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: 9324209
• 负责人: Kathryn Elizabeth Hamilton
• 金额: $ 14.67万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 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/9324209
• 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; Histologic; Homeostasis; Human; Inflammation; Injection of therapeutic agent; Injury; Intestinal Diseases; Intestines; Knock-out; Knockout Mice; Laboratories; 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; RNA immunoprecipitation sequencing; Regulation; Reporter; Research; Role; Rosa; Stem cells; Tamoxifen; Testing; Tissues; Training; Transgenic Mice; Tumor Initiators; United States National Institutes of Health; Validation; Villus; Work; Writing; Xenograft procedure; base; c-myc Genes; cancer cell; career; career development; cell type; crosslinking and immunoprecipitation sequencing; 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; skill acquisition; skills; symposium; three dimensional cell culture

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)：此K01申请中建议的培训概述了指导研究和职业发展活动的综合计划，以及我在肠道上皮生物学独立研究职业生涯中的具体战略。这个奖项将使我能够在我的研究指导下改进现有的技能并获得额外的技能 导师，Rustgi博士，以及由Wu博士(主席)、Kaestner、Lengner和Lynch博士组成的跨学科咨询委员会。除了我的导师和委员会的建议外，我还将继续学习干细胞和RNA生物学、生物统计学/生物信息学的正式课程，以及职业发展技能的研讨会，包括生物伦理学、补助金/手稿写作和实验室管理。在此获奖期间，我将在国家研究会议(DDW、实验生物学、Keystone、Gordon会议)上，通过NIDDK P30消化和肝病分子研究中心和胃肠病学部在实验室会议上，以及在NIDDK/NIAID U01肠道干细胞联盟的宾夕法尼亚大学成员的会议上，正式和非正式地介绍我的工作。通过这项提议，我将通过使用现有的和新的小鼠模型、创新的体外3D培养技术和RNA免疫沉淀-测序来探索IMP1 mRNA结合蛋白作为肠道内稳态调节器的作用。我们假设IMP1是正常肠道内稳态和损伤反应的关键调节器，通过肠道上皮和间叶室的细胞类型特异性作用。在目标1中，我们将利用小鼠模型有条件地敲除特定组织间隔(上皮或间充质)中的Imp1，以阐明Imp1在正常内稳和损伤过程中特定CLL类型中的相对作用。在目标2中，我们已经开发并将测试一种新型的Imp1报告鼠，它将被用作Imp1+细胞的谱系追踪模型，并将允许对Imp1+细胞进行特异性和暂时的消融。我们将利用来自Aim 1的小鼠模型进行免费的肠隐窝和壁龛组件的体外肠样培养，并进行机制研究，以确定Imp1的新的mRNA结合靶点。我们预计，这些研究结果将确定Imp1‘S在肠道动态平衡中的作用(通过直接作用于上皮腺或通过调节隐窝周围生态位成分)、在新生儿和成年期正常肠道动态平衡期间Imp1丢失的直接后果以及新的Imp1mRNA结合靶点，这将为未来在肠道疾病的诊断和治疗中应用翻译应用的研究提供基础。对IMP1的研究将阐明肠上皮细胞生长的转录后调控的新机制，有可能对我们理解肠道健康和疾病做出重大贡献。