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）的每个细胞成分功能障碍 – 血管细胞、神经胶质细胞和神经元 – 在实验中已被证实与阿尔茨海默病 (AD) 的进化有关，这些发现导致了“神经血管”的出现。 AD 假说认为，脑血管功能障碍会导致认知能力下降和痴呆 AD 项目 3 支持 P01 的总体主题，重点关注载脂蛋白 E (APOE) 基因，这是主要的遗传基因。 AD 的危险因素，通过在细胞、分子水平上通过实验询问 BBB/神经血管假说和系统水平，并使用新型 APOE3 和 APOE4knock 以异构体特异性和性别特异性方式在 (KI)flox/flox 小鼠 (E3F; E4F) 中，有或没有从星形胶质细胞和周细胞（apoE 的关键来源）缺失 BBB 相关的 apoE），以及有或没有 Aβ 和 tau 病理学基于我们的试验和已发表的数据。 1）血脑屏障（BBB）蛋白质相互作用（PPI）信号网络被破坏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在新的 APOE KIF 系中，相对于 APOE3，APOE4 驱动的功能障碍和行为变化均没有以及 AD 病理学；2) 3K3A 激活蛋白 C (APC)（APC 的细胞信号传导类似物）将纠正 BBB 和 PSD95 突触 PPI 信号网络失调，将保护 BBB 和神经元功能，并且减少 AD 病理，从而延迟 APOE/AD 模型的发病并减缓疾病进展。检验我们的假设，我们将单独使用新型 E3F 和 E4F 小鼠，并与 i) 与特定的 Cre 品系杂交在星形胶质细胞和周细胞中表达 Cre 重组酶，以确定细胞特异性删除密钥的效果 BBB 相关的 apoE 来源和 ii) 与 APP/PS1-21 小鼠和 P301S tau 小鼠一起研究相互作用为了评估 BBB/血管功能障碍，我们将使用：1) 先进的分子技术。通过同时测量 CSF 中的 BBB/NVU 细胞特异性以及 Aβ 和 tau 生物标志物来评估 BBB 和血浆；2) 新型 BBB 蛋白质组学分析；3) BBB 和 CBF 的高级 MRI 评估；通过组织分析验证生物流体生物标志物，我们将评估突触和神经元功能障碍。使用：5) PSD95 相互作用组分析；6) Papez 环路/DMN 中海马通路的病毒道追踪； 6) 行为和 7) 神经病理学分析我们将评估 apoE 等位基因对 BBB 和的特异性影响。生物流体、组织和成像生物标志物、BBB 和 PSD95 PPI 分析、海马的神经元功能障碍不同年龄的有和没有 apoE 缺失的 APOE KIF 小鼠（雄性、雌性）的病毒道追踪和行为来自星形胶质细胞和周细胞 (AIM 1)，与 APP/PS1-21 和 P301S tau 小鼠杂交，与 AD 病理学相关 (AIM 2)，并将评估不同年龄的 APOE KIF 小鼠（雄性、雌性）的 3K3A-APC 治疗，单独治疗和与 APP/PS1-21 和 P301S tau 小鼠 (AIM 3) 杂交将加深我们对 tau 蛋白的理解。 AD 的发病机制在细胞和分子水平上以 apoE 亚型特异性和性别特异性方式进行。