The role of neurovascular interactions in the development and regulation of the blood-brain barrier

神经血管相互作用在血脑屏障发育和调节中的作用

• 批准号: 10472652
• 负责人: Natasha O'Brown
• 金额: $ 12.93万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-20 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10472652
• 关键词: 4D Imaging; Address; Alzheimer' s Disease; Area; Basic Science; Behavior; Behavioral; Behavioral Assay; Binding; Biological Assay; Biological Models; Biology; Blood - brain barrier anatomy; Blood Vessels; Brain; Cell Transplantation; Cell physiology; Cells; Cellular biology; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Confocal Microscopy; Defect; Development; Developmental Biology; Drug Delivery Systems; Drug Screening; Embryo; Endothelial Cells; Environment; Exhibits; Extravasation; Faculty; Fishes; Foundations; Functional disorder; Future; Genes; Genetic; Genomics; Goals; Homeostasis; Human; Image; Image Analysis; Knock-out; Laboratories; Larva; Leadership; Learning; Maintenance; Malignant neoplasm of brain; Maps; Measures; Mediator of activation protein; Mentors; Mesoderm; Midbrain structure; Missense Mutation; Molecular; Molecular Biology; Mus; Mutagenesis; Neural Crest; Neuraxis; Neurodegenerative Disorders; Neurons; Neurophysiology - biologic function; Parkinson Disease; Pericytes; Permeability; Pharmaceutical Preparations; Phenotype; Physiology; Population; Positioning Attribute; Property; Prosencephalon; Proteoglycan; Regulation; Reporter; Research; Resolution; Role; Signal Transduction; System; Techniques; Tertiary Protein Structure; Testing; Therapeutic; Tracer; Training; Transgenic Organisms; Vascular Endothelial Cell; Vertebrates; Work; Zebrafish; base; behavior change; behavioral response; blood-brain barrier function; blood-brain barrier permeabilization; brain cell; brain endothelial cell; career; cell type; developmental genetics; expectation; experimental study; high resolution imaging; in vivo; innovation; insight; medical schools; mouse model; mutant; nervous system disorder; neurotransmission; neurovascular; neurovascular unit; new therapeutic target; novel; novel therapeutics; oligodendrocyte precursor; precursor cell; quantitative imaging; reflectance confocal microscopy; response; screening; seal; skills; small molecule; spatiotemporal; therapeutic target; transcriptome sequencing; zebrafish development

Project Summary The blood-brain barrier (BBB) maintains homeostasis of the central nervous system (CNS) chemical environment, and is critical for proper neural function; BBB breakdown has been implicated in neurodegenerative diseases, including Alzheimer’s disease and Parkinson’s disease. The BBB is also a huge obstacle for drug delivery to the brain. For both of these reasons, we need a better understanding of the molecular regulators of BBB development and function. This K99/R00 proposal will support the PI, Dr. Natasha O’Brown, in developing innovative strategies and acquiring new skill sets to study the signaling between neurovascular cells and blood vessels involved in establishing BBB function. The experiments will be initiated during the mentored period (carried out in Dr. Sean Megason’s lab at Harvard Medical School) and continue in the PI’s own laboratory upon obtaining an independent position. Dr. O’Brown’s long-term career goal is to unravel the cellular and molecular interactions that direct BBB development and maintenance, with the hope of identifying new targets for therapeutic drug delivery into the brain. Dr. O’Brown’s previous work identified a novel zebrafish mutant with vastly increased BBB permeability. The leaky phenotype mapped to a missense mutation in the neuronally-produced secreted proteoglycan Spock1. Spock1 is the first identified neuronal factor that establishes BBB properties, and it therefore represents a huge opportunity to learn more about how endothelial cell permeability is regulated by the CNS microenvironment. Aim 1 will use zebrafish genetics, cell transplantation and tracer leakage assays to perturb and assess Spock1’s function in regulating BBB properties. Aim 2 will use a combination of lineage tracing, live imaging and RNA sequencing to characterize how loss of Spock1 alters CNS cellular interactions leading to increased BBB leakage. Finally, Aim 3 will establish a connection between BBB dysfunction and altered behavior, generating a new platform for a small molecule screen to identify novel molecules that alter BBB function. The discovery of novel molecular regulators of BBB function will form the foundation of Dr. O’Brown’s independent laboratory. Dr. O’Brown’s extensive training in developmental biology and genetics makes her an ideal candidate to further develop this important research area. Building on this training, she will develop expertise in zebrafish genetic and cellular manipulations, 4D high-resolution live imaging and analysis, and in performing and analyzing behavioral drug screens. She will be mentored throughout the K99 period by Drs. Sean Megason, Leonard Zon, Chenghua Gu and Randall Peterson, world-class experts in zebrafish development and quantitative imaging, zebrafish genetics and genomics, BBB physiology and small molecule screens. Together, the proposed experiments and training will position Dr. O’Brown to be an excellent candidate for independent faculty positions, and for a leadership role in research into the cellular and molecular control of BBB development and function.

