Molecular and metabolic signaling in necrotizing enterocolitis

坏死性小肠结肠炎的分子和代谢信号传导

• 批准号: 10581835
• 负责人: DAVID J HACKAM
• 金额: $ 25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581835
• 关键词: 10 year old; Affect; Age; Award; Brain; Brain scan; Cause of Death; Cells; Cessation of life; Color; Communities; Contract Services; Data; Development; Disease; Equipment; Funding; Gastrointestinal Diseases; Goals; Hour; Human; Image; Intestines; Laparotomy; Lipopolysaccharides; Longevity; Lung; Maintenance; Metabolic; Microscope; Molecular; Mus; Necrosis; Necrotizing Enterocolitis; Organoids; Parents; Pathogenesis; Patients; Photobleaching; Premature Infant; Productivity; Sales; Sepsis; Signal Transduction; System; TLR4 gene; Time; Tissues; Training; cost; feeding; gut inflammation; improved; instrument; intestinal epithelium; neonatal mice; novel; novel therapeutics; prevent; receptor

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.

目前的要求是一个尼康AX共聚焦显微镜，这是需要为了完成 父R35的目标。 目前R35提案的目标是了解坏死性胰腺炎的潜在发病机制。 小肠结肠炎（NEC）是早产儿胃肠道疾病死亡的主要原因， 为这种毁灭性的疾病开发新的治疗方法。NEC的典型患者是早产儿， 婴儿从轻度喂养不耐受迅速发展为全身性脓毒症，然后在24小时内死亡。 NEC不成比例地影响有色人种社区，一半的患者需要剖腹手术， 显示出片状的肠道炎症和坏死NEC没有特异性治疗，增加了 迫切需要为这种毁灭性疾病开发新的疗法。在小鼠和 在人类中，PI已经发现革兰氏阴性细菌脂多糖的受体，即Toll样 肠上皮细胞上的受体4（TLR 4）是NEC发展所必需的。我们现在将这些 通过确定该领域四个关键的未回答的问题进行研究：1。什么原因导致NEC，以及为什么NEC是一个 肠内斑片状病变2.如何预防NEC？3.我们能提前预测NEC吗？4.什么原因导致 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 Ti Eclipse）已经超过10年，已经达到了它的寿命，并受到频繁故障的困扰。 现有的共焦也远远超出了其允许的服务合同，并鉴于其年龄，更换零件 很难买到。这些限制在最初提出建议时是没有预料到的， Nikon AX并没有提供演示。我们逐渐认识到， 完成家长奖的关键。我们有机会演示新的尼康AX 在实验室中，并留下了深刻的印象，其额外的能力，其中包括[1]两倍的视野（FOV）， 其对于捕获活细胞和组织类器官以避免光漂白（对于 目标1-3），以及[2]明显更快的图像采集时间，这对大脑切片特别有用， 对目标4至关重要。这些增强的功能将允许更干净的数据，由于较少的光漂白， 由于捕获时间增加（对于整个新生小鼠脑，从6小时到30分钟），生产率提高 扫描）。我们将利用现有资金支付与维护和培训有关的持续费用， 本行政裁决不包括采购费用。我们的尼康销售团队表示， 周的交货和组装，我们将有一个系统安装，并在一周内全面运作 的交付。