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/摘要阿尔茨海默氏病晚期（负载）是与年龄相关的痴呆症最常见的形式。目前，美国有500万人患有负载，随着人口老龄化的折磨预计在未来几十年中的数字将翻一番。没有药物可以治愈或停止负载的进展。负载的两个病理标志是细胞外淀粉样斑块由不溶性的辣椒和神经原纤维缠结形成，由高磷酸化组成 tau蛋白。迄今为止，大多数临床药物候选者都针对A，但没有证明有效缓解AD的症状。需要新的药物目标来应对这一目标越来越常见和毁灭性的疾病。负载风险因素最近的识别具有揭示了内体溶菌体网络（ELN）中蛋白质的富集。这证实了数十年的证据表明ELN的破坏是负载中的早期事件发病机理，表明ELN包含潜在的药物靶标。但是，缺乏 ELN的分子表征阻止了合适的候选者的发现。一次蛋白质 - 蛋白质相互作用（PPI）被认为是“不可能的” 药物开发。 ELN中PPI的全球分析尚未研究。我们建议使用质谱法以量化不同的生化特征ELN蛋白复合物和确定负载如何诱导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和对照中进行 IPSC衍生的神经元和器官。内源性ELN目标将通过经过验证的抗体并在条件之间进行了定量，以识别新型ELN相互作用者和与疾病有关的药物靶标。定量将使用多路复用等速标记进行技术。 ELN复合物的结构也将得到解决，差异将是通过使用我们的共价蛋白绘画方法来定量。此外，我们将采用小说非典型氨基酸Azidohomoalanine的应用来定量ELN的稳定性蛋白质复合物。我们的建议将产生三个不同的定量测量负载发病机理对ELN蛋白复合物的影响，并将提供替代药物最常见形式与年龄相关痴呆的形式的发展目标。