Proteomic and functional analysis of missense variants of APOE associated with Alzheimer disease risk

与阿尔茨海默病风险相关的 APOE 错义变异的蛋白质组学和功能分析

基本信息

批准号：
10536747
负责人：
HR Sagara Wijeratne
金额：
$ 3.74万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
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：
2022-09-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10536747
关键词：
Abeta clearance Affect Affinity Chromatography Age Alleles Alzheimer&apos s Disease Alzheimer&apos s disease risk Amino Acid Sequence Amyloid beta-Protein Apolipoprotein E Apolipoproteins Astrocytes Binding Biochemical Biological Biology Biophysics Brain Cholesterol Cholesterol Homeostasis Clinical Data Dementia Dependovirus Development Diagnostic Doctor of Philosophy E protein Environment Genes Genotype Human Individual Knockout Mice Language Lead Lipids Lipoproteins Mass Spectrum Analysis Memory Mentors Metabolism Modeling Molecular Mus Mutation Neocortex Neurodegenerative Disorders Pathogenesis Pathway interactions Patients Peptides Phenotype Physicians Physiological Physiology Plasmids Post-Translational Protein Processing Property Protein Isoforms Proteins Proteome Proteomics Regulation Reporting Research Resources Risk Role Scientist Senile Plaques Serotyping Synaptic plasticity System Techniques Temperature Therapeutic Time United States Variant Work Yeasts abeta accumulation apolipoprotein E-3 base biophysical properties cell type disorder risk executive function extracellular genetic risk factor genetic variant high risk innovation insight lipid transport melting mild cognitive impairment mutant neurofibrillary tangle formation neurotoxicity prognostic protein complex protein folding protein protein interaction rare variant receptor receptor binding targeted treatment transcriptome transcriptome sequencing transcriptomics uptake

Abeta clearance Affect Affinity Chromatography Age Alleles Alzheimer&apos s Disease Alzheimer&apos s disease risk Amino Acid Sequence Amyloid beta-Protein Apolipoprotein E Apolipoproteins Astrocytes Binding Biochemical Biological Biology Biophysics Brain Cholesterol Cholesterol Homeostasis Clinical Data Dementia Dependovirus Development Diagnostic Doctor of Philosophy E protein Environment Genes Genotype Human Individual Knockout Mice Language Lead Lipids Lipoproteins Mass Spectrum Analysis Memory Mentors Metabolism Modeling Molecular Mus Mutation Neocortex Neurodegenerative Disorders Pathogenesis Pathway interactions Patients Peptides Phenotype Physicians Physiological Physiology Plasmids Post-Translational Protein Processing Property Protein Isoforms Proteins Proteome Proteomics Regulation Reporting Research Resources Risk Role Scientist Senile Plaques Serotyping Synaptic plasticity System Techniques Temperature Therapeutic Time United States Variant Work Yeasts abeta accumulation apolipoprotein E-3 base biophysical properties cell type disorder risk executive function extracellular genetic risk factor genetic variant high risk innovation insight lipid transport melting mild cognitive impairment mutant neurofibrillary tangle formation neurotoxicity prognostic protein complex protein folding protein protein interaction rare variant receptor receptor binding targeted treatment transcriptome transcriptome sequencing transcriptomics uptake

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项目摘要

PROJECT SUMMARY Alzheimer’s disease (AD) is a progressive neurodegenerative disease that is the most common cause of dementia in the United States. Variants in the apolipoprotein E gene (APOE) is strongly associated with development of AD. In the brain, APOE is mostly synthesized in the astrocytes. The primary role of APOE is to deliver lipids and cholesterol from astrocytes to other cell types. Most humans have one of the major allele of APOE, APOE ε2 (R176C), APOE ε3 (reference allele), and APOE ε4 (C130R). These missense variants are associated with different levels of Alzheimer disease risks where APOE ε4 increases the risk of disease whereas APOE ε2 decrease risk of AD. A recent analysis that genotyped over 100,000 patients showed that rare missense variants were associated with different levels of Alzheimer disease risk. The molecular mechanism of how variants in APOE changes astrocyte physiology that could lead to Alzheimer disease is not well known. To attempt to understand this, our previous work adapted mass spectrometry-based thermal proteome profiling to determine changes in protein interaction as a result of missense variants. Using yeast as simplified model, we were able to show mutant thermal proteome profiling to resolve subcomplex level protein-protein interaction disruptions of a protein complexes due to missense variants. To study the role of these rare missense variants on APOE in astrocytes, we will employ an mouse primary astrocyte culture that will generate human APOE with the specific missense variant. First, we will determine the effect of the rare missense variants on the lipoprotein functions and astrocyte physiology. We will determine the differences between the APOE variants of uptake of Aβ peptides, lipidation status, and the effects on the astrocyte transcriptome. Second, we will determine the changes in protein interactions as a result in APOE variants using mutant thermal proteome profiling to detect thermal stability changes. Taken together, these data will expand our understanding of how missense variants change the functions of APOE and help give insight into the molecular mechanisms that result in Alzheimer disease.

项目摘要阿尔茨海默氏病（AD）是一种进行性神经退行性疾病，是最常见的原因美国的痴呆症。载脂蛋白E基因（APOE）中的变体与广告的开发。在大脑中，APOE主要是在星形胶质细胞中合成的。 Apoe的主要作用是将脂质和胆固醇从星形胶质细胞传递到其他细胞类型。大多数人都有一个主要等位基因之一 APOE，APOEε2（R176C），APOEε3（参考等位基因）和APOEε4（C130R）。这些错义变体是与不同水平的阿尔茨海默氏病风险相关 APOEε2降低了AD的风险。最近的分析表明，超过100,000名患者表明罕见错义变体与不同水平的阿尔茨海默氏病风险有关。分子机制 APOE的变体如何改变可能导致阿尔茨海默氏病的星形胶质细胞生理学。到试图理解这一点，我们以前的工作调整了基于质谱的热蛋白质组分析确定由于错义变异的结果，蛋白质相互作用的变化。使用酵母作为简化模型，我们能够显示突变的热蛋白分析以解决亚复杂水平蛋白 - 蛋白质相互作用由于错义变体引起的蛋白质复合物的破坏。研究这些稀有错义变体的作用在星形胶质细胞中的Apoe上，我们将采用小鼠原代星形胶质细胞培养特定的错义变体。首先，我们将确定稀有错义变体对脂蛋白的影响功能和星形胶质细胞生理。我们将确定摄取的APOE变体之间的差异 Aβ肽，脂质状态以及对星形胶质细胞转录组的影响。第二，我们将确定蛋白质相互作用的变化是APOE变体使用突变的热蛋白质组分析检测的APOE变体热稳定性变化。综上所述，这些数据将扩展我们对错义变体的理解更改APOE的功能，并有助于深入了解导致阿尔茨海默氏症的分子机制疾病。