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

• 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
• 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

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 围产期动脉缺血性卒中很常见，发病率很高，长期严重 神经功能缺陷。围产期卒中导致显着的发病率和严重的长期神经系统损伤 和认知缺陷。超过一半的脑瘫儿童是足月出生的。它已经成为 明确受伤时大脑的成熟阶段在大脑模式中起着关键作用 对人类的损害，但围产期卒中不同病理生理学的潜在机制 对于成人中风的了解仍然知之甚少。脉络丛（CP）可以保护大脑 物理和生化障碍以及改变神经炎症。直到最近，CP 被忽视为潜在的治疗靶点，但现在被认为是治疗的药理学靶点 神经退行性疾病的治疗。我们将研究 CP 的神经保护作用 对抗在年龄适当的中风模型中诱发的新生儿中风，短暂性大脑中动脉 出生后第 9 天 (P9) 小鼠的闭塞 (tMCAO)。特别是，我们将确定如何 通过 CP 进入的单核细胞可以保护 Toll 样受体 2 (TLR2)（关键之一） 先天免疫系统的组成部分，在 CP 介导的神经保护中发挥核心作用。 我们假设单核细胞通过 CP 的运输可以保护新生儿大脑免受 tMCAO 侵害 以 TLR2 依赖的方式。 我们将确定通过 CP 进入的单核细胞表型与 新生小鼠 tMCAO 后的炎症和损伤 (Aim1) 并确定 TLR2- 的机制 通过损伤时功能性或功能失调的 CP 介导单核细胞运输（目标 2）。我们将用独特的 Cx3Cr1GFP/+/CCR2RF/P+ 小鼠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报告基因 luc/GFP-TLR2 小鼠进行非侵入性监测 活体受伤小鼠中 TLR2 实时上调的时空模式。我们的能力 通过弥散加权 MRI (DWI) 结合各种方法来识别 tMCAO 期间受伤的新生小鼠 在患有以下疾病的小鼠中进行生化检测，例如多色流式细胞术、EMSA 和细胞因子多重检测 CP 或/和 TLR2 的抑制功能将增强对 CP 保护作用的理解 新生儿脑卒中的屏障，为如何减轻急性新生儿脑卒中后的损伤提供新的见解 并确定血脑脊液屏障的药理学靶点。