Precision Therapy for Neonatal Brain Injury

项目摘要/摘要部分 新生儿脑损伤是导致死亡和残疾的重要原因，通过氧化应激途径， 炎症和兴奋性毒性导致的损害，在很长一段时间内发展。治疗方法有 在损伤的早期阶段，经典的靶向单个通路，但在稍后的 损伤反应可能是额外有效的。治疗性低温(TH)，同时作为标准护理 对于缺氧缺血性脑病(HIE)，仅为60%的婴儿提供保护。最重要的是 假说是大脑在运动后的新陈代谢状态在 体温过低应答者和无应答者。我们将利用质子和核磁共振识别TH后的代谢状态 超极化碳13光谱，然后研究细胞途径，以识别更精确和 个体化治疗方法。我们将使用出生后第9天的小鼠，并跟踪它们直到受伤 进化论。本提案中概述的研究反映了一种创新和系统的方法来研究 新生儿脑损伤，因为它们结合了先进的代谢成像技术(质子和 碳谱)和细胞信号研究，重点是HIF信号，这将提供信息和 由人类临床研究提供信息。利用遗传细胞方法，其中损伤的组成部分 在特定的细胞隔间中特别删除/中断的反应将使我们能够评估 体内神经血管生态位的益处。我们将广泛询问依赖于HIF的信号通路 损伤后使用CHIP-SEQ。结合起来，这些遗传工具将允许我们探索分子 体外和体内对HI的调节以更好地寻找更合适的分子靶点用于治疗 最需要它们的人。通过使用传统方法在细胞水平上调查对HI的反应 生化分析和全球水平的大脑使用磁共振波谱，我们的目标是建立之间的联系 可非侵入性检测的特定细胞变化和代谢变化。这种方法将 最终允许将研究结果转化为临床，并有可能改变对 病人。 我们将使用CRISPR/Cas9和Chip-Seq等体外技术来剖析重要的信号通路 如缺氧诱导因子(HIF)。当我们确定适当的目标时，我们也将使用体外方法。 并设计治疗方法来抵消缺陷修复。因此，定义非脑血管疾病的脑代谢特征 应答者将确定后续的目标新途径，并将导致改善结果，可能 永远不能仅仅通过低温来实现靶向治疗。了解级联效应如何 每个阶段的关键调节因素将导致更多的基本治疗方法。

Project Summary/Abstract Section Neonatal brain injury is an important cause of death and disability, with pathways of oxidant stress, inflammation, and excitotoxicity leading to damage that progress over a long period of time. Therapies have classically targeted individual pathways during early phases of injury, but targeting pathways later in the injury response may be additionally effective. Therapeutic hypothermia (TH), while being standard of care for hypoxic-ischemic encephalopathy (HIE), provides protection only in 60% of babies. The overarching hypothesis is that the metabolic state of the brain immediately after TH differs markedly between hypothermia responders and non-responders. We will identify the metabolic state after TH using proton and hyperpolarized carbon 13 spectroscopy and then study cellular pathways to identify more precise and individualized treatment approaches. We will use postnatal day 9 mice and follow them through injury evolution. The studies outlined in this proposal reflect an innovative and systematic approach to the study of HI brain injury in the newborn because they combine advanced metabolic imaging techniques (proton and carbon spectroscopy) and cell-signaling studies, focusing on HIF signaling, that will both inform and be informed by human clinical studies. Utilizing genetic cellular approaches in which components of the injury response are specifically deleted/disrupted in the specific cell compartments will allow us to evaluate the benefits of the neurovascular niche in vivo. We will broadly interrogate HIF-dependent signaling pathways following injury using ChIP-Seq. Together, these genetic tools will allow us to explore the molecular regulation of HI both in vitro and in vivo to better identify more appropriate molecular targets for therapy for the individual needing them the most. By investigating responses to HI at a cellular level using traditional biochemical assays and global level in the brain using MR spectroscopy, we aim to make a link between specific cellular changes and metabolic changes that can be detected non-invasively. This approach would eventually allow the findings to be translated into the clinic and potentially change the management of patients. We will use in vitro techniques such as CRISPr/Cas9 and Chip-seq to dissect important signaling pathways like hypoxia inducible factor (HIF). We will also use an invitro approach when we identify appropriate targets and design therapies to counteract deficient repair. Thus, defining the cerebral metabolic signature of non- responders will identify subsequent novel pathways to target, and will lead to improved outcomes that could never be achieved by only targeting pathways through hypothermia alone. Understanding how the cascade of injury responses occur and the key modulators during each phase will lead to more rationale therapies.