Project 3 - Animal Models Examining Neurovasculature

项目 3 - 检查神经脉管系统的动物模型

基本信息

批准号：
10621719
负责人：
Berislav V Zlokovic
金额：
$ 87.4万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-30 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621719
关键词：
APP-PS1 Age Alleles Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease risk Amyloid beta-Protein Animal Model Apolipoprotein E Astrocytes Behavior Behavioral Biological Markers Blood - brain barrier anatomy Blood Vessels Blood brain barrier dysfunction Cells Cerebrovascular Circulation Cerebrovascular Disorders Cognition DLG4 gene Data Dementia Development Disease Progression Endothelium Enterobacteria phage P1 Cre recombinase Evolution Female Functional disorder Funding Opportunities Gender Generations Genes Hippocampus Image Impaired cognition Instruction Ischemic Stroke Knock-in Link LoxP-flanked allele Magnetic Resonance Imaging Measures Mediating Molecular Mus Neuroglia Neuronal Dysfunction Neurons Pathogenesis Pathologic Pathology Pathway Analysis Pathway interactions Pericytes Phase Plasma Protein Isoforms Proteomics Publishing Signal Transduction Source Synapses System Testing Tissues Transgenic Model Validation Vascular Dementia Vascular Diseases Viral activated Protein C analog apolipoprotein E-3 apolipoprotein E-4 blood-brain barrier disruption cerebrovascular dementia risk epidemiology study genetic risk factor imaging biomarker male mouse model neuropathology neurovascular neurovascular unit novel protein protein interaction tau Proteins tissue biomarkers

项目摘要

PROJECT 3 – PROJECT SUMMARY/ ABSTRACT Dysfunctionin the blood-brain barrier (BBB) and each of the cellular components of the neurovascular unit (NVU) – vascular cells, glial cells, and neurons – has been linked to Alzheimer’s disease (AD) evolution in experimental, imaging, pathological, and epidemiological studies. These findings have led to an emerging ‘neurovascular hypothesis’ of AD which holds that cerebrovascular dysfunction contributes to cognitive decline and dementia in AD. Project 3 supports the overall theme of P01 focused on apolipoprotein E (APOE) gene, the major genetic risk factor for AD, by interrogating the BBB/neurovascular hypothesis experimentally at the cellular, molecular and systems levels and in an isoform-specific and gender-specific fashion using novel APOE3 and APOE4knock in (KI)flox/flox mice (E3F; E4F) with and without apoE deletion from astrocytes and pericytes (the key sources of BBB-associated apoE), and with and without Aβ and tau pathology. Based on our pilot and published data we hypothesize that 1) Disrupted blood-brain barrier (BBB) protein-protein interaction (PPI) signaling networks underlying endothelial barrier disruption and BBB dysfunction will precede and predict synaptic and neuronal dysfunction and behavioral changes driven by APOE4 relative to APOE3 in new APOE KIF lines both without and with AD pathology; and 2) 3K3A-activated protein C (APC), a cell signaling analog of APC, will correct dysregulated BBB and PSD95 synaptic PPI signaling networks, will protect BBB and neuronal function, and diminish AD pathology, thereby delaying onset and slowing down disease progression in APOE/AD models. To test our hypothesis, we will use novel E3F and E4F mice alone and crossed i) with Cre lines that specifically express Cre recombinase in astrocytes and pericytes to determine the effects of cell-specific deletion of the key BBB-associated apoE source(s) and ii) with APP/PS1-21 mice and P301S tau mice to investigate interactions with Aβ and tau pathways. To evaluate BBB/vascular dysfunction we will use: 1) advanced molecular assessment of the BBB by simultaneously measured BBB/NVU cell-specific and Aβ and tau biomarkers in CSF and plasma; 2) a novel BBB proteomics analysis; 3) advanced MRI assessment of BBB and CBF; and 4) validation of biofluid biomarkers by tissue analysis. To evaluate synaptic and neuronal dysfunction we will use: 5) PSD95 interactomeanalysis; and 6) viral tract-tracing of hippocampal pathways in the Papez circuit/DMN; and 6) behavior and 7) neuropathology analysis. We will evaluate apoE-allele-specific effects on BBB and neuronal dysfunction by biofluid, tissue and imaging biomarkers, BBB and PSD95 PPI analysis, hippocampal viral tract-tracing, and behavior in APOE KIF mice(male, female) at different ages with and without apoE deletion from astrocytes and pericytes (AIM 1), crossed with APP/PS1-21 and P301S tau micein relation to AD pathology (AIM 2), and will evaluate 3K3A-APC therapy in APOE KIF mice (male, female) at different ages, alone and crossed with APP/PS1-21 and P301S tau mice (AIM 3). Project 3 will advance our understanding of the pathogenesis of AD at the cellular and molecular level in an apoE isoform-specific and gender-specific fashion.

