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

基本信息

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
项目类别：
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

项目摘要

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 K01 申请中拟议的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之路的具体策略。该奖项将使我能够在研究的指导下完善现有技能并获得额外的技能导师 Rustgi 博士，以及 Drs. 的跨学科咨询委员会。吴（主席）、Kaestner、Lengner 和 Lynch。除了导师和委员会的建议外，我还将学习干细胞和 RNA 生物学、生物统计学/生物信息学的正式课程，以及职业发展技能研讨会，包括生物伦理学、资助/手稿写作和实验室管理。在此奖励期间，我将在国家研究会议（DDW、实验生物学、Keystone、Gordon Conferences）、通过 NIDDK P30 消化和肝脏疾病分子研究中心和部门的实验室会议上正式和非正式地展示我的工作胃肠病学，以及 NIDDK/NIAID U01 肠道干细胞联盟宾夕法尼亚大学选区的会议。通过本提案，我将通过使用现有和新型小鼠模型、创新的离体 3D 培养技术和 RNA 免疫沉淀测序，探索 IMP1 mRNA 结合蛋白作为肠道稳态调节剂的作用。我们假设 IMP1 通过对肠道上皮和间质室的细胞类型特异性作用，是正常肠道稳态和损伤反应的关键调节剂。在目标 1 中，我们将利用小鼠模型有条件地敲除特定组织区室（上皮或间质）中的 Imp1，以阐明正常稳态和损伤期间特定 cll 类型中 Imp1 的相对贡献。在目标 2 中，我们开发了一种新型 Imp1 报告小鼠，并将对其进行测试，该小鼠将用作 Imp1+ 细胞的谱系追踪模型，并且还可以对 Imp1+ 细胞进行特异性和暂时的消融。我们将利用 Aim 1 的小鼠模型对肠隐窝和生态位成分进行免费的离体肠样培养，并进行机制研究以确定 Imp1 的新 mRNA 结合靶点。我们预计这些研究的结果将定义Imp1在肠道稳态中的作用（通过对上皮隐窝的直接作用或通过调节隐窝周围的微环境成分），这是新生儿期和成年期正常肠道稳态和挑战期间Imp1丢失的直接后果具有有害物质和新的 Imp1 mRNA 结合靶点，将为未来在肠道疾病的诊断和治疗中的转化应用研究奠定基础。 IMP1 的研究将阐明肠上皮生长转录后调节的新机制，有可能为我们对肠道健康和疾病的理解做出重大贡献。