Identifying targeting antibodies in an in vitro model of the human blood-brain barrier during ischemic stroke

在缺血性中风期间人血脑屏障的体外模型中识别靶向抗体

基本信息

批准号：
10796765
负责人：
Moriah Katt
金额：
$ 15.64万
依托单位：
WEST VIRGINIA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-08 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10796765
关键词：
3-Dimensional ABCB1 gene Acclimatization Acute Address Alteplase Analysis of Variance Animal Model Animals Antibodies Apical Apoptosis Architecture Area Astrocytes Autopsy Basal lamina Benchmarking Binding Biodistribution Biological Assay Blood Blood - brain barrier anatomy Blood brain barrier dysfunction Brain Brain Ischemia Brain region Brain-Derived Neurotrophic Factor CASP3 gene CRISPR interference Cadaver Carbon Dioxide Cell Line Cell Surface Proteins Cell surface Cells Cellular Morphology Centers of Research Excellence Central Nervous System Diseases Cerebrovascular system Cessation of life Chimeric Proteins Chinese Hamster Ovary Cell Chronic Chronic Phase Clinic Clinical Clinical Trials Collagen Collagen Type IV Communities Confidence Intervals Contralateral Control Groups Coupled Data Data Analyses Detection Development Devices Dextrans Diameter Disease Disease Progression Dose Drug Delivery Systems Drug Transport Electrical Resistance Endothelial Cells Endothelium Environment Enzyme-Linked Immunosorbent Assay Excision Exposure to Extracellular Matrix FDA approved Failure Fc Immunoglobulins Fibronectins Flow Cytometry Gel Generations Glass Glial Fibrillary Acidic Protein Glucose Growth Growth Factor Guide RNA Health Hour Housing Human Hyaluronic Acid Hydrogels Hypoxia Immunoglobulin G Immunoprecipitation In Situ Nick-End Labeling In Vitro Incubated Individual Infarction Injections Injury Insulin Receptor Intravenous Ipsilateral Ischemia Ischemic Stroke Label Laminin Lead Length Leukocytes Libraries Ligands Liposomes Lymphocyte Maintenance Mass Spectrum Analysis Measurement Measures Mediating Methods Microfluidic Microchips Microscopy Modeling Monitor Morphology Motor Mus Mutagens Names Nature Neurites Neuroglia Neurons Neuroprotective Agents Nutrient Nutrient availability Organ Outcome Oxygen PC12 Cells Patients Penetration Perfusion Permeability Persons Phage Display Pharmaceutical Preparations Phenotype Phosphorylation Physiological Plasmids Production Proteins Pump Quality of life Recovery Rehabilitation therapy Reperfusion Therapy Reporting Resected Rhodamine Rhodamine 123 Risk Rod Rodent Running Scheme Slide Source Specificity Stains Stroke Supporting Cell Surface System TFRC gene Tariquidar Techniques Testing Therapeutic Tight Junctions Time Tissue Sample Tissues Tracer Transfection Translating Traumatic Brain Injury Tubulin United States National Institutes of Health Universities Validation Vascular Endothelium Visualization West Virginia Western Blotting Work Yeasts antibody libraries antibody test aquaporin 4 blood-brain barrier crossing blood-brain barrier function candidate identification cell immortalization cell motility cell type chronic stroke cross reactivity density deprivation design design and construction disability disulfide bond efficacy testing experimental study fabrication field study human disease human model human tissue improved in vitro Model induced pluripotent stem cell inflammatory marker insight lead candidate live cell microscopy lucifer yellow monolayer mouse model neuroinflammation neuronal growth neuroprotection non-invasive imaging normoxia occludin overexpression placebo group polydimethylsiloxane power analysis preclinical study prevent protein folding receptor recruit response screening shear stress side effect small molecule therapeutics stem cell model stem cells stroke model stroke recovery stroke therapy success symptom management targeted treatment trafficking transcytosis translational therapeutics transport inhibitor uptake vector

项目摘要

Stroke is one of the leading causes of disability and death in the US. One of the contributing factors to this is the lack of FDA approved treatments for chronic stroke. Aside from tissue plasminogen activator (tPA), there are no other approved therapeutics with disease modifying effects; current therapy is entirely focused on symptom management and rehabilitation. This is in part due to the lack of understanding of the underlying mechanisms involved, as well as the difficulty of delivering therapeutics through the blood-brain barrier (BBB) and into the brain. This project aims to address both of these difficulties through the development of a human induced pluripotent stem cell (hiPSC) derived model of the BBB incorporating hiPSC derived brain microvascular endothelial like cells (BMECs), astrocytes, and neurons. Using a 3D microvessel platform ischemia will be modeled using perturbations in shear flow, oxygen concentration, and nutrient availability. The resultant phenotype of the cells will be investigated using live cell microscopy to measure permeability of fluorescent tracer molecules, as well as morphological changes in the cells. Using this model, as well as a simplified 2D model of ischemia, a library of antibodies will be screened for improved targeting and delivery to the human ischemic brain. Finally, the antibodies identified in the screen will be tested for efficacy in delivering known non-brain penetrant neuroprotectant growth factors to the ischemic brain both in the in vitro model and in a mouse model of stroke. In summary, this project will establish a human specific model of the BBB during ischemic stroke and identify antibodies that are able to specifically target and deliver therapeutics to the ischemic region of the brain.

中风是美国残疾和死亡的主要原因之一。造成这一现象的因素之一缺乏 FDA 批准的慢性中风治疗方法。除了组织纤溶酶原激活剂（tPA）之外，没有其他经批准具有疾病缓解作用的疗法；目前的治疗完全集中于关于症状管理和康复。造成这种情况的部分原因是缺乏对所涉及的潜在机制，以及通过血脑提供治疗的难度屏障（BBB）并进入大脑。该项目旨在通过开发人类诱导多能干细胞 (hiPSC) 衍生的 BBB 模型 hiPSC 衍生的脑微血管内皮样细胞 (BMEC)、星形胶质细胞和神经元。使用 3D 将使用剪切流、氧浓度、和营养物质的可用性。将使用活细胞显微镜研究所产生的细胞表型测量荧光示踪分子的渗透性以及细胞的形态变化。使用该模型以及简化的缺血二维模型，将筛选抗体库改善对人类缺血大脑的靶向和输送。最后，筛选出的抗体将测试将已知的非脑渗透性神经保护生长因子递送至体外模型和小鼠中风模型中的缺血性大脑。综上所述，本项目将建立缺血性中风期间 BBB 的人类特异性模型，并鉴定能够专门针对大脑缺血区域并向其提供治疗药物。