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) 是最常见的与年龄相关的痴呆症。目前，美国有 500 万人受到 LOAD 的影响，并且随着人口老龄化，预计在未来几十年内这一数字将增加一倍，目前还没有药物可以治愈或阻止这种疾病。 LOAD 的两个病理特征是细胞外淀粉样斑块。由不溶性 A 肽和由过度磷酸化组成的神经原纤维缠结形成迄今为止，大多数临床候选药物都以 A 为靶点，但尚未得到证实。需要新的药物靶标来有效改善 AD 症状。最近发现的 LOAD 危险因素已变得越来越常见和具有破坏性。揭示了内体-溶酶体网络（ELN）中蛋白质的富集。证实了数十年的证据，表明 ELN 的破坏是 LOAD 的早期事件发病机制并表明 ELN 含有潜在的药物靶点。 ELN 的分子表征阻碍了合适候选物的发现。被认为“不可成药”的蛋白质-蛋白质相互作用（PPI）正在成为有吸引力的目标尚未研究 ELN 中 PPI 的药物开发。质谱法来量化不同生化特征的 ELN 蛋白复合物和确定 LOAD 如何引起 ELN 内 PPI 的变化。在人类脑组织（AD 与年龄匹配的对照）以及人类 AD 和对照中进行 IPSC 衍生的神经元和类器官将用免疫沉淀法进行免疫沉淀。验证抗体并在不同条件下进行定量，以识别新的 ELN 相互作用因子将使用多重同量异位标记进行定量。 ELN复合体的结构也将得到解决，差异也将得到解决。此外，我们将采用一种新颖的共价蛋白质绘画方法进行定量。应用非规范氨基酸叠氮高丙氨酸来定量 ELN 的稳定性我们的建议将产生三种不同的定量测量。 LOAD发病机制对ELN蛋白复合物的影响并将提供替代药物最常见形式的年龄相关性痴呆的发展目标。