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

项目摘要 坏死性小肠结肠炎（NEC）是一种破坏性的炎症性疾病， 婴儿。我们对NEC的病理生理学的理解还有很大的差距，包括没有治愈的方法。 致命的疾病这项建议旨在通过界定发展中国家的作用， 在NEC期间肠中表达mRNA结合蛋白IMP1。IMP1在肠屏障中的作用 维持，免疫细胞活化，肠干细胞，自噬和细胞外囊泡（EV）， 可能都影响NEC病因;然而，IMP 1在NEC中的作用尚不清楚。在这里，我将测试假设 RNA结合蛋白IMP1在转录后促进直接的细胞间和间接的囊泡介导的 在肠道中的沟通，影响屏障和免疫功能和NEC的发展。在目标1中，我们 利用遗传小鼠模型，其中Imp1在肠上皮中条件性缺失或过表达 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。 肠和NEC期间，验证患者来源的肠模型中的关键靶标，并检查 单层培养系统。在目标2中，我们将确定Imp1在EV通信和肠道免疫中的作用。 巨噬细胞活化以保护免受NEC。这将通过对电动汽车货物的直接分析来实现， 肠道巨噬细胞我们将研究EV，免疫细胞和上皮细胞之间的相互作用， 小鼠和患者来源的类肠上皮细胞，以确定控制屏障功能和NEC病因的机制。研究 IMP1的表达将阐明肠上皮通讯的转录后调节的新机制， 有可能大大有助于我们对NEC的理解。 我的目标是成为一名成功的，独立的科学家和胃肠病学领域的领导者， 确定新的治疗靶点以改善NEC治疗并最终预防NEC的长期目标。 在这个指导职业发展奖，我将完善现有的，并获得额外的技能与指导 我的研究指导委员会的成员，王博士，古德，马克斯，斯科托林，索格斯塔德和布罗迪。除了 从我的导师和合作者的意见，我将追求在免疫学和转录后的课程 法规，以及职业发展技能研讨会，包括生物伦理学，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.