Defining post-transcriptional regulons in intestinal epithelial regeneration

定义肠上皮再生中的转录后调节子

基本信息

批准号：
10375460
负责人：
Kathryn Elizabeth Hamilton
金额：
$ 53.08万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10375460
关键词：
ATG3 gene Affect Apoptosis Attenuated Autophagocytosis Binding Binding Proteins Biological Assay Cell Count Cell Line Cells Cellular biology Chemicals Clinical Colon Carcinoma DNA Damage Data Development Disease Epithelial Epithelial Cells Exhibits Flow Cytometry Gastrointestinal Injury Gene Deletion Genes Genetic Genetic Transcription Goals Homeostasis Hydroxychloroquine IGF2 gene Immunofluorescence Immunologic Infection Inflammation Injury Intestines Kinetics Knock-out LGR5 gene Link Luciferases Mediating Messenger RNA Modeling Molecular Mus Natural regeneration Paneth Cells Pathway interactions Patients Post-Transcriptional Regulation Predisposition Process Proteins RNA Biochemistry RNA-Binding Proteins Radiation Radiation Tolerance Reactive Oxygen Species Regenerative capacity Regulation Regulon Reporter Ribosomes Role Stress Testing Tissues Toxin Transcript Translations Up-Regulation Vesicle Wild Type Mouse Work cancer cell cell injury chemotherapy colon cancer cell line epithelium regeneration experience functional group gastrointestinal epithelium improved inhibition of autophagy innovation intestinal epithelium intestinal injury irradiation mouse model new therapeutic target novel strategies novel therapeutic intervention novel therapeutics overexpression prevent radiation-induced injury radioresistant repaired response ribosome profiling self-renewal stem cell function stem cells

项目摘要

Radiation, inflammation, infection, and toxins can damage the integrity of the gastrointestinal epithelium. Mechanisms that initiate, facilitate, and subsequently attenuate regeneration are essential to prevent long-term damage or disease. Post-transcriptional regulons are groups of functionally-related mRNAs regulated primarily by RNA-binding proteins, which enable rapid, tissue-specific responses to cellular damage. This type of regulation is especially important in highly proliferative tissues such as the intestinal epithelium, where RNA-binding proteins may serve as “rheostats” for cellular and tissue adaptation. Our overarching goal is to identify novel strategies to enhance intestinal epithelial regeneration and improve therapies for patients, especially those who experience injury to the gastrointestinal epithelium as a result of radiation or chemotherapy. We discovered recently that epithelial regeneration is enhanced in mice with intestinal epithelial deletion of IMP1 (IGF2 mRNA-binding protein 1; also called IGF2BP1), in part via up-regulation of the autophagy pathway. Autophagy is a mechanism by which cells adapt and survive in response to stress. For example, recent studies in mouse models with autophagy gene deletion demonstrate increased susceptibility to irradiation-induced tissue damage. Defining the link between IMP1 and autophagy would contribute to an emerging paradigm of post-transcriptional regulons that serve as critical regulators of gastrointestinal epithelial regeneration. Aim 1 will examine the molecular mechanisms by which IMP1 regulates autophagy gene translation. We will use cell biology assays as well as ribosome-profiling in epithelial cells from mice with Imp1 deletion to define direct roles for IMP1 in autophagy (and other) gene translation. Aim 2 will evaluate how IMP1-mediated changes in autophagy contribute to stem cell dynamics at homeostasis using mouse models, autophagy assays, and enteroid-formation assays. Aim 3 will evaluate intestinal stem cell-specific roles of IMP1 in autophagy-mediated repair during irradiation-induced injury and subsequent regeneration. We will use mouse models of Imp1 deletion together with chemical or genetic deletion of autophagy to define how IMP1 normally functions during irradiation-induced injury and regeneration and whether this is autophagy-dependent. Successful completion of this proposal has the potential for both scientific and clinical impact and will provide a basis for improved or novel therapies for patients with radiation-induced (and potentially other) injury to the intestinal epithelium.

辐射，感染和毒素会损害胃肠上皮的完整性。启动，促进和随后衰减再生的机制对于预防长期损害或疾病至关重要。转录后法规是通过RNA结合蛋白调节的功能相关mRNA组的组，这可以对细胞损伤产生快速，组织特异性的反应。这种调节在高度增殖的组织（例如肠上皮）中尤其重要，其中RNA结合蛋白可以用作细胞和组织适应的“变性”。我们的总体目标是确定新的策略，以增强肠上皮再生并改善患者的疗法，尤其是那些因放射或化学疗法而导致胃肠道上皮损伤的患者。我们最近发现，在IMP1的肠上皮缺失（IGF2 mRNA结合蛋白1；也称为IGF2BP1）的小鼠中，上皮再生得到了增强，部分是通过自噬途径的上调。自噬是一种机制，细胞可响应压力而适应和生存。例如，在具有自噬基因缺失的小鼠模型中的最新研究证明了对辐射引起的组织损伤的敏感性。定义IMP1和自噬之间的联系将有助于转录后法规的新兴范式，这些法规是胃肠道上皮再生的关键调节剂。 AIM 1将检查IMP1调节自噬基因翻译的分子机制。我们将在具有IMP1缺失的小鼠的上皮细胞中使用细胞生物学测定以及核糖体促进，以定义IMP1在自噬（和其他）基因翻译中的直接作用。 AIM 2将使用小鼠模型，自噬测定和肠托体形成测定法评估IMP1介导的自噬的变化如何对稳态的干细胞动力学有效。 AIM 3将评估IMP1在辐射诱导的损伤和随后再生过程中IMP1在自噬介导的修复中的肠道特异性作用。我们将使用IMP1缺失的小鼠模型以及自噬的化学或遗传缺失来定义IMP1在辐射诱导的损伤和再生过程中通常的功能以及这是否依赖于自噬。该提案的成功完成具有科学和临床影响的潜力，并将为辐射诱导（以及其他）肠上皮损伤的患者改善或新颖的疗法提供基础。