Enhanced cellular therapy for neonatal stroke

项目总结 新生儿卒中是导致死亡和残疾的重要原因，由多种细胞死亡途径引起。 以及随着时间的延长而演变的功能障碍。对该角色的了解不足 血管生成在局部缺血-再灌注损伤中的反应，这是早期最常见的原因 卒中。血管生成和神经元再生在很近的地方发生，旁分泌因子支持 神经元与内皮细胞的相互作用对修复至关重要。调节这个神经血管生态位可能是一种 提高新生儿期缺血性脑损伤后预后的潜在靶点。基于细胞的疗法 已经成为治疗中枢神经系统疾病的一种有前途的疗法，尽管修复的机制一直是 有争议的。间充质干细胞(MSC)在血管形成中发挥作用，并分泌促血管生成作用。 因子，表达许多生长因子受体，并可能优先分化为能够 在受伤后建立新的血管。骨髓间充质干细胞治疗已被证明改善了组织学和功能 缺血性损伤后的结果，即使治疗被推迟了，但只能部分修复。正在修改 MSC在某种程度上增强了对神经血管单位的影响，可能提供额外的好处。 我们的总体目标是确定延迟细胞疗法的再生和修复机制。 通过关注受损大脑中的血管反应来治疗新生儿中风。在目标1中，我们将测试 MSC预先暴露于EPO将促进血管生成和血管重塑的假说 新生大鼠局灶性脑缺血再灌注损伤程度大于MSC或EPO单独治疗。在目标2中，我们将 通过使用一些技术，阐明这种改良的细胞疗法在多大程度上调节长期修复 以量化细胞命运、大体组织学和长期感觉运动和认知结果。最后，在目标3中，我们 将确定特定的下游信号通路在缺血后血管生成和修复中的作用。 再灌注损伤和细胞治疗。这将确定关键的、可修改的路径 在发育中的大脑和局灶性损伤后的血管生成，这可以进一步研究，以增强长期的 定期修理。我们的初步假设是，用促红细胞生成素预处理的骨髓间充质干细胞延迟治疗将促进 血管生长和重塑，增加神经再生，改善长期组织学和功能 新生儿中风后的预后。这将为早期大脑的常见原因提供一种晚期治疗选择 损伤，在那里诊断往往被延误。

PROJECT SUMMARY Neonatal stroke is an important cause of death and disability, resulting from multiple pathways of cell death and dysfunction that evolve over a prolonged period of time. There is insufficient knowledge regarding the role of angiogenesis in the response to focal ischemia-reperfusion injury that is the most common cause of early stroke. Angiogenesis and neuronal repopulation occur in close proximity, with paracrine factors supporting neuron-endothelial cell interactions that are critical for repair. Modulating this neurovascular niche may be a potential target for enhancing outcomes after ischemic brain injury in the newborn period. Cell-based therapies have emerged as a promising treatment for CNS disease, although the mechanism of repair has been controversial. Mesenchymal stem cells (MSC) play a role in vascular formation and secrete pro-angiogenic factors, express a number of growth factor receptors, and may preferentially differentiate into cells capable of building new blood vessels after injury. MSC treatment has been shown to improve histological and functional outcomes after ischemic injury, even when therapy was delayed, but it is only partially reparative. Modifying MSC in a way that enhances effects on the neurovascular unit may provide additional benefit. Our overall objective is to determine the mechanism of regeneration and repair with delayed cellular therapy for neonatal stroke by focusing on the vascular response in the injured brain. In Aim 1, we will test the hypothesis that MSC pre-exposed to EPO will enhance angiogenesis and vascular remodeling following neonatal focal ischemia-reperfusion injury in the rat more than MSC or EPO therapy alone. In Aim 2, we will clarify to what extent this modified cellular therapy modulates long-term repair by using a number of techniques to quantify cell fate, gross histology and long-term sensorimotor and cognitive outcomes. Finally, in Aim 3, we will define the roles of specific downstream signaling pathways on angiogenesis and repair following ischemia- reperfusion injury and cellular therapy. This will determine critical, modifiable pathways important for angiogenesis in the developing brain and following focal injury, which can be further studied to enhance long- term repair. Our primary hypothesis is that delayed treatment with MSC pretreated with EPO will promote vascular growth and remodeling, increase neurogenesis, and improve long-term histological and functional outcomes after neonatal stroke. This will provide a late treatment option for a common cause of early brain injury, where diagnosis is often delayed.