Leukocyte trafficking through the choroid plexus as modulator of neonatal focal stroke

白细胞通过脉络丛的运输作为新生儿局灶性中风的调节剂

基本信息

批准号：
9188681
负责人：
Zinaida S Vexler
金额：
$ 23.19万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9188681
关键词：
Acute Adult Affect Age Agonist Animal Model Antibodies Biochemical Biological Assay Blood Blood - brain barrier anatomy Blood Circulation Blood Vessels Brain Brain Injuries Cells Cerebral Palsy Child Cognitive deficits Color Data Diffusion Magnetic Resonance Imaging EMSA Elderly Event Extravasation Family Flow Cytometry Functional disorder Homing Human Immune Immune system Immunofluorescence Immunologic Incidence Infant Inflammation Inflammatory Injury Ischemic Brain Injury Ischemic Stroke Knock-in Learning Leukocyte Trafficking Life Live Birth Mediating Microglia Middle Cerebral Artery Occlusion Modeling Molecular Monitor Morbidity - disease rate Movement Multiple Sclerosis Mus Neonatal Neurodegenerative Disorders Neurologic Pathology Pattern Perinatal Peripheral Phenotype Play Public Health Rattus Reporter Role Route Signal Transduction Site Societies Spinal cord injury Staging Stroke Structure Structure of choroid plexus Surgical sutures TLR2 gene Term Birth Time Up-Regulation abstracting bioluminescence imaging blood cerebrospinal fluid barrier brain parenchyma clinically relevant cost cytokine disability injured innovation insight mature animal monocyte mouse toll-like receptor 2 neglect neonatal brain neonatal stroke neonate neuroinflammation neuroprotection novel perinatal stroke postnatal protein expression pup targeted treatment tool trafficking

项目摘要

Abstract Perinatal arterial ischemic stroke is common and produces significant morbidity and severe long-term neurological deficits. Perinatal stroke leads to significant morbidity and severe long-term neurological and cognitive deficits. More than half of all children with cerebral palsy are born at term. It has become clear that the maturational stage of the brain at the time of injury plays a key role in the pattern of brain damage in humans but the underlying mechanisms of the differing pathophysiology of perinatal stroke and stroke in the adult are still poorly understood. The choroid plexuses (CPs) may protect the brain as physical and biochemical barriers and by modifying neuroinflammation. Until recently CPs were neglected as a potential target for therapy but are now considered as pharmacological target for treatment of neurodegenerative diseases. We will investigate the neuroprotective role of the CPs against neonatal stroke induced in an age-appropriate stroke model, a transient middle cerebral artery occlusion (tMCAO) in postnatal day 9 (P9) mice. In particular, we will determine how subsets of monocytes that enter through the CPs protect and whether toll-like receptor 2 (TLR2), one of key components of the innate immune system, plays a central role in CPs-mediated neuroprotection. We hypothesize that monocyte trafficking through CPs protects neonatal brain against tMCAO in a TLR2 dependent manner. We will determine relationships between the monocyte phenotypes entering through CPs and inflammation and injury after tMCAO in neonatal mice (Aim1) and define the mechanisms of TLR2- mediated monocyte trafficking via functional or dysfunctional CPs on injury (Aim 2). We will use unique Cx3Cr1GFP/+/CCR2RF/P+ mice to distinguish monocytes from microglia and characterize monocyte/microglial phenotypes and use double-reporter luc/GFP-TLR2 mice to non-invasively monitor the spatial-temporal pattern of TLR2 upregulation in real time in living injured mice. Our ability to identify injured neonatal mice during tMCAO by diffusion-weighted MRI (DWI) combined with various biochemical assays, such as multi-color flow cytometry, EMSA and cytokine multiplex, in mice with inhibited function of CPs or/and TLR2 will enhance the understanding of the role of CPs as protective barrier in neonatal stroke and provide new insights on how to alleviate injury after acute neonate stroke and identify pharmacological targets at the blood-CSF barrier.

抽象的围产期动脉缺血性中风很常见，并产生明显的发病率和严重的长期神经缺陷。围产期中风会导致明显的发病率和严重的长期神经系统和认知缺陷。所有脑瘫儿童中有一半以上出生于学期。它已经变成了清楚地表明，受伤时大脑的成熟阶段在大脑的模式中起着关键作用人类的损害，但围产期中风不同病理生理学的潜在机制成年人中的中风仍然很了解。脉络丛（CPS）可能会保护大脑物理和生化障碍，并通过修改神经炎症。直到最近CP 被忽视为治疗的潜在靶标，但现在被视为神经退行性疾病的治疗。我们将研究CP的神经保护作用针对适合年龄的中风模型引起的新生儿中风，这是一种短暂的中大脑动脉闭塞（TMCAO）在产后第9天（P9）小鼠中。特别是，我们将确定如何通过CPS保护输入的单核细胞以及Toll样受体2（TLR2）是否是关键之一先天免疫系统的成分在CPS介导的神经保护中起着核心作用。我们假设通过CPS进行单核细胞运输可保护新生儿大脑免受TMCAO的侵害以TLR2的方式。我们将确定通过CPS进入的单核细胞表型与 TMCAO在新生儿小鼠中的炎症和损伤（AIM1），并定义TLR2-的机制通过功能或功能失调的CPS介导的单核细胞运输（AIM 2）。我们将使用独特的 CX3CR1GFP/+/CCR2RF/P+小鼠，以区分单核细胞和小胶质细胞并表征单核细胞/小胶质细胞表型，并使用双重重复蛋白LUC/GFP-TLR2小鼠进行非侵入性监测在生命受伤的小鼠中实时上调TLR2的时空模式。我们的能力通过扩散加权MRI（DWI）与各种相结合，在TMCAO期间鉴定受伤的新生小鼠在具有的小鼠中 CPS或/和TLR2的抑制功能将增强对CP的作用的理解新生儿中风的障碍，并提供有关如何减轻急性新生儿中风后伤害的新见解并确定血液-CSF屏障的药理靶标。