构象变化 在 SPECIFIC AIM 2 中，我们将扩展有限的蛋白水解。 在 SPECIFIC AIM 3 中，我们对小鼠大脑进行研究，以研究因衰老而导致的蛋白酶可及性差异。 将检查小鼠大脑 5 个区域的蛋白质更新，以揭示异常更新的蛋白质 与衰老相关的蛋白质周转需要使用稳定同位素标记，这排除了 在这个项目中使用蛋白质组学方法的一个好处是我们可以检查许多样本。 立即特征并在未来返回新假设的数据 该项目的可行性是。 得到为 SPECIFIC AIM 1 和 2 生成的初步数据以及之前在实验室完成的工作的支持 使用针对 SPECIFIC AIM 3 提出的方法。该提案将是有限蛋白水解质量的首次使用 脑脊液和脑组织的光谱测定以及正在研究的蛋白质周转的第一个例子 这两个目标的结果将提高我们对蛋白质稳态的理解。 加深我们对大脑老化、阿尔茨海默病和其他神经退行性疾病的了解。 该项目将在华盛顿大学的协作、跨学科研究环境中进行 基因组科学系在 Michael J. MacCoss 博士的指导下，该项目将提供 衰老、阿尔茨海默病、神经退行性疾病、统计学和定量蛋白质组学方面的培训。

项目成果

Sampling the proteome by emerging single-molecule and mass spectrometry methods.

通过新兴的单分子和质谱方法对蛋白质组进行采样。

• 发表时间: 2023-03
• 影响因子: 48
• 作者: MacCoss, Michael J;Alfaro, Javier Antonio;Faivre, Danielle A;Wu, Christine C;Wanunu, Meni;Slavov, Nikolai
• 通讯作者: Slavov, Nikolai