The role of the RNA binding protein IMP1 in intercellular communication and necrotizing enterocolitis

项目总结 坏死性小肠结肠炎(NEC)是一种影响早产儿肠道的破坏性炎症性疾病。 婴儿。在我们对NEC的病理生理学的理解上有很大的差距，包括对这种情况通常没有治愈方法。 致命的疾病。这项提议旨在通过定义发展中的 NEC过程中肠组织表达mRNA结合蛋白IMP1。IMP1在肠道屏障中的作用 维持、免疫细胞激活、肠道干细胞、自噬和细胞外小泡(EVS) 都可能影响NEC的病因学；然而，IMP1在NEC中的作用尚不清楚。在这里，我将检验这一假设 RNA结合蛋白IMP1转录后促进直接细胞到细胞和间接囊泡介导的 肠道内的沟通影响屏障和免疫功能及NEC的发育。在目标1中，我们将 利用肠道上皮中Imp1有条件地缺失或过表达的遗传性小鼠模型 比较对NEC攻击的生理和分子反应，重点是肠屏障功能。 我们将进行RNA免疫沉淀(RIP)测序，以确定新生儿中新的Imp1结合靶点 肠道和NEC期间，验证患者来源的肠道模型中的关键靶点，并检查 单层培养系统。在目标2中，我们将确定Imp1在EV沟通和肠道中的作用 激活巨噬细胞以保护其免受NEC的侵袭。这将通过直接分析电动汽车货物和 肠道巨噬细胞。我们将研究EVS、免疫细胞和上皮之间的相互作用 小鼠和患者来源的肠样细胞，以确定屏障功能和NEC病因学的机制。研究 将阐明肠上皮细胞通讯转录后调控的新机制， 有可能对我们对NEC的理解做出重大贡献。 我的目标是成为一名成功的独立科学家和胃肠病学领域的领导者， 确定新的治疗靶点以改进NEC治疗并最终预防NEC的长期目标。 在这个指导职业发展奖期间，我将在指导下完善现有的技能并获得额外的技能 我的研究指导委员会的成员，王博士，古德，马克斯，斯科托琳，索格斯塔德和布罗迪。除了……之外 根据我的导师和合作者的建议，我将继续学习免疫学和转录后研究 法规以及职业发展技能研讨会，包括生物伦理、赠款/手稿写作、 领导力和实验室管理。

PROJECT SUMMARY Necrotizing enterocolitis (NEC) is a devastating inflammatory disease that affects the intestine of premature infants. There is major gap in our understanding of the pathophysiology of NEC, including no cure for this often deadly disease. This proposal aims to help fill the knowledge gap by defining the role of the developmentally expressed mRNA binding protein IMP1 in the intestine during NEC. IMP1 plays roles in intestinal barrier maintenance, immune cell activation, intestinal stem cells, autophagy, and extracellular vesicles (EVs), which could all effect NEC etiology; however, the role of IMP1 in NEC is not known. Herein I will test the hypothesis that the RNA binding protein IMP1 post-transcriptionally promotes direct cell-to-cell and indirect vesicle-mediated communication in the intestine to impact barrier and immune function and NEC development. In Aim 1, we will utilize genetic mouse models where Imp1 is conditionally deleted or overexpressed in the intestinal epithelium to compare physiological and molecular responses to NEC challenge with a focus on intestinal barrier function. We will perform RNA immunoprecipitation (RIP)-sequencing to identify novel Imp1 binding targets in the neonatal intestine and during NEC, validate key targets in patient-derived enteroid models, and examine mechanisms in monolayer culture systems. In Aim 2, we will identify the role of Imp1 in EV communication and intestinal macrophage activation to protect against NEC. This will be achieved through direct analysis of EV cargo and intestinal macrophages. We will examine interactions between EVs, immune cells, and the epithelium using murine and patient-derived enteroids to define mechanisms governing barrier function and NEC etiology. Studies of IMP1 will elucidate novel mechanisms of post-transcriptional regulation of intestinal epithelial communication, with the potential to contribute significantly to our understanding of NEC. My goal is to become a successful, independent scientist and a leader in the field of gastroenterology, with the long-term goals of identifying new therapeutic targets to improve NEC treatment and ultimately NEC prevention. During this mentored career development award, I will refine existing and gain additional skills with the guidance of my research mentoring committee, Drs. Wong, Good, Marks, Scottoline, Saugstad, and Brody. In addition to the advice from my mentors and collaborators, I will pursue coursework in immunology and post-transcriptional regulation, as well as seminars in career development skills including bioethics, grant/manuscript writing, leadership, and laboratory management.