CRE MOUSE MODELS TO STUDY AMELOGENESIS

研究釉质形成的 CRE 小鼠模型

• 批准号: 10674714
• 负责人: Michael Lansdell Paine
• 金额: $ 41.25万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10674714
• 关键词: Age; Ameloblasts; Amelogenesis; Animal Model; Animals; Anions; Cell Line; Cells; Connexin 43; Crossbreeding; Cystic Fibrosis Transmembrane Conductance Regulator; Dental; Dental Enamel; Deposition; Development; Disadvantaged; Disease; Enamel Formation; Enamel Organ; Enterobacteria phage P1 Cre recombinase; Funding; Future; Gene Expression; Genes; Grant; Investments; Keratin; Knock-in; Knockout Mice; Knowledge; LoxP-flanked allele; Lung; Maryland; Modeling; Multiple Organ Failure; Mus; Mutant Strains Mice; National Institute of Dental and Craniofacial Research; Organ; Organoids; Pancreas; Pathology; Pathway interactions; Phase; Proteins; Publishing; Research; Research Personnel; Resources; Role; Salivary Glands; Skin; Technology; Testing; The Jackson Laboratory; Tissues; Tongue; Tooth structure; Trachea; ameloblastin; animal data; bronchial epithelium; conditional knockout; genomic locus; in vivo Model; mRNA Expression; mouse Cre recombinase; mouse model; mutant; novel; promoter; spatiotemporal; tool

PROJECT SUMMARY / ABSTRACT In June of 2017 a group of dental researchers met at the NIDCR to discuss the future of enamel research and concluded, in a summary statement, that in the field of enamel research, a lack of models to study enamel formation and disease has hampered recent progress. The group concluded that more appropriate ameloblast- like cell lines, investment in organoid and chip technology and novel animal models could be used to advance the field. With respect to animal models, I believe enamel researchers suffer from not having enamel organ- specific Cre recombinase mutant mouse. For the past 2 decades enamel researchers have used the Krt14-Cre (keratin 14-Cre recombinase) mutant to study enamel-specific activities by cross breeding with various loxP mouse lines. The significant disadvantage of using the Krt14-Cre mouse for enamel research is that Krt14 is expressed in multiple tissues including skin, bronchial epithelia, tongue, trachea, salivary glands, and many more organs, and because of this many of the developed loxP mouse lines are not appropriate to study amelogenesis. For example, mRNA expression levels of the anion exchanger protein (Slc4a2/AE2), or the cystic fibrosis transmembrane conductance regulator (Cftr), increases ~ 6-fold and 3-fold respectively in the enamel organ during maturation stage (compared to secretory stage), and beyond tooth formation both genes are widely expressed in lung and pancreas and are critical to their development. Both Slc4a2-null and Cftr-null mice have severe enamel pathologies. To use the Krt14-Cre mutant mouse to study the role of either AE2fl/fl or Cftrfl/fl would have significant limitations because these animals would predictably suffer from multiple organ failures at a young age. I propose to develop two animal models with a knockin of Cre recombinase into the ameloblastin (Ambn) and odontogenic ameloblast-associated (Odam) gene loci such that Cre expression is limited to secretory ameloblasts and maturation ameloblasts respectively. The UG3 stage of this grant will be devoted to the development of these two animal models; Ambn-Cre and Odam-Cre, and the UH3 phase will validate these two animals as unique in vivo models to study amelogenesis. At the completion of this project data from these animals will be published, and both lines deposited in an appropriate facility such as the Mutamt Mouse Resource and Research Centers (MMRRC).

项目概要/摘要 2017 年 6 月，一组牙科研究人员在 NIDCR 举行会议，讨论牙釉质研究的未来和 在总结声明中得出的结论是，在牙釉质研究领域，缺乏研究牙釉质的模型 形成和疾病阻碍了最近的进展。该小组得出的结论是，更合适的成釉细胞- 与细胞系一样，对类器官和芯片技术以及新型动物模型的投资可用于推进 领域。就动物模型而言，我相信牙釉质研究人员因没有牙釉质器官而苦恼—— 特异性Cre重组酶突变小鼠。在过去的 20 年里，牙釉质研究人员一直使用 Krt14-Cre （角蛋白 14-Cre 重组酶）突变体通过与各种 loxP 杂交育种来研究牙釉质特异性活性 鼠标线。使用 Krt14-Cre 小鼠进行牙釉质研究的显着缺点是 Krt14 在多种组织中表达，包括皮肤、支气管上皮、舌头、气管、唾液腺等 更多器官，因此许多开发的 loxP 小鼠品系不适合研究 釉质发生。例如，阴离子交换蛋白 (Slc4a2/AE2) 的 mRNA 表达水平，或 囊性纤维化跨膜电导调节因子 (Cftr) 分别增加约 6 倍和 3 倍 牙釉质器官在成熟阶段（与分泌阶段相比），以及牙齿形成之后两个基因 在肺和胰腺中广泛表达，对其发育至关重要。 Slc4a2-null 和 Cftr-null 小鼠有严重的牙釉质病变。使用 Krt14-Cre 突变小鼠研究 AE2fl/fl 或 Cftrfl/fl 将具有显着的局限性，因为这些动物预计会遭受多器官疾病的折磨 年轻时的失败。我建议开发两种动物模型，将 Cre 重组酶敲入 成釉细胞蛋白 (Ambn) 和牙源性成釉细胞相关 (Odam) 基因位点，使得 Cre 表达为 分别限于分泌性成釉细胞和成熟成釉细胞。这笔补助金的 UG3 阶段将是 致力于这两种动物模型的开发； Ambn-Cre 和 Odam-Cre，以及 UH3 阶段将 验证这两种动物作为研究釉质形成的独特体内模型。在这个项目完成时 来自这些动物的数据将被公布，并且这两个品系都存放在适当的设施中，例如 突变小鼠资源和研究中心 (MMRRC)。