A Vascularized Micro-Organ platform for the study of Brain-BBB-Blood interaction

用于研究脑-血脑屏障-血液相互作用的血管化微器官平台

• 批准号: 10512822
• 负责人: CHRISTOPHER C. W. HUGHES
• 金额: $ 65.31万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-03 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10512822
• 关键词: Acids; Affect; Alzheimer' s Disease; Amyloid beta-Protein; Anastomosis - action; Anatomy; Animals; Arteries; Astrocytes; Basement membrane; Biological Models; Blood; Blood - brain barrier anatomy; Blood Cells; Blood Glucose; Blood Substitutes; Blood Vessels; Blood flow; Brain; CADASIL; CAG repeat; Cell Communication; Cells; Communication Barriers; Complex; Data; Development; Diagnostic; Disease Progression; Disease model; Endothelial Cells; Face; Foot Process; Gene Expression; Genes; Genetic; Glucose; Goals; Human; Huntington Disease; Huntington gene; Huntington protein; Individual; Insulin; Light; Lipids; Membrane; Microfluidics; Microglia; Modeling; Movement; Multiple Sclerosis; Neurodegenerative Disorders; Neurons; Organ; Parkinson Disease; Pathogenesis; Pathology; Pericytes; Pharmaceutical Preparations; Phase; Phenotype; Physiological; Process; Proteins; Regulation; Role; Running; Science; Signal Transduction; Stroke; Testing; Thalamencephalon; Therapeutic; Tight Junctions; Transferrin; Translating; Traumatic Brain Injury; Veins; WNT Signaling Pathway; Work; base; brain cell; brain parenchyma; cell type; cerebral blood volume; density; drug development; foot; in vitro Model; mutant; nervous system disorder; neuropathology; neurovascular; neurovascular unit; novel; novel therapeutic intervention; organ on a chip; polyglutamine; pressure; relating to nervous system; response; side effect; transcytosis

PROJECT SUMMARY For too long, studies of the Blood-Brain Barrier (BBB) have ignored the blood component of this interface, focusing almost exclusively on the cells of the Neurovascular Unit (NVU). The goal of the FOA to which we are responding aims to change this: “The intent of this FOA is to stimulate the development of a new field of blood-based science by re-defining the neurovascular unit as a component of the blood-brain interface. This will facilitate development of human- based neurovascular-blood models to identify targets for diagnostics and regulation of the blood-brain interface…” The NVU is comprised of endothelial cells (EC), pericytes and astrocytes, and a complex basement membrane, which work together to severely limit the free movement of molecules from the blood into the brain parenchyma. In response to local signals during development BBB EC develop tight junctions and have very low rates of transcytosis. The side-effect of this is that access of potentially therapeutic drugs into the brain is also compromised. In this proposal we will build on our well-established human Vascularized Micro-Organ (VMO) platform to create a novel blood-brain interface model, the VMO-B. In this model a network of human microvessels anastomoses to microfluidic channels representing an artery and a vein and are induced to a BBB phenotype by Wnt signaling. The vessels are invested by pericytes and contacted by astrocyte foot-processes. Importantly, we will run a blood substitute – VMOBlood – through the vessels that will mimic the composition of blood, including protein and lipid content. We will then use the VMO-B to investigate the process of BBB breakdown in the pathogenesis of Huntington’s disease. We already have preliminary data suggesting that expression of mutant HTT protein in EC causes BBB deficits. We will investigate crosstalk between blood and the cells of the NVU, and how expression of mHTT in each cell type affects cell-cell communication and barrier function. In the R61 phase we will pursue three aims: Aim 1 Develop a stable MPS BBB model with perfused microvasculature; Aim 2 Incorporate flow of blood into BBB microfluidic model; and, Aim 3 Characterize key transporters at the blood-brain interface. In the R33 phase we will use this platform to examine the role of the blood-brain interface in the pathology of HD through an additional two aims: Aim 4 Test the hypothesis that expression of mHTT in EC disrupts transport across the BBB leading to changes in the neural micro-environment; and, Aim 5 Test the hypothesis that expression of mHTT disrupts multiple cell-to-cell interactions at the blood- brain interface. Completion of this project will not only shed light on the neuropathology of Huntington’s disease, but will also yield a platform ideally suited to drug development and investigating the role of the blood- brain interface in numerous neurological diseases including Alzheimer’s disease, Multiple Sclerosis, Parkinson’s disease, stroke, CADASIL, and traumatic brain injury.

项目摘要 长期以来，对血脑屏障（BBB）的研究忽略了该界面的血液成分， 几乎只专注于神经血管单元（NVU）的细胞。我们所在的FOA的目标 响应旨在改变这一点： “该FOA的目的是通过重新定义刺激基于血液科学的新领域的发展 神经血管单元是血脑界面的组成部分。这将促进人类的发展 基于神经血管血管模型，以识别诊断和调节血脑的靶标 界面...” NVU已完成内皮细胞（EC），周细胞和星形胶质细胞以及一个复杂的地下膜， 它们共同工作以严重限制分子从血液到脑实质的自由运动。 为了响应开发过程中的本地信号，BBB EC开发紧密的连接处，率很低 转胞病。这的副作用是，可能会进入大脑的潜在治疗药物也是 妥协。在此提案中，我们将建立在我们建立的人类血管化微孔管（VMO）的基础上 创建新型血脑接口模型VMO-B的平台。在这个模型中，人类网络 微分系数吻合到代表动脉和静脉的微流体通道，并被诱导为BBB Wnt信号传导的表型。这些船只由周细胞投资，并由星形胶质流程处理。 重要的是，我们将通过模仿组成的血管运行血液替代品 - vmoblood 血液，包括蛋白质和脂质含量。然后，我们将使用VMO-B研究BBB的过程 亨廷顿氏病发病机理的崩溃。我们已经有初步数据表明 EC中突变HTT蛋白的表达导致BBB缺乏。我们将调查血液之间的串扰 NVU的细胞以及MHTT在每种细胞类型中的表达如何影响细胞 - 细胞通信和屏障 功能。在R61阶段，我们将追求三个目标：AIM 1开发稳定的MPS BBB模型 微脉管系统； AIM 2将血液流入BBB微流体模型中；并且，AIM 3表征密钥 血脑界面处的转运蛋白。在R33阶段，我们将使用此平台来检查 HD病理中的血脑界面通过另外两个目的：目标4检验以下假设： MHTT在EC中的表达破坏了跨BBB的运输，导致神经微环境的变化； 并且AIM 5检验以下假设：MHTT的表达会破坏血液上多个细胞对细胞相互作用 脑接口。该项目的完成不仅会阐明亨廷顿的神经病理学 疾病，但也将产生一个非常适合药物开发和研究血液的作用的平台 许多神经系统疾病的脑接口，包括阿尔茨海默氏病，多发性硬化症，帕金森氏症 疾病，中风，卡达西尔和脑外伤。