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

RNA结合蛋白IMP1在细胞间通讯和坏死性小肠结肠炎中的作用

• 批准号: 10610972
• 负责人: Sarah Andres
• 金额: $ 15.92万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-18 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610972
• 关键词: Adherent Culture; Affect; Autophagocytosis; Bacteria; Binding; Binding Proteins; Bioethics; Biology; Cell Communication; Cell Maintenance; Cells; Communication; Development; Disease; Epithelium; Etiology; Exposure to; Foundations; Functional disorder; Funding; Gastroenterology; Genetic; Goals; Grant; Human; Hypersensitivity; IGFBP2 gene; Immune; Immune response; Immune system; Immunology; Immunoprecipitation; Infant; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Ingestion; Integrins; Intestines; K-Series Research Career Programs; Knock-out; Knowledge; Laboratories; Lamina Propria; Leadership; Leaky Gut; Life; Lipids; Macrophage; Macrophage Activation; Maintenance; Manuscripts; Maps; Mediating; Mentors; Messenger RNA; MicroRNAs; Modeling; Molecular; Mucous Membrane; Mus; Necrotizing Enterocolitis; Neonatal; Nutrient; Pathogenesis; Pathway interactions; Patients; Peptides; Physiological; Play; Population; Post-Transcriptional Regulation; Pregnancy; Premature Birth; Premature Infant; Prevention; Prevention strategy; Production; Proteins; RNA; RNA immunoprecipitation sequencing; RNA-Binding Proteins; Research; Role; Scientist; Small Intestines; System; Techniques; Testing; Transcript; United States National Institutes of Health; Vesicle; Work; Writing; career; career development; chemokine; cytokine; diagnostic strategy; dysbiosis; extracellular vesicles; gastrointestinal; gut microbiota; immune activation; immune cell infiltrate; immune function; improved; inhibition of autophagy; intercellular communication; intestinal barrier; intestinal epithelium; migration; mortality risk; mouse model; nano-string; neonate; new therapeutic target; novel; novel diagnostics; overexpression; polarized cell; postnatal; posttranscriptional; premature; response; skill acquisition; skills; stem cell division; stem cells; treatment strategy

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.

项目概要 坏死性小肠结肠炎 (NEC) 是一种破坏性炎症性疾病，影响早产儿的肠道 婴儿。我们对 NEC 病理生理学的理解存在重大差距，包括通常无法治愈该病 致命的疾病。该提案旨在通过定义发展中的角色来帮助填补知识空白 NEC 期间肠道中表达 mRNA 结合蛋白 IMP1。 IMP1 在肠道屏障中发挥作用 维持、免疫细胞激活、肠道干细胞、自噬和细胞外囊泡 (EV)， 都可能影响 NEC 病因；然而，IMP1 在 NEC 中的作用尚不清楚。这里我将检验假设 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 肠道内的通讯影响屏障和免疫功能以及 NEC 的发展。在目标 1 中，我们将 利用肠上皮中 Imp1 有条件删除或过度表达的遗传小鼠模型 比较对 NEC 挑战的生理和分子反应，重点关注肠道屏障功能。 我们将进行 RNA 免疫沉淀 (RIP) 测序，以鉴定新生儿中新的 Imp1 结合靶点 肠和 NEC 期间，验证患者来源的肠样模型中的关键目标，并检查 单层培养系统。在目标 2 中，我们将确定 Imp1 在 EV 通讯和肠道中的作用 巨噬细胞活化以预防 NEC。这将通过直接分析电动汽车货物和 肠道巨噬细胞。我们将使用以下方法检查 EV、免疫细胞和上皮细胞之间的相互作用 小鼠和患者来源的肠样蛋白来定义控制屏障功能和 NEC 病因的机制。研究 IMP1的研究将阐明肠上皮通讯转录后调节的新机制， 有可能为我们对 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善 NEC 治疗并最终预防 NEC 的长期目标。 在这个受指导的职业发展奖期间，我将在指导下完善现有技能并获得额外的技能 我的研究指导委员会的博士。黄、古德、马克斯、斯科托林、索格斯塔德和布罗迪。此外 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课程 监管以及职业发展技能研讨会，包括生物伦理学、资助/手稿写作、 领导力和实验室管理。