Dental Follicle: A Central Regulator of Tooth Root Formation and Regeneration

牙囊：牙根形成和再生的中央调节器

• 批准号: 10490624
• 负责人: Wanida Ono
• 金额: $ 37.49万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10490624
• 关键词: Ablation; Alleles; Alveolar; Apical; Area; Biology; CXC Chemokines; CXCL12 gene; Cell Therapy; Cells; Cementoblast; Characteristics; Defect; Dental; Dental Cementum; Dental Pulp; Dental Sac; Dental caries; Dentin; Epithelial; Epithelial Cells; Erinaceidae; Etiology; FOXF1 gene; Face; Foxes; Future; Gene Expression Profiling; Generations; Genes; Goals; Growth and Development function; In Situ; In Vitro; Intake; Knowledge; Ligands; Ligature; LoxP-flanked allele; Mastication; Mediating; Mesenchymal; Mesenchymal Stem Cells; Modeling; Molecular; Natural regeneration; Odontoblasts; Operative Surgical Procedures; Osteoblasts; Pathway interactions; Periodontal Diseases; Periodontal Ligament; Periodontitis; Periodontium; Physiological; Plant Roots; Play; Population; Population Heterogeneity; Prognosis; Prosthesis; Role; SHH gene; Signal Transduction; Stromal Cell-Derived Factor 1; Structure; Structure of periapical tissue; System; Testing; Tissues; Tooth Components; Tooth Diseases; Tooth Loss; Tooth regeneration; Tooth root structure; Tooth structure; Toxin; Transplantation; Validation; WNT Signaling Pathway; Work; alveolar bone; base; beta catenin; bone; cell type; comparative; experimental study; in vivo; insight; nutrition; parathyroid hormone-related protein; precursor cell; regenerative therapy; single-cell RNA sequencing; stem cells; tongue papilla; transcription factor; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT The tooth root is a critical component of the tooth, anchored to surrounding alveolar bones through the periodontal ligament (PDL). Appropriate formation of the root and its surrounding structure is essential not only for fundamental functions of the tooth in mastication for nutrition intake, but also for growth and development of the lower face. Prevalent dental diseases such as caries and periodontal diseases are the etiology for tooth loss, which can be treated only by prostheses lacking functional structures, leading to compromised long-term prognosis. An effective approach to regenerate a functional periodontal attachment apparatus is needed to achieve a breakthrough in dental regenerative therapies. During tooth root formation, the dental follicle (DF) provides precursor cells for cementoblasts, periodontal ligament (PDL) cells and alveolar cryptal bone osteoblasts to establish the functional periodontal attachment apparatus, the periodontium. Currently, how diverse populations of dental root mesenchymal progenitor cells work together and generate highly functional tooth roots remains unknown. In this proposal, we will define how concerted actions of distinct classes of dental root mesenchymal progenitor cells are essential for proper formation and regeneration of the tooth root accompanied by a functional periodontal attachment apparatus. In Aim 1, we will define how Hedgehog-Fox pathway regulates DF mesenchymal progenitor cells. We hypothesize that the Forkhead (Fox) transcription factors play a key role in regulating physiological functions of Hedgehog-responsive DF mesenchymal progenitor cells. We will first reveal the diversity of Hedgehog-responsive Gli1+ DF cells using single cell RNA- seq analyses, and determine their relationships with PTHrP+ DF cells and other precursor cell populations. Second, we will determine the function of Gli1+, PTHrP+ and Runx2+ cells in periodontium formation using inducible cell ablation experiments, and further test the function of Foxf1 and Foxf2 in periodontium formation using their floxed alleles. In Aim 2, we will identify Wnt-mediated roles of apical root mesenchymal progenitor cells in tooth root formation. We hypothesize that chemokine (C-X-C motif) ligand 12 (CXCL12)+ mesenchymal progenitor cells in the apical root area orchestrate formation of the tooth root and the apical periodontium in a canonical Wnt signaling-mediated manner. We will determine the relationship between CXCL12+ cells and stem cells for apical papilla (SCAP) using ex vivo culture system, and define molecular mechanisms underlying a Wnt-mediated cell fate choice of mesenchymal progenitor cells by a comparative RNA-seq analysis followed by in situ validation. In Aim 3, we will determine actions of dental root mesenchymal progenitor cells in periodontal regeneration. We hypothesize that distinct classes of dental root mesenchymal progenitor cells contribute to regeneration in a concerted manner. We will define how descendants of Gli1+, PTHrP+, Runx2+ and CXCL12+ progenitor cells contribute to periodontal regeneration after bone destruction, by utilizing a ligature-induced periodontitis model mimicking periodontal diseases, and a surgical periodontal defect model.

项目摘要/摘要 牙根是牙齿的关键成分，通过 牙周韧带（PDL）。根及其周围结构的适当形成不仅是必不可少的 牙齿在咀嚼营养摄入量中的基本功能，也用于生长和发展 下面。龋齿和牙周疾病等流行的牙齿疾病是牙齿的病因 损失，只能通过缺乏功能结构的假体来对待，导致长期损害 预后。需要一种有效的方法来再生功能牙周附着设备 在牙齿再生疗法中取得突破。在牙根形成期间，牙卵泡（DF） 提供胶合细胞，牙周韧带（PDL）细胞和肺泡隐性骨的前体细胞 成骨细胞以建立功能性牙周附着设备，即牙周。目前，如何 牙根间充质祖细胞的不同种群共同起作用并产生高功能 牙根仍然未知。在此提案中，我们将定义不同类别的共同行动 牙根间充质祖细胞对于正确形成和牙齿的再生至关重要 伴有功能性牙周附着设备。在AIM 1中，我们将定义刺猬福克斯 途径调节DF间充质祖细胞。我们假设叉子（fox）转录 因素在调节刺猬响应DF间充质的生理功能方面起关键作用 祖细胞。我们将首先揭示使用单细胞RNA- SEQ分析，并确定它们与PTHRP+ DF细胞和其他前体细胞群的关系。 其次，我们将使用牙周形成中Gli1+，PTHRP+和Runx2+细胞的功能 诱导细胞消融实验，并进一步测试FOXF1和FOXF2在牙周形成中的功能 使用它们的flox等位基因。在AIM 2中，我们将确定顶端根部充质祖细胞的Wnt介导的作用 牙根形成的细胞。我们假设趋化因子（C-X-C基序）配体12（CXCL12）+间质 根尖根区域中的祖细胞编排牙根的形成和顶端牙周牙周牙周 典型的Wnt信号介导的方式。我们将确定CXCL12+细胞和 使用离体培养系统的顶乳头（SCAP）的干细胞，并定义了分子机制 Wnt介导的细胞命运选择间充质祖细胞通过比较RNA-Seq分析随后 通过原位验证。在AIM 3中，我们将确定牙根间充质祖细胞在 牙周再生。我们假设不同类别的牙根间充质祖细胞 以一致的方式促进再生。我们将定义Gli1+，PTHRP+，Runx2+的后代如何 和CXCL12+祖细胞细胞通过利用A 结扎诱导的牙周炎模型模仿牙周疾病和手术牙周缺陷模型。