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.

摘要 围产期动脉缺血性中风是常见的，并产生显着的发病率和严重的长期 神经缺陷围产期中风导致显著的发病率和严重的长期神经系统疾病 和认知缺陷超过一半的脑瘫患儿是足月出生的。它已成为 显然，受伤时大脑的成熟阶段在大脑的模式中起着关键作用。 但是围产期中风不同病理生理学的潜在机制 和中风的发病机制仍然知之甚少脉络丛（CP）可以保护大脑， 物理和生化屏障以及通过修饰神经炎症。直到最近，CP 被忽视为治疗的潜在靶点，但现在被认为是 神经退行性疾病的治疗。我们将研究CP的神经保护作用 针对在年龄适当的中风模型中诱导的新生儿中风，短暂的大脑中动脉 在出生后第9天（P9）小鼠中，观察到脑缺血（tMCAO）。特别是，我们将确定 通过CP进入的单核细胞是否保护了Toll样受体2（TLR 2）， 天然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运输保护新生儿脑免受tMCAO的影响 以TLR 2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（Aim 1）tMCAO后的炎症和损伤，并确定TLR 2- 介导的单核细胞运输通过功能或功能障碍的CP损伤（目的2）。我们将使用独特的 Cx 3Cr 1GFP/+/CCR 2 RF/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luc/GFP-TLR 2小鼠进行非侵入性监测 TLR 2在活体损伤小鼠中真实的时间上调的时空模式。的能力 通过弥散加权MRI（DWI）结合各种方法识别tMCAO期间受损的新生小鼠 生物化学测定，如多色流式细胞术，EMSA和细胞因子多重，在小鼠中， CPs或/和TLR 2的抑制功能将增强对CPs作为保护性细胞的作用的理解。 并为如何减轻急性新生儿卒中后的损伤提供新的见解 并确定血液-CSF屏障的药理学靶点。