Cord Blood is Neuroprotective in a Rat Model of Stroke

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，不仅可以提高生存率，还可以减少残疾。在大鼠中风模型（大脑中动脉闭塞或 MCAO）中，我们发现静脉注射（iv）人脐带血 (HUCB) 的单核部分可增强运动功能。在本提案中，我们将探讨如何通过直接的神经保护和抗炎机制来管理该细胞群，从而诱导恢复并减少 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、神经存活和细胞凋亡标记物来描述在 MCAO 后 24 小时至 7 天施用时，静脉注射 HUCB 细胞如何改变 MCAO 的潜在病理学和炎症。在目标 2 中，我们将确定有助于诱导行为和解剖学恢复的 HUCB 细胞的特定亚群。这将使用荧光激活细胞分选 (FACS) 来富集 T 细胞、B 细胞、单核细胞和“干”细胞部分以移植到中风大鼠体内来完成。这些研究中的终点测量将包括一系列运动测试的表现和梗塞体积。目标 3 提出了一系列体外研究，以检查 HUCB 细胞是否通过与神经细胞群的直接相互作用来改变对 MCAO 的炎症反应。将使用神经元、星形胶质细胞、少突胶质细胞或小胶质细胞进行细胞培养测定，以确定直接相互作用和介导响应缺氧/复氧的生存、细胞因子表达和 NF-κB 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元培养物的 HUCB 处理上调的神经修复/存活基因。所有微阵列实验都将通过蛋白质印迹和免疫组织化学进行验证。这里提出的研究将增加我们对 HUCB 细胞移植引起的中风恢复的神经保护和抗炎机制的理解，并且还可能确定中风的其他潜在靶点以开发治疗方案。该研究计划的最终目标是开发一种创新的 HUCB 细胞疗法，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.