The role of IMP1 mRNA binding protein in intestinal epithelial biology

IMP1 mRNA结合蛋白在肠上皮生物学中的作用

• 批准号: 8766807
• 负责人: Kathryn Elizabeth Hamilton
• 金额: $ 11.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-09 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8766807
• 关键词: Ablation; Actins; Adult; Advisory Committees; Affect; Anoikis; Apoptosis; Autophagocytosis; Award; Binding; Binding Proteins; Bioethics; Bioinformatics; Biology; Biometry; Birth; Cells; Cellular biology; Clinical; Coculture Techniques; Colorectal Cancer; Culture Techniques; Data; Defect; Development; Diagnosis; Digestive System Disorders; Diphtheria Toxin; Disease; Dwarfism; Embryonic Development; Epithelial; Epithelial Cells; Equilibrium; Extracellular Matrix; Floor; Foundations; Funding; Future; Gastroenterology; Genetic; Grant; Growth; Health; Homeostasis; Human; Immunoprecipitation; Inflammation; Injection of therapeutic agent; Injury; Intestinal Diseases; Intestines; Knock-out; Knockout Mice; Laboratories; Lead; Liver diseases; Malignant - descriptor; Malignant Neoplasms; Manuscripts; Mentors; Mesenchymal; Messenger RNA; Methods; Modeling; Molecular; Molecular Biology; Morphology; Mus; National Institute of Allergy and Infectious Disease; National Institute of Diabetes and Digestive and Kidney Diseases; Neonatal; Pathway interactions; Phenotype; Physiologic pulse; Play; Post-Transcriptional Regulation; Process; Publications; RNA; Regulation; Relative (related person); Reporter; Research; Role; Rosa; Sorting - Cell Movement; Stem cells; Tamoxifen; Testing; Tissues; Training; Transgenic Mice; United States National Institutes of Health; Validation; Villus; Work; Writing; Xenograft procedure; base; c-Myc Staining Method; cancer cell; career; career development; cell type; diphtheria toxin receptor; enhanced green fluorescent protein; injury and repair; innovation; intestinal crypt; intestinal epithelium; intestinal homeostasis; meetings; mouse model; novel; novel therapeutics; overexpression; postnatal; public health relevance; ranpirnase; repaired; research and development; response; response to injury; skills; symposium; tumor

DESCRIPTION (provided by applicant): The proposed training in this K01 application outlines an integrated plan of mentored research and career development activities as well as a specific strategy for my pathway to an independent research career in intestinal epithelial biology. This award will allow me to refine existing and gain additional skills with the guidance of my research mentor, Dr. Rustgi, as well as an interdisciplinary advisory committee of Drs. Wu (Chair), Kaestner, Lengner, and Lynch. In addition to the advice from my mentor and committee, I will pursue formal coursework in stem cell and RNA biology, and biostatistics/bioinformatics, as well as seminars in career development skills including bioethics, grant/manuscript writing and laboratory management. During this award period I will present my work, both formally and informally, at national research conferences (DDW, Experimental Biology, Keystone, Gordon Conferences), at floor lab meetings through the NIDDK P30 Center for Molecular Studies in Digestive and Liver Diseases and Division of Gastroenterology, and at meetings of the UPenn constituency of the NIDDK/NIAID U01 Intestinal Stem Cell Consortium. Through this proposal, I will explore the role of IMP1 mRNA binding protein as a modulator of intestinal homeostasis through the use of existing and novel mouse models, innovative ex vivo 3D cultures techniques, and RNA-immunoprecipitation-sequencing. We hypothesize that IMP1 is a critical modulator of normal intestinal homeostasis and response to injury, through cell-type specific effects in the epithelial and mesenchymal compartments of the intestine. In Aim 1, we will utilize mouse models to conditionally knockout Imp1 in specific tissue compartments (epithelial or mesenchymal), in order to elucidate the relative contribution of Imp1 in specific cll types during normal homeostasis and injury. In Aim 2, we have developed and will test a novel Imp1 reporter mouse, which will be used as a lineage-tracing model for Imp1+ cells and will also allow for specific and temporal ablation of Imp1+ cells. We will utilize the mouse models from Aim 1 for complimentary ex vivo enteroid cultures of intestinal crypts and niche components, and mechanistic studies identifying novel mRNA binding targets of Imp1. We anticipate that results from these studies will define Imp1's role in intestinal homeostasis (through direct actions on epithelial crypts or through modulation of peri-cryptal niche components), the direct consequence of Imp1 loss in the neonatal period and adulthood during normal intestinal homeostasis and challenge with injurious agents, and novel Imp1 mRNA binding targets that will provide the foundation for future studies with translational application in diagnosis and therapy of intestinal diseases. Studies of IMP1 will elucidate novel mechanisms of post-transcriptional regulation of intestinal epithelial growth, with potential to contribute significantly to our understanding of intestinal health and disease.

描述（由申请人提供）：本K 01申请中的拟议培训概述了指导研究和职业发展活动的综合计划，以及我通往肠上皮生物学独立研究职业生涯的具体策略。这个奖项将使我能够完善现有的和获得额外的技能与指导我的研究 导师Rustgi博士以及由Wu博士（主席）、Kaestner、Lengner和Lynch组成的跨学科咨询委员会。除了从我的导师和委员会的建议，我将追求在干细胞和RNA生物学，生物统计学/生物信息学，以及在职业发展技能，包括生物伦理学，赠款/手稿写作和实验室管理研讨会正式课程。在此获奖期间，我将在国家研究会议（DDW，实验生物学，Keystone，Gordon Conferences）上正式和非正式地介绍我的工作，通过NIDDK P30消化和肝脏疾病分子研究中心和胃肠病学部门的地板实验室会议，以及NIDDK/NIAID U 01肠道干细胞联盟的UPenn选区的会议。 通过这个提议，我将探索IMP 1 mRNA结合蛋白作为肠道稳态调节剂的作用，通过使用现有的和新的小鼠模型，创新的离体3D培养技术，和RNA免疫沉淀测序。我们假设IMP 1是正常肠道内稳态和对损伤的反应的关键调节剂，通过在肠的上皮和间充质隔室中的细胞类型特异性作用。在目标1中，我们将利用小鼠模型在特定组织隔室（上皮或间充质）中条件性敲除Imp 1，以阐明Imp 1在正常稳态和损伤期间在特定细胞类型中的相对贡献。在目标2中，我们已经开发并将测试一种新型的Imp 1报告小鼠，该小鼠将用作Imp 1+细胞的谱系追踪模型，并且还将允许对Imp 1+细胞进行特异性和暂时性消融。我们将利用来自目标1的小鼠模型进行肠隐窝和小生境组分的互补离体肠样培养，以及鉴定Imp 1的新型mRNA结合靶点的机制研究。我们预期这些研究的结果将确定Imp 1在肠道内稳态中的作用（通过对上皮隐窝的直接作用或通过调节隐窝周围的小生境成分），在新生儿期和成年期在正常肠内稳态和用有害物质攻击期间Imp 1损失的直接后果，以及新的Imp 1 mRNA结合靶点，为今后在肠道疾病诊断和治疗中的翻译应用研究奠定基础。IMP 1的研究将阐明肠上皮生长的转录后调节的新机制，有可能对我们理解肠道健康和疾病做出重大贡献。