Transcriptional Regulation of the Intestinal Epithelium

肠上皮的转录调控

基本信息

批准号：
7952403
负责人：
MICHAEL P. VERZI
金额：
$ 14.36万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7952403
关键词：
Address Adult Affect Arts Automobile Driving Binding Binding Sites Biological Assay Biological Models Biology CDX2 gene Cardiovascular system Cell Differentiation process Cell Maintenance Cells Communities Complex Coupling DNA Dana-Farber Cancer Institute Data Development Disease Environment Epithelial Epithelial Cells Epithelium Equipment Etiology Faculty Functional disorder Future Gastrointestinal tract structure Gene Expression Gene Targeting Genes Genetic Genetic Models Genome Goals Grant Hospitals Immune system Indium Individual Inflammatory Intestinal Diseases Intestines Knock-out Laboratories Lead Maintenance Malignant Neoplasms Maps Mediating Medical Mentors Mitotic Modeling Molecular Mucous body substance Mus Natural regeneration Nutrient Pattern Phenotype Play Population Postdoctoral Fellow Proliferating Property Regulator Genes Research Resistance Resources Role Scientist Staging Stem cells Technology Testing Thick Tissues Toxin Training Training Programs Transcriptional Regulation Villus Whole Organism Work career chromatin immunoprecipitation college gastrointestinal gastrointestinal function homeodomain in vivo insight interest intestinal crypt intestinal epithelium loss of function medical schools new technology next generation nutrition pathogen progenitor programs public health relevance research study stem transcription factor transcription factor CDX2

项目摘要

DESCRIPTION (provided by applicant): Candidate and choice of Mentor: The purpose of this K01 application is to continue my training in transcriptional mechanisms of gastrointestinal development and disease with the long-term goal of a career pursuing and hopefully curing the molecular etiologies of GI disorders and diseases. My interest in transcriptional mechanisms began in college and became my focus during graduate training with Dr. Brian Black at UCSF where I studied the transcriptional regulation of MEF2C, a gene with important functions in cardiovascular development. These studies yielded important insights on how an individual gene is regulated. For my postdoctoral work I was interested in expanding these studies to understand not how only one gene was regulated, but to employ new technologies to uncover entire gene regulatory programs. I sought a mentor pursuing the molecular mechanisms of gastrointestinal function with a fantastic track-record in training young scientists with expertise in transcriptional regulation. Ramesh Shivdasani and his laboratory will provide an outstanding training environment to pursue the transcriptional regulatory mechanisms controlling intestinal epithelial functions. Project: The intestine undergoes continual cell turnover as cells differentiate from stem and progenitor cells to differentiated intestinal cells that quickly carry out their functions and then are shed into the lumen. Progenitor and differentiated cells have very different gene expression patterns but the factors controlling these expression patterns are incompletely described. The homeodomain transcription factor CDX2 has the properties of a master regulator of the intestine in that it can dominantly induce intestine-type epithelium in non-intestinal tissues. Because CDX2 is expressed and has known target genes in both progenitor and differentiated cells, it is believed to play a role in mediating gene expression changes that occur during differentiation. Yet despite such profound functions, the molecular details of CDX2 actions are incompletely defined - particularly during the course of differentiation. My proposal applies recent advances in technology (whole genome chromatin immunoprecipitation) to comprehensively describe the molecular details of CDX2 function in both intestinal progenitor cells and their differentiated descendants. Firstly, CDX2 binding sites will be mapped in the genomes of progenitor cells and differentiated cells to determine whether the binding sites change concurrent with cellular state. This work will allow us to define how CDX2 can mediate gene expression changes during differentiation. Secondly, we have preliminary evidence to suggest CDX2 differentially cooperates with specific transcription factors in progenitor and differentiated cells, GATA6 and HNF4(, respectively. Whether CDX2 partners with these transcription factors and how these multi-factor complexes are assembled on the DNA will be determined. Finally, a proper genetic model to determine the requirement for CDX2 in the adult intestine will be generated. An inducible knockout of CDX2 will be analyzed to test the hypothesis that this factor is required for intestinal differentiation in the homeostatic adult intestine. Together, these experiments will provide molecular insights into how an important transcription factor mediates intestinal differentiation. This work will hopefully open avenues for future studies as an independent scientist where I hope to define gene programs susceptible to insults that result in intestinal disorders and cancers. Training Environment. The Dana Farber Cancer Institute and the affiliated hospitals of Harvard Medical School together provide tremendous resources to execute this training plan. Several instructional programs have been established to help transition late-stage postdoctoral fellows into independent scientists. In addition to full access to all necessary state-of-the-art research equipment and facilities, the Harvard Medical Community will be an asset by providing access to outstanding faculty from which I have assembled a mentoring committee comprised of experts in gastrointestinal biology and transcriptional mechanisms that will oversee my training progress and council me in my pursuit of an independent research program. Together with a strong mentor, well-founded research program and training plan, I hope this proposal will provide a strong beginning to a career investigating the molecular underpinnings of GI disease. PUBLIC HEALTH RELEVANCE: The intestinal epithelium is a one-cell-thick layer that is permissive to nutrients while resistant to toxins and pathogens within the intestinal lumen. This critical epithelial layer is susceptible to inflammatory diseases and cancer making it imperative to understand the molecular framework upon which normal functions are built. The proposed training grant will focus on developing a career pursuing the underlying transcriptional mechanisms of intestinal epithelial cell maintenance.

