Mechanisms of Intestinal Epithelial Differentiation

肠上皮分化机制

基本信息

批准号：
10407820
负责人：
Christopher Eugene Ellison
金额：
$ 4.13万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
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：
2020-07-16 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10407820
关键词：
3-Dimensional Adult Automobile Driving Binding Cancer Prognosis Cell Differentiation process Cells ChIP-seq Chromatin Chromatin Loop Chronic diarrhea Colon Carcinoma Complex Critical Pathways DNA-Directed RNA Polymerase Data Dietary Fats Differentiated Gene Digestion Digestive Physiology Disease Duodenum Enhancers Enterocytes Epithelial Equilibrium Event Failure Gene Expression Gene Expression Regulation Genes Genetic Models Genetic Transcription Genome Goals HNF4A gene Health Human Human Engineering Inflammatory Bowel Diseases Intestines Kidney Kinetics Knock-out Knowledge Lead MADH4 gene Malignant Neoplasms Malnutrition Measures Mediating Modeling Molecular Mus Nuclear Receptors Nucleic Acid Regulatory Sequences Organoids Population Predisposition Process Publications Regenerative Medicine Regulatory Element Research Signal Pathway Signal Transduction System Testing Therapeutic Tissues Transcription Elongation Transgenic Mice Transgenic Model Villus Work design differential expression epigenomics gene function genetic variant ileum in vivo innovation intestinal crypt intestinal epithelium new technology novel nutrient absorption overexpression parent grant promoter protein complex recruit therapeutic target transcription factor transcriptome sequencing virtual

项目摘要

Parent Grant Abstract Proper differentiation of the intestinal epithelium underlies nearly all aspects of intestinal health. For example, improper epithelial barrier function is a critical contributor to inflammatory bowel diseases, and improper differentiation leads to poor terminal digestion and nutrient absorption, and increases susceptibility to cancers. Transcription factors that promote differentiation in the intestinal epithelium are known, but a gap exists in understanding how these factors interact with the transcriptional machinery to drive intestinal differentiation. New technologies now make it feasible to measure chromatin looping events and RNA Polymerase kinetics in vivo. These processes are unexplored in the intestine and likely to have intestine-specific mechanisms. The long term goal of our work is to resolve and manipulate the mechanisms of intestinal differentiation; our central hypothesis is that unexplored transcriptional mechanisms are dynamically regulated as cells differentiate in the intestine. These mechanisms include: 1) enhancer-promoter chromatin looping, 2) RNA polymerase loading, and 3) transcriptional elongation. The proposal will also examine how the pro- differentiation BMP/SMAD4 signaling pathway intersects with transcription factors to drive differentiation. A new, compound knockout of both HNF4A and HNF4G transcription factors leads to severe disruption of intestinal differentiation. ChIP-seq and RNA-seq identify hundreds of genes that are activated upon villus differentiation and depend upon HNF4 for expression. This novel transgenic model provides a rigorous system to apply new technologies and define intestinal regulatory mechanisms driving differentiation. The rationale of this proposal is to discover intestine-specific transcriptional regulatory mechanisms that can be exploited for the gain of human health. The novel transgenic mouse will be leveraged to define regulatory mechanisms required to activate differentiation genes in vivo. Aim 1 will apply new technologies to tackle unexplored transcriptional mechanisms driving differentiation. Aim 2 will explore a new paradigm for intestinal differentiation through the coordinated actions of BMP signaling and HNF4 factors. We expect to identify new mechanisms through which differentiation of enterocytes is achieved. Aims 1 & 2 are significant in that they can help us understand the impacts of genetic variants at enhancers, identify novel co-regulators, and lead to therapeutics that alter the homeostatic balance of intestinal differentiation that are relevant for treatments of IBD, cancer, and in regenerative medicine. HNF4 and SMAD4 are both strongly implicated in human IBD and Colon Cancer. The proposed research is innovative in the use of novel transgenic models, state-of-the-art epigenomics approaches, and human organoid cultures. Innovative ideas include the new paradigm of differentiation control by concerted efforts of HNF4 and BMP signaling, and provide a new concept through which the field can move forward to find therapeutic opportunities to control epithelial differentiation.

家长补助金摘要肠上皮的正确分化几乎是肠道健康所有方面的基础。例如，上皮屏障功能不当是炎症性肠病的一个关键因素，分化导致最终消化和营养吸收不良，并增加患癌症的可能性。促进肠上皮分化的转录因子是已知的，但在了解这些因子如何与转录机制相互作用以驱动肠道分化。现在，新技术使得测量染色质循环事件和 RNA 聚合酶动力学成为可能体内。这些过程在肠道中尚未被探索，并且可能具有肠道特异性机制。我们工作的长期目标是解决和操纵肠道分化的机制；我们的中心假设是未探索的转录机制随着细胞的变化而动态调节在肠道内区分。这些机制包括：1) 增强子-启动子染色质环，2) RNA 聚合酶负载，以及 3) 转录延伸。该提案还将研究如何支持分化 BMP/SMAD4 信号通路与转录因子交叉驱动分化。 HNF4A 和 HNF4G 转录因子的新复合敲除导致严重破坏肠分化。 ChIP-seq 和 RNA-seq 鉴定了数百个在绒毛上激活的基因分化并依赖于 HNF4 的表达。这种新颖的转基因模型提供了严格的系统应用新技术并定义推动差异化的肠道调节机制。该提案的基本原理是发现肠道特异性转录调节机制，该机制可以为人类健康而加以利用。新型转基因小鼠将被用来定义监管激活体内分化基因所需的机制。目标1将应用新技术来解决未探索的驱动分化的转录机制。目标2将探索肠道的新范式通过 BMP 信号传导和 HNF4 因子的协调作用进行分化。我们期望发现新的实现肠上皮细胞分化的机制。目标 1 和 2 非常重要，因为它们可以帮助我们了解遗传变异对增强子的影响，识别新的共同调节剂，并导致改变肠道稳态平衡的治疗方法与 IBD、癌症和再生医学治疗相关的分化。 HNF4 和 SMAD4 两者都与人类 IBD 和结肠癌密切相关。拟议的研究在使用新颖的转基因模型、最先进的表观基因组学方面具有创新性方法和人类类器官培养。创新理念包括差异化控制的新范式通过HNF4和BMP信号的共同努力，并提供了一个新的概念，通过该概念可以推动该领域的发展展望寻找控制上皮分化的治疗机会。