Combinatorial Neuroprotective Strategies for Preterm Brain Injury

项目总结 早产仍然是全球和美国死亡率和发病率的主要原因。幸运的是， 在过去的几十年里，死亡率已经下降，以至于高收入人群的存活率现在超过了90% 国家。不幸的是，极早产儿神经发育结果的相应改善 婴儿(EP，出生和怀孕28周)仍然难以捉摸，超过60%的幸存者至少发育了一个 残疾，如脑瘫、自闭症、ADHD或认知、听力或视觉障碍。没有 当前临床使用的针对早产儿的定向神经保护干预措施，推动了显著的临床需求 开发降低EP婴儿死亡率和长期发病率的治疗方法。评估有希望的事情 早产儿的治疗，我们已经开发了补充的体外和体内技术 发育中的雪貂。炎症致敏低氧缺血高氧雪貂的初步研究 早产儿脑损伤模型显示出与婴儿一致的损伤模式和行为变化 早产的早产的在培养的器官型雪貂脑片上，暴露在缺氧-葡萄糖剥夺(OGD)中，我们 也表现出区域依赖的损伤，类似于早产人类，以及区域和治疗- 与神经保护相关的依赖转录组变化。我们看到的事实是，依赖于地区 对治疗的反应表明，最佳的治疗方法将需要联合治疗 提供全球神经保护并改善长期神经发育结果。这是特别的 考虑到最近发表的PENUT试验，没有发现显著的神经保护作用，这是相关的 促红细胞生成素(EPO)单一疗法--最有希望的疗法之一--在EP婴儿中的应用。 器官型脑片可以提供一个平台来筛选组合疗法，包括它们之间的相互作用。 例如，我们在相当于EP的雪貂脑片上的初步数据显示，EPO与抗- 炎症性抗生素阿奇霉素在皮质下白质产生协同效益，该区域 尤其是在EP婴儿中风险较高，但这种组合并不能在所有脑区都受益。在基础上建设 这些发现，我们提出的研究的目标是(1)确定区域特异性和有效性 多种有前景的神经治疗药物的体外比较，(2)评价联合神经治疗药物以优化局部 和体外的全球神经保护，以及(3)开发优化神经保护的神经疗法的鸡尾酒 在活体内建立了EP脑损伤的雪貂模型。我们的主要假设是：(1)神经治疗学 在体外提供互补的区域特异性神经保护将增加体内的全球神经保护，以及 (2)与单一疗法相比，联合补充神经疗法将产生更大的 整个大脑的神经保护一直持续到青春期。这项提议产生的数据可能 支持在这一人群中进行临床试验，目前还没有特定的神经保护疗法可用。

PROJECT SUMMARY Preterm birth remains a major cause of mortality and morbidity globally and in the United States. Fortunately, over the past several decades, mortality has decreased such that survival is now over 90% in high income countries. Unfortunately, commensurate improvements in neurodevelopmental outcomes of extremely preterm infants (EP, born <28 weeks’ gestation) remain elusive, with more than 60% of survivors developing at least one disability such as cerebral palsy, autism, ADHD, or cognitive, hearing, or visual impairment. There are no targeted neuroprotective interventions for preterm infants in current clinical use, driving a significant clinical need to develop therapies that reduce the mortality and long-term morbidity seen in EP infants. To evaluate promising therapeutics in the preterm infant, we have developed complementary in vitro and in vivo techniques in the developing ferret. Our preliminary data in the inflammation sensitized hypoxic-ischemic-hyperoxic (HIH) ferret model of preterm brain injury shows injury patterns and behavioral changes consistent with those seen in infants born prematurely. In cultured organotypic ferret brain slices exposed to oxygen-glucose deprivation (OGD), we have also shown regionally dependent injury, similar to the preterm human, and regional and treatment- dependent transcriptome changes associated with neuroprotection. The fact that we see regionally dependent responses to therapy suggests that an optimal therapeutic approach will require combinatorial therapies to provide global neuroprotection and improve long-term neurodevelopmental outcomes. This is particularly relevant considering the recent publication of the PENUT trial, which found no significant neuroprotective effect of erythropoietin (Epo) monotherapy – one of the most promising therapies in the pipeline – in EP infants. Organotypic brain slices can provide a platform to screen combinatorial therapeutics including their interactions. For example, our preliminary data in EP-equivalent ferret brain slices shows Epo in combination with the anti- inflammatory antibiotic azithromycin results in synergistic benefit in the subcortical white matter, a region that is specifically at risk in EP infants, but this combination does not result in benefit in all brain regions. Building on these findings, the objectives of our proposed research are to (1) determine the regional specificity and efficacy of multiple promising neurotherapeutics in vitro, (2) evaluate combined neurotherapeutics to optimize regional and global neuroprotection in vitro, and (3) develop a cocktail of neurotherapeutics optimizing neuroprotection in vivo in a ferret model of EP brain injury. Our overarching hypotheses are that: (1) neurotherapeutics that provide complementary region-specific neuroprotection in vitro will increase global neuroprotection in vivo, and (2) that compared to monotherapy, combining complementary neurotherapeutics will result in greater neuroprotection across the entire brain that persists into adolescence. Data resulting from this proposal could support a clinical trial in this population for which no specific neuroprotective therapies are currently available.