描述（由申请人提供）：候选人和导师的选择：这种K01应用的目的是继续我在胃肠道发展和疾病的转录机制方面进行培训，其长期目标是追求并希望能够治愈GI疾病和疾病的分子病因。我对转录机制的兴趣始于大学，并在UCSF的Brian Black博士的研究生培训中成为我的重点，在那里我研究了MEF2C的转录调节，MEF2C是一个在心血管发育中具有重要功能的基因。这些研究对如何调节单个基因产生了重要的见解。对于我的博士后工作，我有兴趣扩大这些研究，以了解不仅是一个基因受管制的，而是采用新技术来揭示整个基因调节程序。我寻求一位追求胃肠道功能的分子机制的导师，并在培训具有转录调节方面具有专业知识的年轻科学家方面具有出色的田径记录。 Ramesh Shivdasani和他的实验室将提供出色的培训环境，以追求控制肠上皮功能的转录调节机制。项目：随着细胞从茎和祖细胞区分到分化的肠细胞，肠道经历了连续的细胞更新，这些细胞迅速执行其功能，然后将其脱落到管腔中。祖细胞和分化细胞具有非常不同的基因表达模式，但是控制这些表达模式的因素不完全描述。同源域转录因子CDX2具有肠道主调节剂的特性，因为它可以主要诱导非智能组织中的肠道上皮。由于CDX2是表达的，并且在祖细胞和分化细胞中均具有靶基因，因此据信它在介导分化过程中发生的基因表达变化中起作用。然而，尽管功能如此深刻，但CDX2作用的分子细节仍未完全定义，尤其是在分化过程中。我的建议应用了技术的最新进展（整个基因组染色质免疫沉淀），可以全面地描述肠祖细胞及其分化后代中CDX2功能的分子细节。首先，CDX2结合位点将映射在祖细胞和分化细胞的基因组中，以确定结合位点是否与细胞状态同时变化。这项工作将使我们能够定义CDX2如何介导基因表达在分化过程中的变化。其次，我们有初步的证据表明，CDX2与祖细胞和分化细胞中的特定转录因子（分别为GATA6和HNF4）有差异合作（分别是与这些转录因子的伙伴合作伙伴，以及这些多因素复合物是否合作，以及这些多因素复合物如何在DNA上组装在DNA上，以确定CDX的适当构图。将分析以下假设，即在稳态成人肠道中进行肠道分化需要，这些实验将提供分子的见解，以介绍重要的转录因子如何介导这项工作，希望这项工作能够为未来的科学家开放，以便将其作为一个独立的科学家识别依据。培训环境。达纳·法伯癌症研究所（Dana Farber Cancer Institute）和哈佛医学院的附属医院共同提供了巨大的资源来执行该培训计划。已经建立了几个教学计划，以帮助过渡后期的博士后研究员到独立科学家。除了完全访问所有必要的最先进的研究设备和设施外，哈佛医学界还可以通过提供杰出教师的访问权来成为一种资产，我组建了一个由胃肠道生物学和转录机制专家组成的指导委员会，这些委员会将监督我的培训进度和理事会，以寻求独立的研究计划。我希望该提案将与强大的导师，结合的研究计划和培训计划一起，为调查GI疾病的分子基础的职业提供强大的开始。公共卫生相关性：肠上皮是一层单细胞层，允许营养，同时耐肠腔内的毒素和病原体。这种关键的上皮层容易患炎症性疾病和癌症，因此必须了解建立正常功能的分子框架。拟议的培训补助金将着重于发展肠道上皮细胞维护的基本转录机制的职业。