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

• 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
• 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

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.

项目摘要/摘要 了解牙齿上皮祖细胞命运决定的机制将有助于长期建立 干细胞生物学在人牙列中的临床应用的基础。在不断生长的鼠标中 门牙，循环祖细胞在体内骨骼和损伤期间协调切牙上皮的更新 诱导的再生。祖细胞自我更新和分化的迅速而良好的平衡 在产生搪瓷的成成木或非木质细胞中，可以支持牙齿的持续生长。 但是，控制这些快速脂肪决策的机制尚不清楚。了解机制 在细胞命运的决策中，该提案将通过 赖氨酸27在组蛋白3上的三甲基化反映了基因表达以驱动祖细胞的命运承诺。 该建议将确定切牙中Zeste同源2（EZH2）的PRC2催化亚基增强子的作用 祖细胞分化过程中上皮及其特异性染色质靶标。 AIM 1将检查EZH2如何 在稳态期间有助于切牙上皮祖细胞脂肪决策和受伤引起的 再生。这将通过确定祖细胞中EZH2的损失如何影响细胞脂肪的决策来实现这一目标， 例如改变分化，自我更新或凋亡。 AIM 2将阐明EZH2的特定目标 在体内稳态和损伤引起的再生过程中的祖细胞中。门牙的染色质状态 在两种情况下，上皮细胞将使用最新的单细胞测定法确定转座酶 靶标在靶标下的染色质测序（SCATACSEQ）和裂解，并使用核酸酶释放 （切割和运行）技术。这些分析将表明切牙上皮中的PRC2靶标是否相似 到其他自我更新组织中的人。他们还将确定牙齿上皮组织特异性的靶标 有助于成年细胞和非成年细胞命运。 该研究计划将与旨在开发的综合培训计划一起制定 申请人作为牙医科学家的职业。培训包括高度合格的结构化精神职位 临床科学家赞助商以及科学和技术培训，通过参加期刊俱乐部， 课程，实验室会议，会议等等。研究和培训将在大学进行 加利福尼亚州旧金山，既提供出色的研究环境，又提供出色的牙科学校 用于临床培训。