Mechanisms of Intestinal Epithelial Differentiation

肠上皮分化机制

• 批准号: 10726640
• 负责人: Christopher Eugene Ellison
• 金额: $ 0.73万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 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/10726640
• 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; Epithelium; 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; 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; antagonist; design; differential expression; epigenomics; gene function; genetic variant; ileum; in vivo; innovation; intestinal crypt; intestinal epithelium; new technology; novel; nutrient absorption; overexpression; promoter; protein complex; recruit; therapeutic target; transcription factor; transcriptome sequencing; virtual

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.

肠上皮的适当分化是肠道健康几乎所有方面的基础。例如， 上皮屏障功能不当是导致炎症性肠道疾病的关键因素，并且不适当 分化导致末期消化和营养吸收不良，并增加对癌症的敏感性。 已知促进肠上皮分化的转录因子是已知的，但是存在差距 了解这些因素如何与转录机械相互作用以驱动肠道分化。 现在，新技术使测量染色质循环事件和RNA聚合酶动力学可行 体内。这些过程在肠道中没有探索，并且可能具有特异性机制。 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 中心假设是未开发的转录机制被动态调节为细胞 区分肠道。这些机制包括：1）增强剂促销染色质循环，2）RNA 聚合酶负载和3）转录伸长。该提案还将研究如何 分化BMP/SMAD4信号通路与转录因子相交以驱动分化。 HNF4A和HNF4G转录因子的新的复合敲除导致严重破坏 肠道分化。 chip-seq和RNA-seq识别数百个在绒毛上激活的基因 分化并依靠HNF4进行表达。这个新颖的转基因模型提供了严格的系统 应用新技术并定义肠道调节机制推动分化。 该提议的理由是发现可以特定于肠道的转录调节机制 被利用以获取人类健康。新型的转基因小鼠将被利用以定义调节 在体内激活分化基因所需的机制。 AIM 1将应用新技术来解决 未开发的转录机制推动了分化。 AIM 2将探索肠道的新范式 通过BMP信号传导和HNF4因子协调作用的分化。我们希望确定新的 实现肠细胞分化的机制。 目标1和2很重要，因为它们可以帮助我们了解增强子的遗传变异者的影响， 识别新颖的共同调节剂，并导致治疗剂改变肠道的体内平衡 与IBD，癌症和再生医学治疗相关的分化。 HNF4和SMAD4 两者都与人IBD和结肠癌有关。 拟议的研究在使用新型转基因模型，最先进的表观基因组学方面具有创新性 方法和人体器官培养。创新的想法包括新的差异控制范式 通过HNF4和BMP信号的共同努力，并提供了一个新概念，该概念可以通过该概念移动 转发寻找控制上皮分化的治疗机会。

项目成果

Dynamic RNA Polymerase II Recruitment Drives Differentiation of the Intestine under the direction of HNF4.

动态 RNA 聚合酶 II 募集在 HNF4 的指导下驱动肠道分化。

• DOI: 10.1101/2023.11.08.566322
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Vemuri,Kiranmayi;Kumar,Sneha;Chen,Lei;Verzi,MichaelP
• 通讯作者: Verzi,MichaelP

Multiple roles and regulatory mechanisms of the transcription factor HNF4 in the intestine.

• DOI: 10.3389/fendo.2023.1232569
• 发表时间: 2023
• 期刊: Frontiers in endocrinology
• 影响因子: 5.2

β-Catenin Drives Butyrophilin-like Molecule Loss and γδ T-cell Exclusion in Colon Cancer.

• DOI: 10.1158/2326-6066.cir-22-0644
• 发表时间: 2023-08-03
• 期刊: CANCER IMMUNOLOGY RESEARCH
• 影响因子: 10.1
• 作者: Suzuki, Toshiyasu;Kilbey, Anna;Casa-Rodriguez, Nuria;Lawlor, Amy;Georgakopoulou, Anastasia;Hayman, Hannah;Swe, Kyi Lai Yin;Nordin, Anna;Cantu, Claudio;Vantourout, Pierre;Ridgway, Rachel A.;Byrne, Ryan M.;Chen, Lei;Verzi, Michael P.;Gay, David M.;Vazquez, Ester Gil;Belnoue-Davis, Hayley L.;Gilroy, Kathryn;Kostner, Anne Helene;Kersten, Christian;Thuwajit, Chanitra;Andersen, Ditte K.;Wiesheu, Robert;Jandke, Anett;Blyth, Karen;Roseweir, Antonia K.;Leedham, Simon J.;Dunne, Philip D.;Edwards, Joanne;Hayday, Adrian;Sansom, Owen J.;Coffelt, Seth B.
• 通讯作者: Coffelt, Seth B.