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

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.

项目概要阿尔茨海默氏病 (AD) 是一种进行性神经退行性疾病，是导致以下疾病的最常见原因在美国，载脂蛋白 E 基因 (APOE) 的变异与痴呆症密切相关。在 AD 的发展中，APOE 主要在星形胶质细胞中合成。将脂质和胆固醇从星形胶质细胞输送到其他细胞类型，大多数人都具有主要等位基因之一。 APOE、APOE ε2 (R176C)、APOE ε3（参考等位基因）和 APOE ε4 (C130R) 这些错义变体是。与不同程度的阿尔茨海默病风险相关，其中 APOE ε4 增加疾病风险，而 APOE ε2 降低 AD 风险最近一项对 100,000 多名患者进行基因分型的分析表明，这种情况很少见。错义变异与不同程度的阿尔茨海默病风险相关。 APOE 的变异如何改变星形胶质细胞的生理机能从而导致阿尔茨海默病尚不清楚。试图理解这一点，我们之前的工作采用基于质谱的热蛋白质组分析来我们使用酵母作为简化模型来确定错义变异导致的蛋白质相互作用的变化。能够显示突变热蛋白质组分析，以解决亚复合物水平的蛋白质-蛋白质相互作用由于错义变异导致的蛋白质复合物的破坏研究这些罕见的错义变异的作用。关于星形胶质细胞中的 APOE，我们将采用小鼠原代星形胶质细胞培养物，该培养物将产生人类 APOE 首先，我们将确定罕见的错义变异对脂蛋白的影响。我们将确定 APOE 变体摄取的差异。 Aβ 肽、脂化状态以及对星形胶质细胞转录组的影响。使用突变热蛋白质组分析来检测 APOE 变体导致的蛋白质相互作用的变化总而言之，这些数据将扩展我们对错义变异如何变化的理解。改变 APOE 的功能并有助于深入了解导致阿尔茨海默病的分子机制疾病。