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: 10915164
• 负责人: Natasha O'Brown
• 金额: $ 24.03万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10915164
• 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; Blood brain barrier dysfunction; Brain; Cell Transplantation; Cell physiology; Cells; Cellular biology; Central Nervous System; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Confocal Microscopy; Defect; Development; Developmental Biology; Dimensions; Drug Delivery Systems; Drug Screening; Embryo; Endothelial Cells; Environment; Exclusion; Exhibits; Extravasation; Faculty; Fishes; Foundations; Future; Genes; Genetic; Genomics; Goals; Homeostasis; Human; Image; Image Analysis; Knock-out; Laboratories; Larva; Leadership; Learning; Maintenance; Malignant neoplasm of brain; Maps; Measures; Mediator; Mentors; Mesoderm; Midbrain structure; Missense Mutation; Molecular; Molecular Biology; Mus; Mutagenesis; Neural Crest; 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; 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.

项目摘要 血脑屏障（BBB）维持中枢神经系统（CNS）化学物质的稳态， BBB破坏与神经退行性疾病有关，并且对适当的神经功能至关重要; 疾病，包括阿尔茨海默病和帕金森病。血脑屏障也是药物治疗的巨大障碍 输送到大脑。由于这两个原因，我们需要更好地了解的分子调节剂， BBB的发育和功能。本K99/R 00提案将支持PI Natasha O 'Brown博士开发 创新策略和获得新的技能，以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。实验将在辅导期间启动 （在哈佛医学院Sean Megason博士的实验室进行），并在PI自己的实验室继续进行， 获得独立地位。O 'Brown博士的长期职业目标是解开细胞和分子 相互作用，指导BBB的发展和维护，希望确定新的目标， 将治疗药物输送到大脑中。 博士O 'Brown之前的工作确定了一种新的斑马鱼突变体，其BBB通透性大大增加。的 渗漏表型映射到神经元产生的分泌蛋白聚糖Spock 1中的错义突变。 Spock 1是第一个确定的建立BBB特性的神经元因子，因此它代表了一个巨大的 有机会了解更多关于内皮细胞通透性是如何调节中枢神经系统微环境。 目的1将使用斑马鱼遗传学，细胞移植和示踪剂泄漏试验来干扰和评估Spock 1的 在调节血脑屏障特性中的作用。目标2将使用谱系追踪，活体成像和RNA的组合 测序以表征Spock 1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将建立BBB功能障碍和行为改变之间的联系， 小分子筛选的新平台，以确定改变BBB功能的新分子。的发现 BBB功能的新型分子调节剂将成为O 'Brown博士独立实验室的基础。 博士奥布朗在发育生物学和遗传学方面的广泛培训使她成为进一步发展的理想候选人 开发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学方面的专业知识 和细胞操作，4D高分辨率实时成像和分析，以及在执行和分析 行为药物筛查在整个K99期间，她将接受Sean Megason、伦纳德·Zon博士的指导 Chenghua Gu和Randall Peterson是世界级的斑马鱼发育和定量成像专家， 斑马鱼遗传学和基因组学、血脑屏障生理学和小分子筛选。在一起，拟议的 实验和培训将使O 'Brown博士成为独立教师职位的优秀候选人， 并在研究BBB发育和功能的细胞和分子控制方面发挥领导作用。