项目概要 血脑屏障 (BBB) 维持中枢神经系统 (CNS) 化学物质的稳态 环境，对于正常的神经功能至关重要； BBB 崩溃与神经退行性疾病有关 疾病，包括阿尔茨海默病和帕金森病。 BBB也是药物的巨大障碍 输送到大脑。由于这两个原因，我们需要更好地了解分子调节剂 BBB 的发育和功能。该 K99/R00 提案将支持 PI Natasha O’Brown 博士开发 创新策略和获取新技能来研究神经血管细胞和血液之间的信号传导 参与建立 BBB 功能的血管。实验将在指导期间启动 （在哈佛医学院 Sean Megason 博士的实验室中进行）并在 PI 自己的实验室中继续进行 获得独立地位。奥布朗博士的长期职业目标是解开细胞和分子的谜团 指导 BBB 开发和维护的互动，希望能够确定新的目标 治疗药物输送到大脑中。 奥布朗博士之前的工作发现了一种新型斑马鱼突变体，其血脑屏障通透性大大增加。这 渗漏表型映射到神经元产生的分泌蛋白聚糖 Spock1 的错义突变。 Spock1 是第一个被识别的建立 BBB 特性的神经元因子，因此它代表了一个巨大的 有机会更多地了解中枢神经系统微环境如何调节内皮细胞通透性。 目标 1 将使用斑马鱼遗传学、细胞移植和示踪剂泄漏测定来干扰和评估 Spock1 具有调节 BBB 特性的功能。目标 2 将结合谱系追踪、实时成像和 RNA 测序来表征 Spock1 的缺失如何改变 CNS 细胞相互作用，从而导致 BBB 增加 泄漏。最后，目标 3 将在 BBB 功能障碍和行为改变之间建立联系，从而产生 用于小分子筛选的新平台，用于识别改变 BBB 功能的新分子。的发现 血脑屏障功能的新型分子调节剂将构成奥布朗博士独立实验室的基础。 奥布朗博士在发育生物学和遗传学方面接受过广泛的培训，使她成为进一步发展的理想人选 发展这一重要的研究领域。在这次培训的基础上，她将发展斑马鱼遗传方面的专业知识 和细胞操作、4D 高分辨率实时成像和分析以及执行和分析 行为药物筛查。她将在整个 K99 期间接受 Drs. 的指导。肖恩·梅加森、伦纳德·佐恩、 顾成华和兰德尔·彼得森是斑马鱼发育和定量成像领域的世界级专家， 斑马鱼遗传学和基因组学、BBB 生理学和小分子筛选。共同提出的 实验和培训将使奥布朗博士成为独立教职职位的优秀候选人， 并在 BBB 发育和功能的细胞和分子控制研究中发挥领导作用。