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， 几乎完全集中在神经血管单位（NVU）的细胞上。我们所要达到的FOA的目标 回应旨在改变这一点： “本FOA的目的是通过重新定义血液科学的新领域， 神经血管单位作为血脑界面的组成部分。这将有助于人类的发展- 基于神经血管血液模型，以确定诊断和调节血脑的目标 接口” NVU由内皮细胞（EC）、周细胞和星形胶质细胞以及复杂的基底膜组成， 它们共同作用，严重限制了分子从血液进入脑实质的自由运动。 在发育过程中，BBB EC对局部信号作出反应，形成紧密连接，并且具有非常低的细胞凋亡率。 转胞吞作用其副作用是，潜在的治疗药物进入大脑的通道也被破坏。 暴露了在这项提案中，我们将建立在我们完善的人类血管化微器官（VMO）的基础上。 平台创建一个新的血脑界面模型，VMO-B。在这个模型中， 微血管扩张到代表动脉和静脉的微流体通道，并被诱导到BBB 表型通过Wnt信号传导。血管由周细胞包裹，并与星形胶质细胞足突接触。 重要的是，我们将运行血液替代品- VMOBlood -通过血管，将模拟的成分， 血液，包括蛋白质和脂质含量。然后我们将使用VMO-B来研究BBB的过程 亨廷顿病的发病机制。我们已经有初步数据表明， 突变HTT蛋白在EC中的表达导致BBB缺陷。我们将研究血液和 NVU的细胞，以及mHTT在每种细胞类型中的表达如何影响细胞间通讯和屏障 功能在R61阶段，我们将追求三个目标：目标1开发一个稳定的MPS BBB模型， 微血管系统;目的2将血流纳入BBB微流体模型;以及，目的3表征关键 血脑界面的转运蛋白在R33阶段，我们将使用此平台来检查 血脑界面在HD病理学中的作用，通过另外两个目的：目的4检验假设， EC中mHTT的表达破坏了通过BBB的运输，导致神经微环境的变化; 目的5检验mHTT的表达破坏血液中多种细胞间相互作用的假设， 大脑接口这个项目的完成不仅将阐明亨廷顿舞蹈症的神经病理学， 疾病，但也将产生一个非常适合药物开发和研究血液作用的平台- 脑接口在许多神经系统疾病，包括阿尔茨海默病，多发性硬化症，帕金森氏症 疾病、中风、CADASIL和创伤性脑损伤。