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：
8877782
负责人：
Amal F Khalil Kaddoumi
金额：
$ 29.2万
依托单位：
UNIVERSITY OF LOUISIANA AT MONROE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8877782
关键词：
Alzheimer&apos s Disease Amyloid Amyloid beta-Protein Blood - brain barrier anatomy Blood Vessels Brain Cell Culture Techniques Cell Line Cell physiology Cerebral Amyloid Angiopathy Clinical Clinical Research Data Data Analyses Dementia Deposition Development Disease Drainage procedure Educational process of instructing Elderly Endothelial Cells Frequencies Functional disorder Funding Future Goals Gold Health Health Care Costs Homeostasis In Vitro Investigation Kinetics Laboratories Lead Libraries Mediating Mentors Methodology Methods Modeling Molecular Nerve Degeneration Neuronal Dysfunction Outcome Pathogenesis Pathway interactions Permeability Pharmaceutical Preparations Pharmacy Schools Prevention Public Health Reporting Reproducibility Role Secure Students Techniques Testing Therapeutic Tight Junctions Training Validation Vascular Dementia Vascular Diseases Wild Type Mouse Work abeta accumulation base cerebrovascular drug development drug discovery high throughput screening improved in vivo lucifer yellow mouse model novel novel therapeutics prevent protein transport public health relevance screening small molecule small molecule libraries standard measure statistics therapeutic target therapy development

项目摘要

DESCRIPTION (provided by applicant): Targeting cerebrovascular endothelial cells as a therapeutic approach for amyloid pathogenesis. The accumulation of amyloid-β (Aß) 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 Aß in dementias such as AD. While the basis by which Aß mediates deleterious effect on the blood-brain barrier (BBB) is likely multifactorial, numerous studies indicate a role for Aß mediated increases in endothelial cell permeability. The exact causes for BBB dysfunction in CAA are not well known, however impaired clearance of Aß from the brain across the BBB as well as a reduction in the efficacy of the perivascular drainage of Aβ 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 Aß, the availability of drugs to treat Aß 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 Aß 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 Aß 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 Aβ clearance and reduce Aβ 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 Aβ 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 Aβ42 oligomers, for their effect on the gross permeability of cerebrovascular endothelial cells using Lucifer Yellow (LY) as a gross permeability marker. Compounds which reduce Aβ-mediated permeability will be advanced to Sub-Aim 1B; 1B) identify stimulators of Aβ clearance across the BBB model using the gold standard measure of iodinated Aβ as the endpoint for Aβ 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 Aβ 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 Aβ42 oligomers induced permeability and reduced Aβ clearance in primary cerebrovascular endothelial cells with those of Sub-Aim 2A, 2C) mechanistic investigation for improved BBB tightness, integrity and Aβ 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 Aβ accumulation, and BBB integrity in a mouse model of CAA. Methods and techniques to be used to accomplish the above aims include in vitro cell culture, high-throughput screening, transport, permeability and clearance studies, Aβ 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.

描述（由适用提供）：靶向脑血管内皮细胞作为淀粉样发病机理的治疗方法。淀粉样蛋白β（Aß）在脑血管中的积累会导致脑淀粉样血管病（CAA）的发展。 CAA是高频与阿尔茨海默氏病（AD）同时存在的病理特征，强调了血管Aß在痴呆症（例如AD）中的潜在重要作用。尽管Aß介导有害作用对血脑屏障（BBB）的基础可能是多因素的，但大量研究表明Aß介导的内皮细胞渗透性的增加起作用。 BBB功能障碍在CAA中的确切原因尚不清楚，但是已经提出，Aβ的大脑清除Aß从大脑中清除Aß的清除率降低了Aβ的多发性Aβ的有效性，以增强旧旧的脑血管和障碍性淀粉样淀粉样板的积累。尽管我们了解了导致BBB功能障碍和Aß清除率的途径，但仍缺乏治疗Aß发病机理的药物的可用性，CAA和AD仍然缺乏。该项目的长期目标是开发以BBB为目标的理论，以恢复其功能并最大程度地从大脑中清除Aß，这对于预防或延迟CAA和AD的发作很重要。该项目是将实验性内皮BBB模型完全表征为有效的高通量筛查（HTS）格式，以鉴定有关血管Aß发病机理（AD，CAA和血管痴呆）的治疗。中心假设是使用高度新颖的脑血管内皮内皮BBB模型的高通量筛选（HTS）可用于鉴定有益地调节Aβ清除率并减少Aβ介导的BBB通透性增加的小分子。我们将通过追求以下特定目的来检验这一假设：1）利用基于细胞系的BBB模型来筛选内皮细胞功能Aβ介导的灾害的调节剂。通过研究Aβ42低聚物的情况，将实现此目标：1a）筛选化合物的化合物，因为它们对使用Lucifer黄色（LY）作为总渗透性标记的脑血管内皮细胞的总渗透性的影响。降低Aβ介导的渗透性的化合物将推进Sub-Aim 1b； 1b）使用碘化Aβ的金标准测量作为Aβ清除率的终点，识别BBB模型跨BBB模型的Aβ清除率的刺激剂。 AIM 2）将进一步研究命中化合物。和Aβ清除蛋白（运输和降解）。该目标将通过测试以下子iMS：2a）进行次要确认并确定目标1、2b中确定的命中的概况，比较命中动力学以改善Aβ42低聚剂诱导的渗透率，诱导的渗透率可诱导的aβ渗透率，并降低了脑脑内膜内皮症的脑脑内部和bb bbiaim 2A，2A，2A，2A，2CA，2CA，2C）的Aβ清除率通过从2B选择的HIT化合物的Aβ清除率。 3）从AIM 2到体内调节血管和副群Aβ积累的测试能力，以及CAA小鼠模型中的BBB完整性。用于完成上述目的的方法和技术包括体外细胞培养，高通量筛查，运输，渗透性和清除研究，Aβ动力学，从野生型小鼠的大脑中分离的微丝以及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。