Cord Blood is Neuroprotective in a Rat Model of Stroke

描述(申请人提供)：脑血管疾病是美国发病率和死亡率的第三大原因。随着人口老龄化，开发有效的治疗方案将变得更加紧迫，不仅可以增加存活率，还可以减少残疾。在中风(大脑中动脉阻塞或MCAO)的大鼠模型中，我们发现静脉注射(Iv)从人脐血(HUCB)中提取的单个核组分可以增强运动功能。在这项提案中，我们将探索如何通过直接的神经保护和抗炎机制来管理这些细胞群，以诱导MCAO的恢复和减少MCAO引起的解剖损伤。在目标1中，我们将使用炎症、神经元死亡、神经存活和凋亡的标记物来表征静脉注射HUCB细胞如何在MCAO后24小时至7天内改变MCAO的潜在病理和炎症。在目标2中，我们将确定有助于诱导行为和解剖恢复的特定HUCB细胞亚群。这将使用荧光激活细胞分选(FACS)来浓缩T细胞、B细胞、单核细胞和“干细胞”部分，以便移植到中风大鼠体内。这些研究中的终点测量将包括一系列运动测试和脑梗塞体积的表现。目的3提出了一系列的体外研究，以检验HUCB细胞是否通过与神经细胞群体的直接相互作用来改变MCAO的炎症反应。使用神经元、星形胶质细胞、少突胶质细胞或小胶质细胞进行细胞培养分析，以确定直接相互作用和介导缺氧/复氧反应中生存、细胞因子表达和核因子-kappaB结合活性的分子信号。在目标4中，我们将使用微阵列技术来识别神经修复/存活基因，这些基因在HUCB处理神经元培养后上调。所有的微阵列实验将通过免疫印迹和免疫组织化学进行验证。本文提出的研究将增加我们对HUCB细胞移植诱导卒中康复的神经保护和抗炎机制的理解，并可能确定卒中的其他潜在靶点，以开发治疗方案。这项研究计划的最终目标是开发一种创新的基于HUCB细胞的疗法，使其成为神经退行性疾病和脑损伤移植的可行临床选择。

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.