A microfluidic platform for modeling drug transport and cell trafficking across the blood-brain barrier

用于模拟药物跨血脑屏障转运和细胞运输的微流体平台

基本信息

批准号：
9356329
负责人：
Thomas Neumann
金额：
$ 68.77万
依托单位：
NORTIS, INC.
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：
2015-09-30 至 2019-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9356329
关键词：
Acquired Immunodeficiency Syndrome Alzheimer&apos s Disease Angiotensin II Animal Testing Architecture Astrocytes Biological Assay Blood Blood - brain barrier anatomy Blood capillaries Brain Cell Polarity Cell Survival Cells Characteristics Clinical Clinical Data Data Dextrans Disease Drug Industry Drug Modelings Drug Targeting Drug Transport Endothelial Cells Failure Grant Human IL6 gene Impairment Label Leukocytes Lipopolysaccharides Liquid substance Malignant Neoplasms Mannitol Measures Mediating Metabolic Microfluidic Microchips Microfluidics Modeling Multiple Sclerosis Mus Neurosciences Research Nutrient Organ P-Glycoprotein Parkinson Disease Pathology Perfusion Pericytes Peripheral Blood Mononuclear Cell Permeability Pharmaceutical Preparations Phase Proteins Small Business Innovation Research Grant Stimulus Stroke Structure Supporting Cell TFRC gene Technology Tight Junctions Tissue Engineering Tissues Transferrin White Blood Cell Count procedure Work brain endothelial cell capillary cell type clinically relevant cytokine density design experimental study in vitro Assay in vitro Model in vivo migration mouse model occludin predict clinical outcome protein expression public health relevance response self assembly success trafficking

项目摘要

DESCRIPTION (provided by applicant): The blood-brain barrier (BBB) is a tight barrier formed by microvessels and capillaries that control the passage of nutrients, fluids, metabolic products, and drugs between the blood and the brain. Impaired function of the BBB is involved in a number of major pathologies afflicting the brain, such as Alzheimer's disease, multiple sclerosis, Parkinson's disease, brain manifestations of AIDS, stroke, and cancer. Although the neurotherapeutics sector is among the largest and fastest growing markets in the pharmaceutical industry, progress is currently impaired by the lack of in vitro assays that reliabl predict in vivo BBB permeability. None of the existing models adequately replicates the organotypic microenvironment of the BBB, in which brain endothelial cells (ECs), pericytes (PCs) and astrocytes (ACs) are arranged in a characteristic architecture. The proposed work utilizes organ-on-chip technology recently developed by Nortis, Inc. for creating 3D tissue microenvironments in disposable microfluidic chips. The chip design enables the integration of living, lumenally perfused microvasculature, making it suitable for studying barrier function. Strikingly, extensive preliminary data indicate that human brain ECs, PCs, and ACs have the capacity to self-assemble into a BBB-like architecture within the Nortis chip. This data will be leveraged to further develop and eventually commercialize BBB models of mouse and human. The objective of Phase I is to achieve a model that replicates critical BBB functions of the mouse brain. The mouse model will be developed and optimized for viability, structure, and function. Expression of tight-junction (TJ) proteins and the transporter P-glycoprotein, an important functional characteristic of the BBB, will be measured. Microvessel permeability will be assessed by perfusion with fluorescently labelled molecules (Aim 1). The model will then be challenged with the barrier-modulating compound lipopolysaccharide (LPS), and evaluated for associated changes in TJ protein expression, molecule permeability, and leukocyte transendothelial migration (Aim 2). During Phase II, the mouse BBB chip will be used to develop and qualify specific BBB assays, such as transferrin receptor transporter activity, BBB permeability challenge with LPS, and stimuli-induced leukocyte transmigration (Aim 1). Success criteria is an assay robustness of Z' ≥ 0.2. Aim 2 of Phase II is to develop a human BBB model. The human model will be optimized to recapitulate key structural and functional features of the BBB, including TJ formation, permeability, and transporter activity. To demonstrate utility, the model will be treated with LPS, mannitol, and angiotensin II and evaluated for associated changes in BBB structure and function. Each of these compounds has clinical relevance but acts by a different mechanism. Aim 3 is to qualify specific human BBB assays and establish relevance to clinical data. The products developed with support from this grant will significantly enhance progress in basic, translational, and clinical neuroscience research and will significantly advance therapy for numerous devastating diseases.

描述：血脑屏障（BBB）是一个紧密的屏障，由微毛和毛s骨形成，而thassage的营养素，液体，代谢产物以及血液和大脑之间的药物涉及许多主要的病理。正如阿尔茨海默氏症（Alzheimer），多个扇贝病，大脑Athough神经疗法部门是最大，最快的市场工业Y，目前由于缺乏Reliabl预测体内BBB渗透率的体外测定而损害了进展。在特征性的结构中，脑内皮细胞（EC），周围的星形胶质细胞（PC）和星形胶质细胞（AC）。 Ture，适合研究屏障功能。将小鼠和人类的BBB模型进行商业化。 p-糖蛋白是BBB的重要特征，微分离的通透性将由荧光标记的分子散布（AIM 1）。渗透性和白细胞跨内皮迁移（AIM 2）。 AM 1）。甘露醇和血管紧张素II和对BBB结构和功能相关的相关变化的评估。将大大改进许多毁灭性疾病的治疗。