Dynamic Circadian Regulation of the Blood-Brain Interface in a Human Brain-mimicking Microfluid Chip

模拟人脑微流体芯片中血脑界面的动态昼夜节律调节

• 批准号: 10912839
• 负责人: Martha U Gillette
• 金额: $ 48.03万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10912839
• 关键词: Address; Aging; Animal Model; Astrocytes; Award; Behavior; Biomimetics; Blood; Blood - brain barrier anatomy; Blood Coagulation Factor; Blood Vessels; Blood coagulation; Brain; Brain Diseases; Circadian Rhythms; Coagulation Process; Cognition Disorders; Consultations; Crying; Dementia; Deterioration; Development; Disease; Disease Management; Endothelial Cells; Endothelium; Engineering; Event; Exhibits; Extravasation; Fibrin; Fibrinogen; Gait; Genes; Goals; Grant; Health Promotion; Hemorrhage; Human; Hypertension; Incidence; Infection; Inflammation; Ischemic Stroke; Knowledge; Lead; Liquid substance; Microfluidics; Microglia; Modeling; Mus; Names; Nerve Degeneration; Neurons; Optics; Outcome; Pattern; Personal Satisfaction; Persons; Pharmacotherapy; Phase; Physiology; Plasma; Plasminogen Activator; Plasminogen Activator Inhibitor 1; Predisposition; Reporter; Reporter Genes; Reporting; Research; Role; Side; Sleep; Sleep Deprivation; Stress; Stroke; Stroke prevention; Syndrome; System; Testing; Therapeutic; Thrombin; Tight Junctions; Time; Transgenes; Traumatic Brain Injury; United States; Vascular Endothelial Cell; Vascular Endothelium; Venus; Wakefulness; brain parenchyma; brain tissue; cell injury; cerebral microbleeds; cerebrovascular; circadian; circadian pacemaker; circadian regulation; design; disorder prevention; glial activation; high reward; high risk; in vivo; induced pluripotent stem cell; inhibitor; innovation; insight; member; millimeter; mimicry; neuron loss; neurovascular; novel; pressure; public health relevance; response; stem cells; stroke incidence; tool; translational applications; translational model; vascular factor

Cerebral microbleeds (CMBs) and microhemorrhages (CMHs) result from blood leakage across the blood- brain interface (BBI). Subsequent millimeter-sized blood clots lead to inflammation, cellular injury, and neuro- degeneration. Such CMBs are associated with deterioration of BBI integrity with aging, disease, traumatic brain injury, and the sequelae of strokes, which impact >795,000 people in the United States every year. Notably, CMBs and hemorrhagic/ischemic stroke occurrence is not random, but rather clusters in early day or evening. Understanding of the circadian dynamics of the BBI with respect to vulnerability to blood leakage is limited. It has been difficult to study in vivo or via on-chip models and there is no drug treatment. In addressing this gap, this proposal responds to FOA RFA-HL-20-021. The purpose of this FOA is to support high risk/high reward research on the blood component of the Blood-Brain Barrier and the associated Interface to facilitate the development of a more complete neurovascular-blood model for translational applications with direct relevance to humans. It is an R61/R33 Exploratory /Developmental Phased Award. Because knowledge of the circadian dynamics in BBI vulnerability to blood leakage is limited, we aim to create a new biomimetic brain transport model with mimicry of the coagulation system and circadian rhythm. We will develop an innovative microfluidic platform to examine interactions of coagulation factors and circadian oscillations of both 1) the blood/vascular components and 2) dynamic vascular pressure across the BBI over the circadian cycle. We propose to reproduce circadian dynamics of the BBI by culturing human endothelial cells containing a clock-gene reporter on the ‘vascular’ side with polarized astrocytes, neurons, and microglia in the ‘brain’ compartment. This project assembles the expertise needed to facilitate the creation of enhanced platforms that more closely model the human BBI. Contributions of team members will be: Han–Microfluidics and biotransport analysis; Kong– engineering of BBI; Gillette–Assemble/validate a human iPSC circadian reporter-in-chip and assess rhythms and fluxes, and consultation from Flick on blood coagulation factors and Obrietan on the circadian reporter transgene. This grant will be separated into 2 phases: Focus in YR 1-2 (R61) will be on establishing tools and in YR 3-5 (R33) on utilizing those tools to achieve our research goals. This will enable us to replicate dynamics of the BBI in human brain and to probe it in the context of the oscillatory circadian cycle that drives integrative physiology and behavior, including sleep and wakefulness. By targeting both sides of the BBI and their intersection, we will gain insights into the emerging view that the BBI is plastic, changing with time-of-day, loss of sleep, the stress of infection, and aging. This has significant implications for the role of the circadian clock in blood coagulation in the brain and neurovascular function. The outcome will contribute to developing therapeutic opportunities that target the temporal occurrence of adverse cerebrovascular events, including hypertension, cognitive disorders and dementias, and gait syndromes.

脑微粒（CMB）和微毛发（CMHS）是由于血液渗漏而引起的 脑接口（BBI）。随后的毫米大小的血凝块导致感染，细胞损伤和神经 - 退化。这种CMB与BBI完整性与衰老，疾病，创伤性大脑相对重要有关 受伤和中风的后遗症，每年在美国影响> 795,000人。尤其， CMB和出血/缺血性中风的发生不是随机的，而是在早期甚至什至。 了解BBI在易受血液泄漏方面的昼夜动力学的理解是有限的。它 在体内或通过片上模型很难研究，没有药物治疗。在解决这个差距时， 该提案对FOA RFA-HL-20-021的响应。该FOA的目的是支持高风险/高奖励 研究血脑屏障的血液成分和相关界面，以促进 开发更完整的神经血管血管模型，用于具有直接相关性的翻译应用 给人类。这是R61 /R33探索 /发展分阶段奖。因为对昼夜候的知识 BBI对血液泄漏的脆​​弱性的动态有限，我们旨在创建新的仿生脑运输 模仿凝结系统和昼夜节律的模型。我们将开发创新的微流体 检查凝血因子和昼夜节律的相互作用的平台1）血液/血管 组件和2）在整个昼夜节律上跨BBI的动态血管压力。我们建议 通过文化人类内皮细胞复制BBI的昼夜节律动力学 在“大脑”室中具有极化星形胶质细胞，神经元和小胶质细胞的“血管”一侧。这个项目 组装促进创建增强平台所需的专业知识，该平台更紧密地建模 人BBI。团队成员的贡献将是：Han -Microfluidics和Biotransport分析；孔 - BBI的工程；吉列 - 组装/验证人类IPSC昼夜节目记者和评估节奏 和通量，以及Flick对血液凝结因素的咨询和昼夜节目记者的Obrietan 转基因。该赠款将分为2个阶段：重点1-2（R61）将建立工具和 在YR 3-5（R33）中，使用这些工具来实现我们的研究目标。这将使我们能够复制动态 BBI在人脑中的BBI，并在驱动整合的振荡性昼夜节律的背景下进行探测 生理和行为，包括睡眠和清醒。通过针对BBI及其两侧 交叉点，我们将深入了解BBI是塑料的新兴观点，随着时间的流逝而变化 睡眠，感染的压力和衰老。这对昼夜节律在 大脑和神经血管功能的血液凝血。结果将有助于发展 针对暂时发生不良脑血管事件的治疗机会，包括 高血压，认知障碍和痴呆症以及步态综合征。