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: 10666419
• 负责人: David Sung
• 金额: $ 4.64万
• 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/10666419
• 关键词: Ablation; Adult; Affect; Ameloblasts; Apoptosis; Behavior; Bromodeoxyuridine; California; Catalytic Domain; Cell Differentiation process; Cells; Cellular Assay; Chromatin; Chromatin Remodeling Factor; Complex; Dental; Dental Enamel; Dental Schools; Dental caries; Dentists; Dentition; Deposition; Development; Enhancers; Environment; Epithelial Cells; Epithelium; 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; Methylation; Modeling; Mus; Natural regeneration; Organ; Pattern; Polycomb; Population; Proliferating; Proteins; Public Health; Qualifying; Regulation; Repression; Research; 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; clinical training; 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.

项目总结/摘要 了解牙齿上皮祖细胞命运决定的机制将有助于奠定长期的 为干细胞生物学在人类牙列中的临床应用奠定基础。在不断生长的老鼠体内 切牙，循环祖细胞在稳态和损伤期间协调切牙上皮更新- 诱导再生祖细胞自我更新和分化的快速和精心协调的平衡 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。为了理解机制 细胞命运的决定，这项建议将研究如何polycomb抑制复合物2（PRC2），通过 组蛋白3上赖氨酸27的三甲基化抑制基因表达以驱动祖细胞的命运定型。 该提议将确立PRC2催化亚基Zeste同源物2增强子（EZH2）在切牙中的作用， 上皮细胞和其特定的染色质靶点在祖细胞分化。目标1将研究EZH2如何 有助于切牙上皮祖细胞的命运决定在稳态和损伤诱导 再生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， 例如分化、自我更新或凋亡的变化。目的2阐明EZH2的特异性靶点 在体内平衡和损伤诱导的再生过程中的祖细胞。切牙的染色质状态 将使用最先进的转座酶单细胞测定法测定两种条件下的上皮细胞 使用核酸酶的可重复染色质测序（scATACseq）和靶下切割和释放 (CUT& RUN）技术。这些分析将显示PRC2在切牙上皮中的靶点是否相似 与其他自我更新组织中的细胞相比。他们还将确定牙齿上皮组织特异性靶点， 促成成釉细胞和非成釉细胞的命运。 这项研究计划将与一项全面的培训计划一起进行， 申请人作为牙医科学家的职业生涯。培训包括由高素质的 临床医生-科学家赞助，以及通过参加研讨会，期刊俱乐部， 课程、实验室会议、会议等等。研究和培训将在哥伦比亚大学进行。 加州，旧金山弗朗西斯科，它提供了一个优秀的研究环境和一个优秀的牙科学校 临床培训。