Molecular and metabolic signaling in necrotizing enterocolitis

目前的申请是尼康AX共焦显微镜，这是完成 父级R35的目标。 目前R35提案的目标是了解坏死性疾病的潜在发病机制。 小肠结肠炎(NEC)是早产儿死于胃肠道疾病的主要原因，以及 为这种毁灭性疾病开发新的治疗方法。典型的NEC患者是早产儿 婴儿从轻度喂养不耐受迅速发展为全身性败血症，然后在24小时内死亡。 NEC对有色人种的影响不成比例，一半的患者将需要剖腹手术，这 可见斑片状的肠道炎症和坏死。对于NEC没有特效的治疗方法，增加了 迫切需要为这种毁灭性的疾病开发新的治疗方法。在对小鼠和 人类的PI已经发现革兰氏阴性细菌脂多糖的受体，即Toll样脂多糖 肠上皮细胞上的受体4(TLR4)是NEC发育所必需的。我们现在将延长这些 通过确定该领域的四个关键问题进行研究：1.是什么导致了NEC，为什么NEC是一种 肠道斑片状疾病？2.我们如何预防NEC？3.我们能更早地预测NEC吗？4.原因是什么 NEC对肺、脑和肠道的长期并发症？ 所要求的设备(尼康AX共聚焦显微镜)将取代我们现有的显微镜(尼康A1钛 已有10多年历史的月食)已经到了寿命的尽头，并经常出现故障。 现有的Conocus也远远超出了其允许的服务合同，并且考虑到它的年限，更换部件 都很难买到。在最初提议时没有预料到这些限制，并且 尼康AX目前还没有现成的演示。我们已经意识到，替代仪器是 对于完成家长奖来说是非常必要的。我们有机会演示新的尼康AX 在实验室中，并对其附加功能印象深刻，其中包括[1]两倍的视野(FOV)， 这对于捕获活细胞和组织类器官以避免光漂白特别有用(对于 AIMS 1-3)和[2]显著加快了图像采集时间，这对大脑切片和 这些增强的功能将由于减少光漂白而允许更干净的数据，以及 由于延长了捕获时间(整个新生小鼠大脑的捕获时间从6小时增加到30分钟)，提高了工作效率 扫描)。我们将使用现有资金来支付与维护和培训相关的持续成本，以及 本行政裁决未涵盖的采购费用。我们的尼康销售团队表示，需要4-6 几周内交付和组装，我们将在一周内安装一个系统并完全运行 送货上门。

The current request is for a Nikon AX Confocal microscope, which is needed in order to complete the aims of the parent R35. The goal of the current R35 proposal is to understand the underlying pathogenesis of necrotizing enterocolitis (NEC), the leading cause of death from gastrointestinal disease in premature infants, and to develop novel treatments for this devastating disease. The typical patient with NEC is a premature infant who rapidly progresses from mild feeding intolerance to systemic sepsis and then death within 24 hours. NEC disproportionately affects communities of color, and half of all patients will require laparotomy, which reveals patchy intestinal inflammation and necrosis. There is no specific therapy for NEC, increasing the urgency for the development of novel therapies for this devastating disease. In studies in both mice and humans, the PI has discovered that the receptor for gram negative bacterial lipopolysaccharide, namely toll-like receptor 4 (TLR4), on the intestinal epithelium, is required for NEC development. We will now extend these studies by identifying four key unanswered questions in the field: 1. What causes NEC, and why is NEC a patchy disease in the intestine? 2. How can we prevent NEC? 3. Can we predict NEC earlier? 4. What causes the long term complications of NEC on the lung, brain and gut? The requested equipment (Nikon AX Confocal microscope) will replace our existing microscope (Nikon A1 Ti Eclipse) which is over 10 years old, has reached the end of its lifespan, and is beset by frequent breakdowns. The existing confocal is also well beyond its allowable service contract, and given its age, replacement parts are very difficult to purchase. These limitations were not anticipated at the time of the initial proposal, and the Nikon AX was not readily available to demo. We have come to realize that a replacement instrument is critically needed for completion of the parent award. We have had the opportunity to demo the new Nikon AX in the lab, and were impressed by its additional capabilities, which include [1] twice the field of view (FOV), which is especially useful for the capture of live cells and tissue organoids to avoid photobleaching (critical for Aims 1-3), and [2] significantly faster image acquisition time, which is especially useful for brain sections and critical for Aim 4. These enhanced features will allow for cleaner data due to less photobleaching, and improved productivity due to increased capture time (from 6h to 30 min for a whole neonatal mouse brain scan). We will use existing funds to cover ongoing costs associated with maintenance and training, and purchase costs not covered by this administrative award. Our Nikon sales team indicates that it takes 4-6 weeks for delivery and assembly, and that we will have a system installed and fully operational within a week of delivery.