项目3 - 项目摘要/摘要血脑屏障（BBB）和神经血管单元（NVU）的每个细胞成分中的功能障碍 - 血管细胞，神经胶质细胞和神经元 - 与实验性的阿尔茨海默氏病（AD）相关，成像，病理和流行病学研究。这些发现导致了新兴的‘神经血管 AD的假设'认为脑血管功能障碍有助于认知能力下降和痴呆在广告中。项目3支持P01的总体主题，该主题侧重于载脂蛋白E（APOE）基因，这是主要的通用通用性 AD的危险因素，通过在细胞分子的实验中询问BBB/神经血管假设和系统级别以及使用小说APOE3和APOE4KNOCK的同工型特异性和性别特定的方式在（ki）flox/flox小鼠（E3F; e4f）中，有和没有星形胶质细胞和周细胞的APOE删除（主要来源与BBB相关的APOE），并且没有Aβ和TAU病理。根据我们的飞行员和发布的数据，我们假设1）破坏血脑屏障（BBB）蛋白质 - 蛋白质相互作用（PPI）信号网络潜在的内皮屏障破坏和BBB功能障碍将先于并预测突触和神经元 APOE4相对于APOE3在新的ApoE KIF线中驱动的功能障碍和行为变化都没有并具有广告病理学； 2）3K3A激活的蛋白C（APC），APC的细胞信号类似物，将纠正 BBB和PSD95合成PPI信号网络失调，将保护BBB和神经元功能，并且减少AD病理学，从而延迟APOE/AD模型中疾病进展的延迟。到检验我们的假设，我们将单独使用新颖的E3F和E4F小鼠，并与CRE线越过I）在星形胶质细胞和周细胞中的表达CRE重组酶，以确定钥匙的细胞特异性缺失的影响与APP/PS1-21小鼠和P301S TAU小鼠相关的BBB相关APOE源和II）研究相互作用具有Aβ和TAU途径。为了评估BBB/血管功能障碍，我们将使用：1）高级分子通过简单测量CSF中的BBB/NVU细胞特异性和Aβ和TAU生物标志物评估BBB 和血浆； 2）一种新型的BBB蛋白质组学分析； 3）对BBB和CBF的高级MRI评估；和4）通过组织分析验证生物流体生物标志物。为了评估突触和神经元功能障碍，我们将使用：5）PSD95相互作用分析； 6）帕佩兹电路/DMN中海马途径的病毒式追踪；和6）行为和7）神经病理学分析。我们将评估Apoe-Allele特定对BBB的影响生物流体，组织和成像生物标志物，BBB和PSD95 PPI分析的神经元功能障碍，海马病毒道追踪和APOE KIF小鼠（雄性，女性）的行为，有或没有APOE缺失来自星形胶质细胞和周细胞（AIM 1），与APP/PS1-21和P301S Tau小质量与AD病理学关系（AIM 2），并将评估不同年龄，单独的年龄和与App/PS1-21和P301S Tau小鼠越过（AIM 3）。项目3将促进我们对 ApoE同工型特异性和性别特异性的AD在细胞和分子水平上的AD发病机理。