Cord Blood is Neuroprotective in a Rat Model of Stroke

脐带血对中风大鼠模型具有神经保护作用

• 批准号: 7101421
• 负责人: ALISON E WILLING
• 金额: $ 32.06万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-15 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7101421
• 关键词: clinical research; cord blood; infarct; model; stroke

DESCRIPTION (provided by applicant): Cerebrovascular disease is the third leading cause of morbidity and mortality in the United States. As the population ages, it will be even more pressing to develop effective treatment options that can not only increase survival, but decrease disability. In a rat model of stroke (the middle cerebral artery occlusion or MCAO) we found that intravenously (iv) administering the mononuclear fraction from human umbilical cord blood (HUCB) enhances motor functions. In this proposal we will explore how the administration of this population of cells may induce recovery and decrease anatomical damage induced by MCAO through direct neuroprotective and anti-inflammatory mechanisms. In Aim 1 we will characterize how iv HUCB cells modify the underlying pathology and inflammation of the MCAO when administered 24 hours to 7 days after the MCAO using markers of inflammation, neuronal death, neurosurvival, and apoptosis. In Aim 2, we will identify the specific subpopulation of the HUCB cells that is instrumental in inducing behavioral and anatomical recovery. This will be accomplished using fluorescent activated cell sorting (FACS) to enrich T cell, B Cell, monocyte and "stem" cell fractions for transplantation into a stroked rat. Endpoint measures in these studies will include performance on a battery of motor tests and infarct volume. Aim 3 proposes a series of in vitro studies to examine whether the HUCB cells modify the inflammatory response to the MCAO through direct interactions with neural cell popultions. Cell culture assays using neuronal, astrocytic, oligodendrocytes or microglia will be conducted to identify direct interactions and the molecular signals that mediate survival, cytokine expression and NF-kappaB binding activity in response to hypoxia/reoxygenation. In Aim 4 we will use microarray technology to identify neural repair/survival genes that are up-regulated by HUCB treatment of neuronal cultures. All microarray experiments will be verified with western blots and immunohistochemistry. The studies proposed here will increase our understanding of the neuroprotective and anti-inflammatory mechanisms underlying the recovery from stroke induced by HUCB cell transplantation and may also identify other potential targets in stroke for development of treatment options. The ultimate goal of this research program is to develop an innovative HUCB cell based therapy into a viable clinical option for transplantation in neurodegenerative disease and brain injury.

描述（由申请人提供）：脑血管疾病是美国发病率和死亡率的第三主要原因。随着人口年龄的增长，开发有效的治疗方案将更加紧迫，不仅可以增加生存，而且可以减少残疾。在中风的大鼠模型（中大脑中动脉闭塞或MCAO）中，我们发现静脉注射（IV）从人脐带血（HUCB）施用单核分数可以增强运动功能。在该提案中，我们将探讨该细胞群的给药如何通过直接神经保护和抗炎机制引起MCAO引起的恢复并减少解剖学损害。在AIM 1中，我们将表征IV HUCB细胞在MCAO后24小时至7天使用炎症，神经元死亡，神经外生和凋亡后24小时至7天进行MCAO的潜在病理和炎症。在AIM 2中，我们将确定对诱导行为和解剖学恢复的HUCB细胞的特定亚群。这将使用荧光活化的细胞分选（FACS）来富集T细胞，B细胞，单核细胞和“ Stem”细胞级分，以将其移植到抚摸大鼠中。这些研究中的端点度量将包括在一系列运动测试和梗塞量的情况下进行性能。 AIM 3提出了一系列体外研究，以检查HUCB细胞是否通过直接与神经细胞p的相互作用来改变对MCAO的炎症反应。将使用神经元，星形细胞，少突胶质细胞或小胶质细胞进行细胞培养测定，以鉴定直接相互作用以及介导生存期，细胞因子表达和NF-kappab结合活性的分子信号，以响应低氧/二氧化作用。在AIM 4中，我们将使用微阵列技术来识别通过HUCB治疗神经元培养的神经修复/生存基因。所有微阵列实验将通过蛋白质印迹和免疫组织化学进行验证。此处提出的研究将增加我们对HUCB细胞移植引起的中风恢复的基础神经保护和抗炎机制的理解，还可能识别出治疗方案开发的其他潜在目标。该研究计划的最终目的是将基于HUCB细胞的创新疗法开发为神经退行性疾病和脑损伤移植的可行临床选择。