Using single cell transcriptomic analysis to uncover genetic pathways for de novo generation of dental epithelial progenitors

项目摘要 有效利用器官特异性体干细胞修复受损组织或生物工程器官将 革命性的疾病治疗和减轻老化和创伤造成的许多问题。但 精确地获得体干细胞并高效地扩增它们仍然具有重大挑战性 用于临床应用。技术障碍在很大程度上是由于我们对基因的不完全理解。 在发育过程中控制祖细胞命运特化的调节，以及成体中的细胞可塑性。 牙齿提供了一个很好的测试案例，以进一步了解这些方面，并应用于临床，作为成年人 牙齿不能维持牙齿上皮干细胞，并且缺乏再生能力。老鼠的牙齿是 强大的模型系统，研究器官发生和成体干细胞为基础的再生，并适合 体内遗传研究和离体操作以研究祖细胞功能。利用这一 卓越的实验系统，并将其与尖端的单细胞转录组学分析相结合， 该提案将深入了解遗传程序和转录过程中的变化， 牙上皮祖细胞的形成。这些知识将使我们能够识别一个基因网络和关键的 监管机构需要指定牙齿的命运，并提供一个蓝图，以获得牙齿祖细胞， 沿着遗传路径的多能干细胞。与此同时，该项目将测试IRX 1/2的功能和利用 和雅普在通过口腔上皮分化诱导牙祖细胞形成中的作用， 成釉细胞脱分化。我们将比较诱导， 胚胎和成年祖细胞，以确定IRX 1/2和雅普是否可以激活牙齿遗传程序。 基于这些数据，我们还将能够解决干细胞生物学中的一个重要问题，即如何 胚胎和成体中的不同祖细胞类型在转录上彼此相似。的主要创新点 该项目源于基因组技术和小鼠遗传模型的整合，以了解 牙齿祖细胞和干细胞形成的遗传调控，这是研究不足。这样的知识将形成 未来研究和赠款申请的基础，并使发展原则驱动的方法， 牙齿生物工程

PROJECT SUMMARY Effective utilization of organ-specific somatic stem cells to repair injured tissues or to bioengineer organs will revolutionize disease treatment and relieve numerous problems caused by aging and trauma. However, it remains significantly challenging to derive somatic stem cells with precision and expand them with high efficiency for clinical applications. The technical hurdles are in large part due to our incomplete understanding of the genetic regulation that controls fate specification of progenitors during development, as well as cell plasticity in adults. Teeth provide an excellent test case to further understand these aspects and apply clinically, as adult human teeth do not maintain dental epithelial stem cells and lack the capability to regenerate. The mouse tooth is a powerful model system to study both organogenesis and adult stem cell-based regeneration, and amenable for both in vivo genetic studies and ex vivo manipulations to investigate progenitor cell functions. Leveraging this remarkable experimental system and combining it with cutting-edge single cell transcriptomic analysis, this proposal will deliver an in depth understanding of the genetic program and transcriptional changes during the formation of dental epithelial progenitors. This knowledge will allow us to identify a genetic network and critical regulators required to specify the dental fate and provide a blueprint to derive dental progenitors by differentiating pluripotent stem cells along a genetic path. In parallel, this project will test the function and utilization of IRX1/2 and YAP in inducing the formation of dental progenitors through differentiation of oral epithelium and dedifferentiation of ameloblasts respectively. We will compare single cell transcriptomes between induced, embryonic, and adult progenitors to determine whether IRX1/2 and YAP can activate a dental genetic program. Based on these data, we will also be able to address an important question in stem cell biology, which is how different progenitor types in embryos and adults resemble each other transcriptionally. The main innovation of the project arises from the integration of genomic techniques and mouse genetic models to understand the genetic regulation of dental progenitor and stem cell formation, which is understudied. Such knowledge will form the basis of future research and grant applications, and enable developmental principle-driven approaches to tooth bioengineering.