Targeting cerebrovascular endothelial cells as a therapeutic approach for amyloid pathogenesis

靶向脑血管内皮细胞作为淀粉样蛋白发病机制的治疗方法

基本信息

批准号：
9548880
负责人：
Amal F Khalil Kaddoumi
金额：
$ 12.77万
依托单位：
AUBURN UNIVERSITY AT AUBURN
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9548880
关键词：
Targeting cerebrovascular endothelial cells therapeutic

项目摘要

Targeting cerebrovascular endothelial cells as a therapeutic approach for amyloid pathogenesis Project Summary The accumulation of amyloid- (Aβ) in the brain blood vessels can result in the development of cerebral amyloid angiopathy (CAA). CAA is a pathological feature present concomitantly with Alzheimer’s disease (AD) at a high frequency, highlighting a potentially important role for vascular Aβ in dementias such as AD. While the basis by which Aβ mediates deleterious effect on the blood-brain barrier (BBB) is likely multifactorial, numerous studies indicate a role for Aβ mediated increases in endothelial cell permeability. The exact causes for BBB dysfunction in CAA are not well known, however impaired clearance of Aβ from the brain across the BBB as well as a reduction in the efficacy of the perivascular drainage of A have been proposed to enhance accumulation of cerebrovascular and parenchymal amyloid deposits in the elderly. Despite our understanding of the pathways responsible for BBB dysfunction and clearance of Aβ, the availability of drugs to treat Aβ pathogenesis related disorders, CAA and AD, remains lacking. The long-term goal of this project is to develop therapeutics that target the BBB to restore its function and maximize clearance of Aβ from the brain, which is important to prevent or delay onset of CAA and AD. The overall goal of this project is to fully characterize an experimental endothelial BBB model as an effective high-throughput screening (HTS) format to identify therapeutics for vascular Aβ pathogenesis disorders (AD, CAA and vascular dementia). The central hypothesis is that high-throughput screening (HTS) utilizing a highly novel cerebrovascular endothelial BBB model can be used to identify small molecules which beneficially regulate A clearance and reduce A mediated increases in BBB permeability. We will test this hypothesis by pursuing the following specific aims: 1) Utilization of a cell line-based BBB model to screen for modulators of A mediated disturbances of endothelial cell function. This aim will be accomplished by investigating the following sub-aims: 1A) screen for compounds, in the presence of A42 oligomers, for their effect on the gross permeability of cerebrovascular endothelial cells using Lucifer Yellow (LY) as a gross permeability marker. Compounds which reduce A-mediated permeability will be advanced to Sub-Aim 1B; 1B) identify stimulators of A clearance across the BBB model using the gold standard measure of iodinated A as the endpoint for A clearance. Hit compounds will be further examined in Aim 2. 2) Validation and mechanistic investigation of hit compounds from Aim1 for their ability to modulate expression of tight junction and A clearance proteins (transport and degradation). This aim will be accomplished by testing the following sub-aims: 2A) conduct secondary confirmation and establish profiles for hits identified in Aim 1, 2B) comparison of hits kinetics for amelioration of A42 oligomers induced permeability and reduced A clearance in primary cerebrovascular endothelial cells with those of Sub-Aim 2A, 2C) mechanistic investigation for improved BBB tightness, integrity and A clearance by hit compounds selected from 2B. 3) Test ability of the top 2 hits from Aim 2 to in vivo modulate vascular and parenchymal A accumulation, and BBB integrity in a mouse model of CAA. Methods and technique to be used to accomplish the above aims include in vitro cell culture, high-throughput screening, transport, permeability and clearance studies, A kinetics, microvessels isolation from brains of wild type mice, and in vivo studies in CAA model. The data produced will provide candidate therapeutic molecules to test in future clinical studies.

