Molecular and metabolic signaling in necrotizing enterocolitis

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没有具体的治疗方法， 迫切需要为这种毁灭性的疾病开发新的疗法。 促使我写这份建议书的旅程包括对NEC二十多年的研究， 包括一个统一的定理来解释NEC的发展，发现了几个 类已获得专利的抗NEC代理，最近，完成作为唯一的编辑器， 第一本专门介绍NEC的教科书的主编。这一记录给了我一个独特的Vantage位置， 确定该领域中最重要的未回答问题，以及创建计划以回答的经验 他们流行的观点认为，NEC是由细菌引起的过度炎症反应引起的。 早产儿肠道内的定植。在小鼠和人体研究中，我发现革兰氏阳性细胞的受体 肠上皮细胞上的阴性细菌脂多糖，即Toll样受体4（TLR 4）， 这是NEC发展所需要的。TLR 4活性在早产的人类和小鼠肠道衬里中较高 这反映了TLR 4在控制肠道发育中的作用。随后的 通过定殖细菌激活TLR 4导致炎症和NEC。这些观察形成了 我发现了一种新型的抗TLR 4分子，可以预防和治疗小鼠、仔猪和人类的NEC 离体组织。在目前的提案中，我们现在将解决NEC领域的四个关键知识差距 研究：1. NEC及其标志性斑片状肠坏死的原因是什么？为了回答这个问题，我们将使用 ssRNA-seq在小鼠和人类NEC组织和人类类器官中发现并随后修饰TLR 4- 导致NEC的依赖性遗传途径; 2.如何预防NEC？我们将评估饮食的作用 （母乳与配方奶粉）和免疫代谢对早产儿肠道炎症诱导的影响， 操纵饮食诱导的炎症途径以减少NEC; 3.我们能提前预测NEC吗？我们将 评估导致NEC的细胞和分子母体因素， 非侵入性地，并开发用于预测的机器学习算法; 4.是什么导致了长期 NEC对肺、脑和肠道的并发症？我们将在人源化类器官中进行发现阵列 平台以及小鼠和仔猪NEC模型，以确定发炎的肠道和终末器官之间的联系 损伤这些研究有望显著推进NEC领域，并为其提供特异性治疗。

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 treatment for NEC, which increases the urgency to develop novel therapies for this devastating disease. The journey which has led me to write this proposal includes over two decades of research into NEC, including the advancement of a unifying theorem to explain NEC development, the discovery of several classes of anti-NEC agents which have been patented, and most recently, the completion as sole Editor-in- Chief of the first textbook devoted to NEC. This track record gives me a unique vantage point in which to identify the most important unanswered questions in the field, and the experience to create a plan to answer them. Prevailing thinking suggests that NEC arises from an exuberant inflammatory response to bacterial colonization in the premature intestine. In mice and human studies, I discovered that the receptor for gram negative bacterial lipopolysaccharide, namely toll-like receptor 4 (TLR4), on the intestinal epithelium, is required for NEC development. TLR4 activity is higher in the premature human and mouse intestinal lining compared to the full-term intestine, which reflects TLR4's role in governing gut development. The subsequent activation of TLR4 by colonizing bacteria leads to inflammation and NEC. These observations formed the basis for my discovery of a novel class of anti-TLR4 molecules that prevent and treat NEC in mice, piglet and human tissue ex vivo. In the current proposal, we will now address four critical knowledge gaps in the field of NEC research: 1. What causes NEC and its hallmark patchy intestinal necrosis? To answer this, we will employ ssRNA-seq in mouse and human NEC tissue and human organoids to discover and then modify TLR4- dependent genetic pathways causing NEC; 2. How can we prevent NEC? We will assess the role of diet (breast milk vs. formula) and immunometabolism on the induction of inflammation in the preterm gut and then manipulate diet-induced inflammatory pathways to reduce NEC; 3. Can we predict NEC earlier? We will assess cellular and molecular maternal factors that contribute to NEC, image blood flow in the premature gut non-invasively, and develop machine learning algorithms for prediction; 4. What causes the long term complications of NEC on the lung, brain and gut? We will perform discovery arrays in humanized organoid platforms as well as mouse and piglet NEC models to identify the link between the inflamed gut and end organ injury. These studies promise to significantly advance the field of NEC, and to offer specific therapies for it.