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

• 批准号: 9286282
• 负责人: Thomas Neumann
• 金额: $ 70.09万
• 依托单位: NORTIS, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2018-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9286282
• 关键词: Acquired Immunodeficiency Syndrome; Alzheimer' 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 Targeting; Drug Transport; Endothelial Cells; Failure; Grant; Health; Human; IL6 gene; Label; Leukocytes; Life; Lipopolysaccharides; Liquid substance; Malignant Neoplasms; Mannitol; Marketing; 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; Qualifying; 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; in vitro Assay; in vitro Model; in vivo; migration; mouse model; occludin; predict clinical outcome; protein expression; research study; response; self assembly; success; trafficking

 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（由适用提供）：血脑屏障（BBB）是由微血管和毛细血管形成的紧密屏障，可控制营养素，液体，代谢产物以及血液与大脑之间的药物。 BBB的功能受损参与了折磨大脑的许多主要病理，例如阿尔茨海默氏病，多发性硬化症，帕金森氏病，艾滋病，中风和癌症的大脑表现。尽管神经治疗领域是制药行业中增长最大，增长最快的市场之一，但目前由于缺乏Reliabl预测体内BBB渗透性的体外评估而损害了进步。现有的模型均未充分复制BBB的有机微环境，其中脑内皮细胞（EC），周细胞（PC）和星形胶质细胞（ACS）以特征性结构布置。拟议的工作利用Nortis，Inc。最近开发的芯片技术来创建一次性微流体芯片中的3D组织微环境。芯片设计使生活，灌注的微脉管系统可以整合，使其适合研究屏障功能。令人惊讶的是，广泛的初步数据表明，人脑EC，PC和ACS具有自组装成诺蒂斯芯片内BBB样结构的能力。这些数据将被利用以进一步发展，并最终将小鼠和人类的BBB模型商业化。第一阶段的目的是实现复制小鼠大脑关键BBB功能的模型。将开发和优化鼠标模型，以实现生存力，结构和功能。将测量紧密结（TJ）蛋白的表达（TJ）蛋白和转运蛋白P-糖蛋白是BBB的重要功能特征。微血管通透性将通过荧光标记的分子灌注来评估（AIM 1）。然后，该模型将通过屏障调节化合物脂多糖（LPS）的挑战，并评估TJ蛋白表达，分子渗透率和白细胞跨内皮迁移的相关变化（AIM 2）。在II期期间，小鼠BBB芯片将用于开发和限定特定的BBB测定法，例如转铁蛋白受体转运蛋白转运蛋白的活性，与LPS的BBB渗透性挑战以及刺激诱导的白细胞传播（AIM 1）。成功标准是z≥0.2的测定鲁棒性。 II期的目标2是开发人类BBB模型。人类模型将被优化，以概括BBB的关键结构和功能特征，包括TJ形成，渗透性和转运蛋白活性。为了证明效用，该模型将用LPS，甘露醇和血管紧张素II处理，并评估BBB结构和功能的相关变化。这些化合物中的每一种都有临床相关性，但通过一种不同的机制起作用。 AIM 3是符合特定的人类BBB测定法，并与临床数据建立相关性。在该赠款支持下开发的产品将显着增强基本，翻译和临床神经科学研究的进展，并将大大推动许多毁灭性疾病的治疗。