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Analysis of protein interactions in neurodegenerative disease

神经退行性疾病中蛋白质相互作用的分析

基本信息

批准号：
10613978
负责人：
John R Yates III
金额：
$ 70.69万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10613978
关键词：
Affect Age Alzheimer like pathology Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease risk Alzheimer&apos s disease therapy Amino Acids Animal Disease Models Animal Model Antibodies Autophagocytosis Biochemical Biological Brain Brain Diseases Characteristics Clinical Complex Dementia Diagnosis Disease Disease Progression Drug Targeting Elderly Endosomes Event Foundations Goals Healthcare Systems Hippocampus Hour Human Immunoprecipitation Individual Induced pluripotent stem cell derived neurons Label Late Onset Alzheimer Disease Mass Spectrum Analysis Measurement Measures Membrane Proteins Methionine Methods Molecular Molecular Conformation Morphology Neurodegenerative Disorders Neurofibrillary Tangles Neurons Older Population Organoids Paint Pathogenesis Pathologic Patients Peptides Persons Pharmaceutical Preparations Phosphorylation Physiologic pulse Play Protein Analysis Protein Biosynthesis Protein Subunits Proteins Quality of life Reaction Reporting Research Resolution Risk Factors Role Sampling Senile Plaques Solvents Structure Techniques Technology Testing abeta accumulation age related aging population amyloid precursor protein processing analog baby boomer brain tissue cohort combat drug candidate drug development experimental study extracellular human disease hyperphosphorylated tau in vivo induced pluripotent stem cell network dysfunction new therapeutic target novel prevent protein complex protein expression protein folding protein protein interaction protein structure reduce symptoms small molecule tau Proteins tau aggregation theories trafficking

项目摘要

Summary/Abstract Late Onset Alzheimer's disease (LOAD) is the most common form of age-related dementia. Currently, 5 million people in the US are afflicted with LOAD and with the aging population this number is expected to double in the next decades. There are no drugs to cure or halt the progression of LOAD. The two pathological hallmarks of LOAD are extracellular amyloid plaques formed by the insoluble A peptide and neurofibrillary tangles consisting of hyperphosphorylated Tau protein. To date, most clinical drug candidates have targeted A, but none have proven effective at ameliorating the symptoms of AD. New drug targets are needed to combat this increasingly common and devastating disease. The recent identification of LOAD risk factors has revealed an enrichment of proteins in the endosomal-lysosomal network (ELN). This corroborates decades of evidence that the disruption of the ELN is an early event in LOAD pathogenesis and indicates that the ELN contains potential drug targets. However, a lack of molecular characterization of ELN has prevented the discovery of suitable candidates. Once considered “undruggable”, protein-protein interactions (PPI) are emerging as attractive targets for drug development. Global analysis of PPI in the ELN has not been studied. We propose to use mass spectrometry to quantitate different biochemical characteristics ELN protein complexes and determine how LOAD induces alterations in PPI within the ELN. These experiments will be performed in human brain tissues (AD vs. age-matched controls) and in human AD and control IPSC derived neurons and organoids. Endogenous ELN targets will be immunoprecipitated with validated antibodies and quantitated between conditions to identify novel ELN interactors and disease-relevant drug targets. Quantitation will be performed using multiplexing isobaric labeling technology. The structure of the ELN complexes will also be resolved, and differences will be quantitated with using our Covalent Protein Painting method. Additionally, we will employ a novel application of the non-canonical amino acid, azidohomoalanine, to quantitate the stability of ELN protein complexes. Our proposal will produce three different quantitative measurements of the influence of LOAD pathogenesis on ELN protein complexes and will provide alternative drug development targets for the most common form of age-related dementia.

摘要/摘要晚发性阿尔茨海默病(LOAD)是年龄相关性痴呆最常见的形式。目前，美国有500万人饱受负担和人口老龄化的困扰预计在未来几十年，这一数字将翻一番。没有药物可以治愈或阻止这种疾病负荷的递增。LOAD的两个病理特征是细胞外淀粉样斑块由不溶性A肽和过度磷酸化组成的神经原纤维缠结形成 Tau蛋白。到目前为止，大多数临床候选药物都针对A，但没有人被证明能有效改善AD的症状。需要新的药物靶点来对抗这一点越来越常见和破坏性的疾病。最近对负荷风险因素的识别已经揭示了蛋白质在内体-溶酶体网络(ELN)中的丰富。这证实了数十年来的证据，即ELN的中断是负载的早期事件并表明ELN含有潜在的药物靶点。然而，缺乏 ELN的分子特征阻止了合适的候选者的发现。一次被认为是“无法下药”的蛋白质-蛋白质相互作用(PPI)正在成为吸引人的靶点药物开发。对ELN中购买力平价的全球分析尚未得到研究。我们建议使用质谱法对不同生化特征的ELN蛋白复合体和确定负载如何引起ELN内PPI的变化。这些实验将是在人脑组织(AD与年龄匹配的对照组)以及人类AD和对照组中进行 IPSC来源于神经元和有机体。内源性ELN靶点将与验证抗体并在条件之间进行定量以确定新的ELN相互作用元件和与疾病相关的药物靶点。将使用多路传输等压标记来执行定量技术ELN复合体的结构也将得到解决，差异将被解决使用我们的共价蛋白质涂抹方法进行定量。此外，我们还将利用一部小说应用非正则氨基酸叠氮高丙氨酸定量测定ELN的稳定性蛋白质复合体。我们的建议将产生三种不同的量化测量 LOAD发病机制对ELN蛋白复合体的影响及可能提供的替代药物针对最常见形式的年龄相关性痴呆的发展目标。