ApoE isoform-specific protein interactions in brain aging and neurodegeneration

ApoE 亚型特异性蛋白质在大脑衰老和神经退行性疾病中的相互作用

基本信息

批准号：
7229782
负责人：
Kathleen S Montine
金额：
$ 18.95万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2009-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7229782
关键词：
AIDS Dementia Complex Adult Affinity Age Age of Onset Alleles Alzheimer&apos s Disease Alzheimer&apos s disease risk Amino Acids Amyloid Amyotrophic Lateral Sclerosis Antioxidants Apolipoprotein E Apolipoproteins Biological Models Biomedical Research Brain Brain Diseases Brain region Cerebral cortex Cerebrospinal Fluid Cerebrum Clinical Code Coupled Craniocerebral Trauma Data Degenerative Disorder Dementia Development Disease Dose Elderly Emerging Technologies Exploratory/Developmental Grant Fostering Genes Genotype Human Immunochemistry Immunohistochemistry Individual Inflammation Inherited Intervention Isotopes Isotopically-Coded Affinity Tagging Late Onset Alzheimer Disease Mammals Measures Metabolism Modeling Multiple Sclerosis Mus Nerve Degeneration Neuraxis Neurodegenerative Disorders Neurofibrillary Tangles Neurologic Numbers Outcome Parkinson Disease Patients Peptides Physiological Positioning Attribute Post-Translational Regulation Predisposition Prevalence Process Protein Isoforms Proteins Proteomics Race Relative (related person)Research Personnel Risk Risk Factors Sampling Source Synaptosomes System Technology Testing Tissue Sample United States National Institutes of Health Validation Variant Western Blotting aging brain apolipoprotein E-3 apolipoprotein E-4 brain tissue cerebrovascular early onset follow-up functional genomics genetic association human data interest lipid metabolism liquid chromatography mass spectrometry new technology novel programs promoter protective effect protein protein interaction receptor response to injury synaptogenesis tool

项目摘要

DESCRIPTION (provided by applicant): A risk factor for several neurodegenerative diseases is inheritance of the epsilon 4 allele of apolipoprotein E (APOE). The best established increased risk with inheritance of epsilon 4 allele of APOE is for Alzheimer's disease (AD) where inheritance of the epsilon 2 allele also appears to be protective. Although the subject of much interest and study, the mechanism behind these differing risks remains to be fully explained. We hypothesize that one mechanism for the difference in risk of AD is due to isoform-specific differences in apoE-associated proteins. The recently developed approach of high-throughput quantitative proteomic analysis combining isotope-coded affinity tagging (ICAT) with peptide analysis by LC/MS-MS allows us to identify and compare the relative abundance of apoE-associated proteins from different sources. In this study, I will affinity purify apoE-associated proteins from 5 human brain regions and compare the composition and relative abundance of each apoE-associated protein among different APOE genotypes in elderly individuals without dementia, and between AD patients and control individuals of the same genotype. In addition, I will perform identical analyses for cerebral homogenates and synaptosomes prepared from young and old apoE targeted replacement mice expressing human apoE isoforms under the mouse apoE promoter. In each case I will (i) identify proteins by proteomic analysis of pooled tissue samples, (ii) confirm the presence of selected proteins by Western blot of the same pooled samples, and (iii) statistically validate these differences by Western blotting of individual samples, as well as further validation by immunohistochemistry. I have contributed to recent studies that have successfully employed this three-step approach to identify proteins that are elevated in lumbar cerebrospinal fluid from probable AD patients and proteins that are present in neurofibrillary tangles, and now propose to use this novel technology to investigate apoE isoforms. Completion of this study of apoE-associated proteins will identify proteins that may be contributing to apoE isoform-specific risk or protection and provide candidate proteins to follow up with more detailed mechanistic studies in models of neurodegenerative diseases.

描述（由申请人提供）：几种神经退行性疾病的危险因素是epsilon 4载脂蛋白E（APOE）的遗传。 Epsilon 4 ApoE等位基因的遗传最佳的风险最高的是阿尔茨海默氏病（AD），那里的Epsilon 2等位基因的遗传似乎也具有保护性。尽管引起了很多兴趣和研究的主题，但这些不同风险背后的机制仍有待解释。我们假设AD风险差异的一种机制是由于APOE相关蛋白的同工型特异性差异所致。最近开发的高通量定量蛋白质组学分析将同位素编码的亲和力标记（ICAT）与LC/MS-MS结合肽分析的方法使我们能够识别和比较来自不同来源的APOE相关蛋白的相对丰度。在这项研究中，我将从5个人脑区域纯化纯化与APOE相关的蛋白质，并比较没有痴呆症的老年人的不同APOE基因型之间每种APOE相关蛋白的组成和相对丰度，并且在AD患者之间以及在同一基因型的个体之间进行控制。此外，我还将对由小鼠APOE同工型在小鼠APOE启动子下表达人apoE同工型的年轻和老式apoE靶向替代小鼠制备的脑匀浆和突触体进行相同的分析。在每种情况下，我将（i）通过对合并组织样品的蛋白质组学分析鉴定蛋白质，（ii）通过蛋白质印迹通过同一合并样品的蛋白质确认选定的蛋白质，（iii）通过蛋白质斑点通过单个样品的蛋白质斑点来证实这些差异，以及通过免疫组织的进一步验证。我为最近的研究做出了贡献，这些研究已成功地采用了这种三步方法来鉴定来自可能的AD患者和神经原纤维缠结中存在的蛋白质中脑脑脊髓液升高的蛋白质，现在建议使用这种新型技术来研究APOE同种型。这项与APOE相关蛋白的研究的完成将确定可能有助于APOE同工型特异性风险或保护的蛋白质，并提供候选蛋白质，以跟进神经退行性疾病模型中更详细的机械研究。