Chromatin modifier Polycomb Repressive Complex 2 as a regulator of dental epithelial progenitor cells

染色质修饰剂 Polycomb Repressive Complex 2 作为牙上皮祖细胞的调节剂

基本信息

批准号：
10535905
负责人：
David Sung
金额：
$ 4.42万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10535905
关键词：
Adult Affect Ameloblasts Apoptosis Behavior Bromodeoxyuridine California Catalytic Domain Cell Differentiation process Cells Cellular Assay Chromatin Chromatin Remodeling Factor Clinical Complex Dental Dental Enamel Dental Schools Dental caries Dentists Dentition Deposition Development Enhancers Environment Epithelial Epithelial Cells Equilibrium Fluorescence-Activated Cell Sorting Fluorouracil Gene Expression Gene Expression Regulation Genes Genetic Transcription Growth Histones Homeostasis Homologous Gene Human Immunofluorescence Immunologic Incisor Injury Journals Kinetics Label Laboratories Lysine Mediating Mentorship Modeling Mus Natural regeneration Organ Pattern Polycomb Population Proteins Public Health Regulation Repression Research Research Training Role San Francisco Scientist Stains Structure System TdT-Mediated dUTP Nick End Labeling Assay Technology Testing Tissues Tooth Loss Tooth regeneration Tooth structure Training Transposase Universities Wild Type Mouse career cell growth regulation chromatin modification clinical application design epithelial stem cell gene repression meetings mutant nuclease preservation progenitor promoter protein expression repaired self-renewal stem cell biology stem cells symposium

项目摘要

Project Summary/Abstract Understanding the mechanisms of dental epithelial progenitor cell fate decisions will help to lay the long-term groundwork for clinical applications of stem cell biology in human dentition. In the continuously growing mouse incisor, cycling progenitor cells orchestrate incisor epithelium renewal during both homeostasis and injury- induced regeneration. The swift and well-orchestrated balance of progenitor cell self-renewal and differentiation into either enamel-producing ameloblasts or non-ameloblasts supports the continuous growth of the tooth. However, the mechanisms that control these rapid fate decisions remain unclear. To understand mechanisms of cell fate decisions, this proposal will examine how Polycomb Repressive Complex 2 (PRC2), through trimethylation of lysine 27 on Histone 3, represses gene expression to drive fate commitment of progenitor cells. This proposal will establish the role of PRC2 catalytic subunit Enhancer of Zeste Homolog 2 (EZH2) in the incisor epithelium and its specific chromatin targets during progenitor differentiation. Aim 1 will examine how EZH2 contributes to incisor epithelial progenitor cell fate decisions during homeostasis and injury-induced regeneration. This will be achieved by determining how loss of Ezh2 in progenitor cells affects cell fate decisions, such as changes to differentiation, self-renewal or apoptosis. Aim 2 will elucidate the specific targets of EZH2 in progenitor cells during homeostasis and injury-induced regeneration. Chromatin states of incisor epithelial cells during both conditions will be determined using state-of-the-art single-cell Assay for Transposase Accessible Chromatin sequencing (scATACseq) and Cleavage Under Targets and Release Using Nuclease (CUT&RUN) technologies. These analyses will show whether PRC2 targets in the incisor epithelium are similar to those in other self-renewing tissues. They will also identify the dental epithelial tissue-specific targets that contribute to ameloblast and non-ameloblast fates. This research plan will be conducted in conjunction with a comprehensive training plan designed to develop the applicant’s career as a dentist-scientist. The training includes structured mentorship from a highly qualified clinician-scientist sponsor, as well as scientific and technical training through attending seminars, journal clubs, classes, laboratory meetings, conferences, and more. Research and training will take place at the University of California, San Francisco, which offers both an outstanding research environment and an excellent dental school for clinical training.

项目概要/摘要了解牙上皮祖细胞命运决定的机制将有助于奠定长期的基础为干细胞生物学在人类牙列中的临床应用奠定了基础。门牙，循环祖细胞在稳态和损伤期间协调门牙上皮更新诱导再生祖细胞自我更新和分化的快速且精心协调的平衡。进入产生釉质的成釉细胞或非成釉细胞，支持牙齿的持续生长。然而，控制这些快速命运决定的机制仍不清楚。细胞命运决定方面，该提案将研究 Polycomb Repressive Complex 2 (PRC2) 如何通过组蛋白 3 上的赖氨酸 27 发生三甲基化，抑制基因表达以驱动祖细胞的命运决定。该提案将确定 PRC2 催化亚基增强剂 Zeste 同源物 2 (EZH2) 在门牙中的作用目标 1 将研究 EZH2 在祖细胞分化过程中的上皮及其特定染色质靶点。有助于稳态和损伤诱导期间切牙上皮祖细胞的命运决定这将通过确定祖细胞中 Ezh2 的缺失如何影响细胞命运决定来实现， Aim 2 将阐明 EZH2 的具体靶标，例如分化、自我更新或细胞凋亡的变化。在门牙稳态和损伤诱导的染色质状态期间的祖细胞中。两种情况下的上皮细胞将使用最先进的单细胞转座酶测定来确定可访问染色质测序 (scATACseq) 以及使用核酸酶进行目标切割和释放 (CUT&RUN) 技术将显示切牙上皮中的 PRC2 目标是否相似。他们还将确定牙上皮组织的特异性靶标。有助于成釉细胞和非成釉细胞的命运。该研究计划将与旨在开发申请人作为牙医科学家的职业生涯包括来自高素质的结构化指导。临床医生-科学家赞助商，以及通过参加研讨会、期刊俱乐部进行科学和技术培训，课程、实验室会议、研究和培训将在大学举行。加利福尼亚州旧金山，提供出色的研究环境和优秀的牙科学校用于临床培训。