靶向脑血管内皮细胞作为淀粉样发病机理的治疗方法项目摘要淀粉样蛋白 - （Aβ）在脑血管中的积累可能导致大脑的发展淀粉样血管病（CAA）。 CAA是与阿尔茨海默氏病（AD）同时存在的病理特征在高频率下，突出了血管Aβ在痴呆症（例如AD）中的潜在重要作用。尽管 Aβ介导有害影响对血脑屏障（BBB）的基础可能是多因素的大量研究表明，Aβ介导的内皮细胞渗透性的增加起作用。确切的原因对于CAA中的BBB功能障碍，尚不清楚，但是从大脑中清除Aβ的障碍已经提出了BBB以及A的血管周围排水效率的降低以增强古老的脑血管和副淀粉样蛋白沉积物的积累。尽管我们的理解负责BBB功能障碍和Aβ清除率的途径，治疗Aβ的药物的可用性与发病机理相关的疾病，CAA和AD仍然缺乏。该项目的长期目标是开发靶向BBB以恢复其功能并最大程度地清除Aβ的治疗对于预防或延迟CAA和AD的发作很重要。该项目的总体目标是完全表征实验性内皮BBB模型作为有效的高通量筛选（HTS）格式以识别血管Aβ发病机理（AD，CAA和血管痴呆）的治疗剂。中心假设是使用高度新颖的脑血管内皮BBB模型的高通量筛查（HTS）可以是用于鉴定有益地调节清除率并减少介导的增加的小分子 BBB渗透性。我们将通过追求以下特定目的来检验这一假设：1）使用单元基于线的BBB模型，以筛选内皮细胞功能的A介导的灾难的调节器。这 AIM将通过调查以下子AIMS来实现：1a）在存在下的化合物屏幕 A42低聚物的影响，因为它们对使用Lucifer的脑血管内皮细胞的总渗透性的影响黄色（LY）作为总渗透性标记。降低A介导的渗透性的化合物将是先进到Sub-aim 1b； 1b）使用黄金识别BBB模型中A清除率的刺激器碘化A的标准测量作为清除的终点。命中化合物将进一步研究目标2。2）AIM1的验证和机械投资的调节能力紧密连接和A清除蛋白的表达（运输和降解）。这个目标将是通过测试以下子AIMS来完成：2a）进行次要确认并建立个人资料 AIM 1，2B中鉴定出的命中率）比较击中动力学的A42低聚物诱导的渗透性并降低了用Sub-Aim 2a，2C的原代脑血管内皮细胞中的A清除率）通过选定的命中化合物提高BBB紧密度，完整性和A清除的机械研究从2B。 3）从AIM 2到体内调节血管和副群的测试能力 CAA小鼠模型中的积累和BBB完整性。用于完成的方法和技术以上目的包括体外细胞培养，高通量筛查，运输，渗透性和清除率研究，A动力学，从野生型小鼠的大脑中分离的微血管以及CAA模型中的体内研究。所产生的数据将为候选治疗分子提供在未来的临床研究中测试。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Oleocanthal-rich extra-virgin olive oil enhances donepezil effect by reducing amyloid-β load and related toxicity in a mouse model of Alzheimer's disease.

DOI：
10.1016/j.jnutbio.2017.12.006
发表时间：
2018-05
期刊：
The Journal of nutritional biochemistry
影响因子：
0
作者：
Batarseh YS;Kaddoumi A
通讯作者：
Kaddoumi A

Plasma Rich in Growth Factors (PRGF) Disrupt the Blood-Brain Barrier Integrity and Elevate Amyloid Pathology in the Brains of 5XFAD Mice.

富含生长因子 (PRGF) 的血浆会破坏 5XFAD 小鼠大脑中的血脑屏障完整性并增强淀粉样蛋白病理。

DOI：
10.3390/ijms20061489
发表时间：
2019
期刊：
International journal of molecular sciences
影响因子：
5.6
作者：
Duong,Quoc-Viet;Kintzing,MargiaL;Kintzing,WilliamE;Abdallah,IhabM;Brannen,AndrewD;Kaddoumi,Amal
通讯作者：
Kaddoumi,Amal

Crocus sativus Extract Tightens the Blood-Brain Barrier, Reduces Amyloid β Load and Related Toxicity in 5XFAD Mice.

DOI：
10.1021/acschemneuro.7b00101
发表时间：
2017-08-16
期刊：
ACS chemical neuroscience
影响因子：
5
作者：
Batarseh YS;Bharate SS;Kumar V;Kumar A;Vishwakarma RA;Bharate SB;Kaddoumi A
通讯作者：
Kaddoumi A

Author Correction: Amylin and pramlintide modulate γ-secretase level and APP processing in lipid rafts.

作者更正：胰淀素和普兰林肽调节脂筏中的γ分泌酶水平和APP加工。

DOI：
10.1038/s41598-020-68281-y
发表时间：
2020
期刊：
Scientific reports
影响因子：
4.6
作者：
Mousa,YoussefM;Abdallah,IhabM;Hwang,Misako;Martin,DouglasR;Kaddoumi,Amal
通讯作者：
Kaddoumi,Amal

Characterization of Hit Compounds Identified from High-throughput Screening for their Effect on Blood-brain Barrier Integrity and Amyloid-β Clearance: In Vitro and In Vivo Studies.

DOI：
10.1016/j.neuroscience.2018.03.028
发表时间：
2018-05-21
期刊：
Neuroscience
影响因子：
3.3
作者：
Elfakhri KH;Duong QV;Langley C;Depaula A;Mousa YM;Lebeouf T;Cain C;Kaddoumi A
通讯作者：
Kaddoumi A