Mechanisms underlying YY1 control of intestinal epithelial homeostasis

YY1控制肠上皮稳态的机制

• 批准号: 8689012
• 负责人: MICHAEL P. VERZI
• 金额: $ 7.75万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8689012
• 关键词: Ablation; Adult; Alkaline Phosphatase; Area; Behavior; Binding; Cell Count; Cell Cycle Arrest; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cells; ChIP-seq; Chromatin; Chromatin Structure; Complement; Contracts; Country; Data; Data Set; Differentiation Antigens; Disease; Duodenum; Embryo; Epithelial; Epithelial Cells; Equilibrium; Event; Funding Mechanisms; Gene Expression; Gene Expression Profiling; Gene Targeting; Genes; Genetic; Genome; Genomics; Goals; Health; Homeostasis; Intestinal Diseases; Intestines; Knock-out; Knowledge; Lead; Length; Location; Maps; Measures; Mediating; Medical; Mentored Research Scientist Development Award; Molecular; Mus; Mutant Strains Mice; National Institute of Diabetes and Digestive and Kidney Diseases; Population; Process; Proliferating; Public Health; Recovery; Regulation; Research; Research Personnel; Role; Staining method; Stains; Stem cells; Technology; Testing; Therapeutic Intervention; Tissues; Villus; Work; Wound Healing; YY1 Transcription Factor; absorption; base; crypt cell; embryonic stem cell; epigenomics; fight against; innovation; intestinal epithelium; intestinal homeostasis; jejunum; novel; novel therapeutics; progenitor; public health relevance; skeletal; transcription factor; tumor; tumorigenesis; villin

DESCRIPTION (provided by applicant): In the intestinal epithelium, the regulatory mechanisms that mediate the balancing act between proliferation and differentiation are incompletely defined, but are critical for maintaining normal homeostasis and its recovery from disease. Intestinal epithelial homoeostasis requires precise regulation: excess proliferation can lead to tumor formation, but insufficient proliferation would cause tissue collapse. The objective of this application is to bridge the gap in our understanding of intestinal homeostasis by investigating an unexpected transcription factor's essential role in this process. Preliminary studies show that inactivation of the transcription factor YY1 in the adult murine intestinal epithelium triggers massive imbalance of epithelial homeostasis: the progenitor population expands while the differentiated population contracts. Therefore, defining how YY1 controls epithelial turnover could provide a novel therapeutic entry point in restoring epithelial homeostasis in disease. This project's first Aim is to test 3 plausible, non-exclusive explanations for the increase in proliferating crypt cells in intestinal epithelia lacking YY1: 1) YY1 controls stem cell turnover, 2) YY1 controls crypt cell proliferation, or 3) YY1 mediates cell cycle arrest and terminal differentiation. Mouse genetic and histological approaches will measure epithelial cell populations and behaviors upon YY1 loss. Completion of this aim should define the specific process(es) by which YY1 maintains homeostatic balance. The second Aim of the project will define the transcriptional regulatory mechanisms underlying YY1's control of epithelial turnover. ChIP-seq will be employed to map YY1 interactions with the genome of isolated intestinal epithelial cell populations. Identification of YY1's genomic binding locations will allow us to identify its direct regulatory target genes, identify its regulatory partner transcription factors,and determine whether YY1 operates in active or repressed chromatin structure. These analyses will be integrated with gene expression analysis of the YY1 knockout, and complemented by chromatin ChIP-seq data collected from the current K01. The proposed research is innovative in that it is expected to reveal a novel, YY1-dependent regulatory mechanism governing the proliferation/differentiation decision by using cutting-edge epigenomic technologies. Completion of the proposed work will reveal new mechanisms underlying homeostatic cell turnover and should be of significant value to fields as diverse as wound healing and oncogenesis. Finally, as is intended for this R03 mechanism, funding this project should enhance the capability of the K01-awarded investigator to conduct research. Establishing new investigators to join the fight against intestinal disease is a priority for the NIDDK as part of a long-term goal to solve one of the Country's largest medical problems. The proposed research on the regulatory mechanisms of YY1 is perfectly suited for this purpose in that it can be completed in a short timeframe, yet i likely to open a new area of research on the mechanisms of intestinal epithelial renewal.

描述（由申请人提供）：在肠上皮中，介导增殖和分化之间介导平衡行为的调节机制未完全定义，但对于维持正常的体内平衡及其从疾病中恢复至关重要。肠上皮同性恋需要精确调节：过量增殖会导致肿瘤形成，但增殖不足会导致组织塌陷。该应用的目的是通过研究意外转录因子在此过程中的重要作用来弥合我们对肠内稳态的差距。初步研究表明，成年鼠肠上皮上皮的转录因子YY1失活触发上皮稳态的巨大失衡：祖先种群在差异化的同时扩大了人口合同。因此，定义YY1如何控制上皮周转率可以为恢复疾病中上皮稳态的新型治疗点提供。该项目的第一个目的是测试3个合理的，非排他性的解释 对于缺乏YY1：1）YY1控制干细胞转换的肠上皮中增殖的隐窝细胞的增加，2）YY1控制着隐窝细胞的增殖，或3）YY1介导细胞周期的停滞和末端分化。小鼠遗传和组织学方法将在YY1丧失后测量上皮细胞群体和行为。该目标的完成应定义YY1维持稳态平衡的特定过程。该项目的第二个目标将定义YY1对上皮周转的控制基础的转录调节机制。 CHIP-SEQ将用于将YY1相互作用与分离的肠上皮细胞群体的基因组绘制。鉴定YY1的基因组结合位置将使我们能够识别其直接调节靶基因，确定其调节伙伴转录因子，并确定YY1是否在活性或抑制的染色质结构中运行。这些分析将与YY1敲除的基因表达分析集成，并与从当前K01收集的染色质芯片seq数据相辅相成。拟议的研究具有创新性，因为它有望揭示出一种新型的YY1依赖性调节机制，该机制通过使用尖端的表观学技术来控制增殖/分化决策。拟议工作的完成将揭示稳态细胞转换的新机制，对于像伤口愈合和肿瘤发生一样多样化的田地应具有重要价值。最后，正如该R03机制的旨在，资助该项目应增强K01授予的研究人员进行研究的能力。建立新的调查人员参加反对肠道疾病的斗争是NIDDK的重中之重，这是解决该国最大的医疗问题之一的长期目标的一部分。关于YY1的调节机制的拟议研究非常适合此目的，因为它可以在短时间内完成，但我可能会开设有关肠上皮更新机制的新研究领域。