Exosomes as the mechanism of mesenchymal stem cell brain repair in neonatal stroke

外泌体作为间充质干细胞脑修复新生儿中风的机制

• 批准号: 10373763
• 负责人: Zinaida S Vexler
• 金额: $ 44.41万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373763
• 关键词: Acute; Adult; Affect; Age; Behavior; Blood - brain barrier anatomy; Brain; Brain Hypoxia-Ischemia; Brain Injuries; Cell Communication; Cell Therapy; Cells; Cerebral Palsy; Cognitive deficits; Communication; Contralateral; Data; Development; Disease; Disease model; Elderly; Experimental Models; Failure; Family; Flow Cytometry; Fractionation; Histologic; Human; Infant; Infarction; Inflammation; Injury; Intervention; Intranasal Administration; Leukocyte Trafficking; Life; Literature; Live Birth; Long-Term Effects; Mediating; Mesenchymal Stem Cells; Methodology; Microglia; Middle Cerebral Artery Occlusion; Modeling; Mus; Neonatal; Nerve Degeneration; Nervous System Physiology; Neurodegenerative Disorders; Neurodevelopmental Disability; Neurologic Deficit; Neurons; Oligodendroglia; Organism; Outcome; Perinatal; Perinatal Brain Injury; Pharmacology; Play; Proteins; Rattus; Recovery; Reperfusion Therapy; Reporting; Research; Resolution; Rodent; Role; Sensorimotor functions; Severities; Signal Transduction; Societies; Source; Spinal cord injury; Stroke; Structure of choroid plexus; Term Birth; Testing; Therapeutic; Time; Traumatic Brain Injury; Vascular remodeling; Vesicle; base; brain repair; cell type; chemokine; clinically relevant; cost; cytokine; disability; effective therapy; exosome; extracellular vesicles; functional outcomes; improved; injured; injury and repair; innovation; macrophage; microvesicles; myelination; neonatal brain; neonatal hypoxic-ischemic brain injury; neonatal mice; neonatal stroke; neurobehavior; neuroinflammation; neuron loss; neuroprotection; new therapeutic target; non-invasive imaging; novel; novel therapeutic intervention; perinatal ischemic stroke; post stroke; postnatal; preservation; pup; repaired; restoration; sex; sham surgery; stem cell exosomes; stem cell survival; stroke clinical trials; stroke model; therapeutically effective; tool; uptake; white matter

Abstract Neonatal (perinatal) arterial ischemic stroke is a major cause of long-term neurological and cognitive deficits, including cerebral palsy and neurodevelopmental disabilities. While neonatal stroke is as common as in the elderly, literature has emerged that the stage of brain development at the time of stroke has a major impact on the pathophysiological mechanisms of brain damage. Previous therapeutic efforts were mostly focused on protecting neurons acutely, but such strategies appeared to be short-range. We reported that delayed intranasal administration of mesenchymal stem cells (MSC) protects the white matter and improves long-term functional outcomes in an experimental model of a transient middle cerebral artery occlusion (tMCAO) in neonatal rats. Extracellular vesicles (EV) are now believed to play fundamental role in cell-cell communication without direct cell-cell contacts in healthy and diseased organism and that EV is a part of neurodegenerative scenarios. Based on our preliminary data that exosomes released from MSC (MSC-exo) protect neonatal brain following subacute stroke, in this proposal we hypothesize that MSC-exo is the underlying mechanism of MSC-induced acute neuroprotection and long-term recovery after neonatal stroke via modulation of microglial cell signaling. Given that inflammation is a hallmark of perinatal brain injury, affecting both early injury and brain repair and connectivity later in life, and that microglial cells contribute to neuro- and vasoprotection in neonatal stroke, we will determine how uptake of untranasally administered MSC-exo by activated microglia/macrophages in ischemic-reperfused regions affects neuroinflammation and injury in neonatal mice of both sexes subjected to tMCAO and whether MSC-exo alter brain microenvironment via release of microvesicles and small EV from microglia (Aim 1), and determine the long-term effects of MSC-exo administration on myelination, brain repair and functional outcomes (Aim 2). To understand the mechanistic role of MSC-exo and their therapeutic potential for neonatal stroke, we will utilize state- of-the art experimental tools, including a clinically relevant perinatal focal arterial stroke model that we invented, in conjunction with pharmacological approaches and advanced non-invasive imaging methodologies (NanoSight, super resolution flow cytometry Alexa) and characterization of large/small EV and their “cargo” released from microglia from injured regions. The significance and novelty of the proposed studies are in advancing the mechanistic understanding of MSC-exo-induced cell-type specific effects in neonatal brain after stroke and identifying novel therapeutic targets to create effective and safe therapy for neonatal stroke.

抽象的 新生儿（围产期）动脉缺血性中风是长期神经和认知障碍的主要原因 缺陷，包括脑瘫和神经发育障碍，而新生儿中风也是如此。 与老年人一样常见，有文献表明，大脑发育阶段 中风对脑损伤的病理生理机制有重大影响。 治疗努力主要集中在急性保护神经元上，但这种策略似乎 我们报道了间充质干细胞（MSC）的鼻内给药。 在实验模型中保护白质并改善长期功能结果 新生大鼠短暂性大脑中动脉闭塞（tMCAO）现已成为细胞外囊泡（EV）。 据信在健康细胞之间无需直接细胞接触的情况下，在细胞间通讯中发挥基本作用 根据我们的初步判断，EV 是神经退行性病变的一部分。 数据显示，MSC 释放的外泌体 (MSC-exo) 可保护亚急性中风后的新生儿大脑 这个建议我们认为 MSC-exo 是 MSC 诱导的急性 通过调节小胶质细胞信号传导来实现新生儿中风后的神经保护和长期恢复。 鉴于炎症是围产期脑损伤的标志，都会影响早期损伤和脑修复 和以后生命中的连通性，小胶质细胞有助于新生儿的神经和血管保护 中风时，我们将确定活化的 MSC-exo 如何摄取非经鼻给药的 MSC-exo 缺血再灌注区域的小胶质细胞/巨噬细胞影响新生儿的神经炎症和损伤 接受 tMCAO 的男女小鼠以及 MSC-exo 是否通过改变大脑微环境 小胶质细胞释放微泡和小 EV（目标 1），并确定其长期影响 MSC-exo 给药对髓鞘形成、大脑修复和功能结果的影响（目标 2）。 MSC-exo 的机制作用及其对新生儿中风的治疗潜力，我们将利用状态- 最先进的实验工具，包括我们开发的临床相关的围产期局灶性动脉卒中模型 结合药理学方法和先进的非侵入性成像发明 方法（NanoSight、超分辨率流式细胞术 Alexa）和大/小特征 EV 及其从受伤区域的小胶质细胞释放的“货物”的意义和新颖性。 拟议的研究旨在推进对 MSC-exo 诱导的细胞类型的机制理解 中风后新生儿大脑的具体影响，并确定新的治疗靶点以创造有效的 以及新生儿中风的安全治疗。