Mechanisms underlying extracellular vesicle mediated changes in inflammation, neural circuitry and plasticity following cortical injury in aged monkeys

细胞外囊泡介导老年猴子皮质损伤后炎症、神经回路和可塑性变化的机制

• 批准号: 10501439
• 负责人: TARA L MOORE
• 金额: $ 68.05万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10501439
• 关键词: Acute; Alzheimer' s disease related dementia; Anti-Inflammatory Agents; Area; Astrocytes; Attention; Automobile Driving; Axon; Behavioral; Biological Markers; Blood; Brain; Cell Nucleus; Cell Survival; Cells; Chronic; Cognitive deficits; Craniocerebral Trauma; Development; Early treatment; Encapsulated; Environment; Exhibits; FDA approved; FOS gene; Female; Glucose; Hand; Harvest; Histologic; Hour; Human; Immediate-Early Genes; Immune signaling; In Vitro; Inflammation; Inflammatory; Injury; Intravenous; Investigation; Knowledge; Label; Lesion; Macaca mulatta; Magnetic Resonance Imaging; Maps; Measures; Mediating; Mesenchymal Stem Cells; MicroRNAs; Microglia; Microscopy; Molecular; Molecular Mechanisms of Action; Monkeys; Motor; Motor Cortex; Myelin; Nature; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neuroglia; Neuronal Plasticity; Neurons; Oligodendroglia; Oxidative Stress; Oxygen; Phenotype; Physiological; Physiology; Postoperative Period; Process; Proteins; Proteomics; Recovery; Recovery Support; Recovery of Function; Resolution; Signaling Protein; Stroke; Synapses; Testing; Therapeutic; Time; Tissue Harvesting; Tracer; Training; Validation; Work; age related; aged; aging brain; anatomical tracing; behavioral outcome; brain cell; brain magnetic resonance imaging; brain tissue; cell injury; cell type; cohort; deprivation; excitotoxicity; extracellular vesicles; hand dysfunction; in vitro Model; in vivo; inflammatory marker; injury recovery; male; mesenchymal stromal cell; motor deficit; motor function recovery; multidisciplinary; multimodality; myelination; neural circuit; neuroinflammation; neuronal excitability; neuroprotection; oxidative damage; patch clamp; relating to nervous system; remyelination; repaired; response; response to injury; sex; single-cell RNA sequencing; tissue repair; transcriptome sequencing; transcriptomics

Abstract Changes in the aged brain that occur with stroke, head injury or Alzheimer’s Disease Related Dementias (ADRD) result in chronic cognitive and motor deficits. Mesenchymal stem cells (MSCs) have recently received attention to reverse or slow neurodegenerative and injury-related changes in the aging brain as they suppress inflammation and facilitate tissue repair and remyelination. We have now completed studies as part of R21- NS102991 and R56-NS112207 that have shown that MSC-derived extracellular vesicles (EVs), the active product of MSCs, accelerate and enhance recovery of motor function following cortical injury in aged female monkeys. EV-treated monkeys exhibited full recovery by 3-5 weeks post-injury and untreated monkeys reached a plateau in recovery by 8-12 weeks post-injury. At a chronic recovery stage (16-weeks post-injury, R21- NS102991), MSC-EVs reduced injury-induced microglial neuroinflammation, neuronal excitotoxicity, synapse loss, oligodendrocyte damage and myelination deficits. However, the precise nature of how MSC-EVs act within both acute or chronic stages of recovery after injury remains unknown and is important to decipher in order to fine tune the efficacious use of MSC-EVs for age-related injury and neurodegenerative diseases. Our preliminary analysis of injury- and MSC-EV-associated changes in blood and brain tissue collected at an earlier stage of recovery (6 weeks post-injury, R56-NS112207) showed early treatment-related differences in microglial phenotypes and neuronal excitability that are distinct from treatment-related differences observed at 16-week post-injury. Therefore, we now propose to continue and expand these studies to combine in vivo behavioral, MRI and proteomic CSF and blood biomarker analyses with comprehensive proteomic and single-cell transcriptomic profiling and physiological and histological assessments of brain tissue harvested at distinct time points during recovery (3, 6, 9 weeks post- injury) from male and females aged monkeys. Furthermore, using advance human- derived in-vitro models, we will elucidate specific encapsulated miRNAs or/and proteins of MSC-EVs that ameliorate injury-related inflammatory and oxidative responses, and facilitate recovery and neuroprotection. We hypothesize that MSC-EVs contain miRNA and protein signals that modulate the acute response to injury and mitigate immediate damage, leading to reduced secondary chronic inflammation and degeneration, promoting a restorative microenvironment that will facilitate neural plasticity later in recovery. These studies will elucidate the temporal progression, sex-dependent dynamics and cell-specific molecular mechanism(s) of action of MSC-EV mediated recovery, that will pave the way for its potential development as a therapeutic for age-related injury and neurodegenerative diseases in humans.

摘要 中风、头部损伤或阿尔茨海默病相关性痴呆（ADRD）引起的老年大脑变化 导致慢性认知和运动缺陷。间充质干细胞（MSCs）最近受到关注 逆转或减缓衰老大脑中神经退行性和损伤相关的变化， 炎症并促进组织修复和髓鞘再生。我们现在已经完成了R21的研究- NS 102991和R56-NS 112207的研究表明，MSC衍生的细胞外囊泡（EV），活性微囊泡， 骨髓间充质干细胞产物，加速和增强老年女性皮质损伤后运动功能的恢复 猴子EV治疗的猴子在受伤后3-5周表现出完全恢复，而未治疗的猴子在受伤后3-5周表现出完全恢复。 损伤后8-12周恢复平稳。在慢性恢复阶段（损伤后16周，R21- 24）， NS 102991），MSC-EV减少损伤诱导的小胶质细胞神经炎症，神经元兴奋性毒性，突触 缺失、少突胶质细胞损伤和髓鞘形成缺陷。然而，MSC-EV如何在 受伤后的急性或慢性恢复阶段仍然未知，重要的是要破译， 微调MSC-EV对年龄相关损伤和神经退行性疾病的有效使用。我们的初步 分析在早期阶段收集的血液和脑组织中的损伤和MSC-EV相关变化， 恢复期（损伤后6周，R56-NS 112207）显示小胶质细胞的早期治疗相关差异， 与第16周时观察到的给药相关差异不同的表型和神经元兴奋性 受伤后。因此，我们现在建议继续并扩展这些研究，将联合收割机体内行为、MRI 和蛋白质组学CSF和血液生物标志物分析，采用综合蛋白质组学和单细胞转录组学 在不同的时间点收集的脑组织的分析和生理学和组织学评估， 从雄性和雌性老年猴中恢复（损伤后3、6、9周）。此外，利用先进的人类- 衍生的体外模型，我们将阐明MSC-EV的特异性包裹的miRNA或/和蛋白质， 改善损伤相关的炎症和氧化反应，并促进恢复和神经保护。我们 假设MSC-EV含有调节对损伤急性反应的miRNA和蛋白质信号， 减轻直接损害，导致减少继发性慢性炎症和变性，促进 恢复性微环境，这将有助于神经可塑性恢复后期。这些研究将阐明 MSC-EV作用时间进展、性别依赖性动力学和细胞特异性分子机制 介导的恢复，这将为其作为年龄相关损伤治疗药物的潜在发展铺平道路 和神经退行性疾病。